Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.018
Xu Jizhong, Gui-xian Wang, W. Yuan, Quanbo Zhou, Shengyun Hu
Objective To observe the effect of microRNA (miRNA, miR)-182 on the proliferation and migration of colorectal cancer cells by targeting special AT-rich sequence-binding protein 2 (SATB2) gene, and to explore its effect on SATB2/icotinamide adenine dinucleotide phosphate oxidase 4 (nox4) pathway. Methods HT-29 cells in logarithmic phase were transfected with Si-miR-182 and Control-si resectively, by liposome transfection, serving as Si-miR-182 group and N-miR-182 group, respectively. The expression of miR-182 gene, the cell proliferation and migration, the mRNA and protein expression of SATB2, nox4, E-cadherin and vimentin were examined. Results The relative expression of miR-182 gene in Si-miR-182 group (0.35±0.05) was lower than that in control group (1.24±0.26) and N-miR-182 group (1.20±0.25) (t=7.517, 7.455, P<0.05). The A values of methyl thiazol tetrazolium (MTT) test in Si-miR-182 group were lower than those in control group and N-miR-182 group after 24, 48 and 72 h of culture (24 h: t=2.667, 2.664; 48 h: t=4.559, 4.524; 72 h: t=7.257, 6.981; P<0.05). Compared with 24-h culture, the A value of MTT test in 48 and 72 hours in three groups were increased (48 h: t=5.507, 5.092, 3.741; 72 h: t=11.330, 10.637, 9.229; P<0.05), and the A value of MTT test in 72 hours were higher than those in 48 hours(t=7.411, 6.941, 5.214, P<0.05). The cell migration rate of Si-miR-182 group [(53.90±3.19)%] was lower than that of the control group [(81.66±5.92)%] and N-miR-182 group [(80.35±5.40)%] (t=9.231, 9.430, P<0.05). The relative expressions of SATB2, E-cadherin mRNA and protein in Si-miR-182 group were higher than those in control group and N-miR-182 group (SATB2: t=10.930, 11.158; E-Cadherin: t=9.288, 9.369; P<0.05), while the relative expressions of nox4, vimentin mRNA and protein in Si-miR-182 group were lower than those in control group and N-miR-182 group (nox4: t=8.955, 7.590; Vimentin: t=6.543, 6.644; P<0.05). Conclusion Silencing of miR-182 gene can significantly inhibit the proliferation and migration of colorectal cancer cells, possibly by activating SATB2, E-Cadherin expression, inhibiting the expression of nox4, Vimentin, and participating in the SATB2/nox4 pathway. Key words: MicroRNA-182; Special AT-rich sequence-binding protein 2; Colorectal cancer; Proliferation; Migration
{"title":"MicroRNA-182 regulates colon cancer cell proliferation and migration by targeting special AT-rich sequence-binding protein 2 gene","authors":"Xu Jizhong, Gui-xian Wang, W. Yuan, Quanbo Zhou, Shengyun Hu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.018","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.018","url":null,"abstract":"Objective \u0000To observe the effect of microRNA (miRNA, miR)-182 on the proliferation and migration of colorectal cancer cells by targeting special AT-rich sequence-binding protein 2 (SATB2) gene, and to explore its effect on SATB2/icotinamide adenine dinucleotide phosphate oxidase 4 (nox4) pathway. \u0000 \u0000 \u0000Methods \u0000HT-29 cells in logarithmic phase were transfected with Si-miR-182 and Control-si resectively, by liposome transfection, serving as Si-miR-182 group and N-miR-182 group, respectively. The expression of miR-182 gene, the cell proliferation and migration, the mRNA and protein expression of SATB2, nox4, E-cadherin and vimentin were examined. \u0000 \u0000 \u0000Results \u0000The relative expression of miR-182 gene in Si-miR-182 group (0.35±0.05) was lower than that in control group (1.24±0.26) and N-miR-182 group (1.20±0.25) (t=7.517, 7.455, P<0.05). The A values of methyl thiazol tetrazolium (MTT) test in Si-miR-182 group were lower than those in control group and N-miR-182 group after 24, 48 and 72 h of culture (24 h: t=2.667, 2.664; 48 h: t=4.559, 4.524; 72 h: t=7.257, 6.981; P<0.05). Compared with 24-h culture, the A value of MTT test in 48 and 72 hours in three groups were increased (48 h: t=5.507, 5.092, 3.741; 72 h: t=11.330, 10.637, 9.229; P<0.05), and the A value of MTT test in 72 hours were higher than those in 48 hours(t=7.411, 6.941, 5.214, P<0.05). The cell migration rate of Si-miR-182 group [(53.90±3.19)%] was lower than that of the control group [(81.66±5.92)%] and N-miR-182 group [(80.35±5.40)%] (t=9.231, 9.430, P<0.05). The relative expressions of SATB2, E-cadherin mRNA and protein in Si-miR-182 group were higher than those in control group and N-miR-182 group (SATB2: t=10.930, 11.158; E-Cadherin: t=9.288, 9.369; P<0.05), while the relative expressions of nox4, vimentin mRNA and protein in Si-miR-182 group were lower than those in control group and N-miR-182 group (nox4: t=8.955, 7.590; Vimentin: t=6.543, 6.644; P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Silencing of miR-182 gene can significantly inhibit the proliferation and migration of colorectal cancer cells, possibly by activating SATB2, E-Cadherin expression, inhibiting the expression of nox4, Vimentin, and participating in the SATB2/nox4 pathway. \u0000 \u0000 \u0000Key words: \u0000MicroRNA-182; Special AT-rich sequence-binding protein 2; Colorectal cancer; Proliferation; Migration","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"63-66"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44319666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.039
H. Yuan, Chunjiao Yu, Yujuan Zhang, Ziming Huang, Lihua Wu, Ailing Zhang, Xuezhong Gao, Zhiyong Luo
Objective To investigate the significance of HOX transcription antisense RNA (HOTAIR) expression in triple-negative breast cancer (TNBC) and explore the molecular modulation of HOTAIR on the progression of TNBC. Methods Sixty-four human TNBC tissues and their corresponding non-tumor tissues were collected, and the HOTAIR levels were detected by real-time quantitative polymerase chain reaction (Real-time PCR). The different HOTAIR levels in tumor and non-tumor tissues were compared. And then we analyzed its significance in TNBC with different pathological stages. In MDA-MB231 cells, the effects of down-regulation of HOTAIR by small interfering RNA (siRNA) on the proliferation (MTT) and apoptosis (Western blotting) were detected. The effects of down-regulation of HOTAIR on the expression of enhancer of zeste homolog2 (EZH2), the enzymatic subunit of ploycomb repressor complex (PRC2), were detected. SPSS19.0 statistical software was used for analysis. Results Compared with the non-tumor tissues, the HOTAIR expression was significantly increased in TNBC tumors (0.71±0.10 vs. 0.50±0.06, t=20.286, P<0.01), especially in TNBC with advanced stage (0.78±0.05 vs. 0.64±0.07, t=8.625, P<0.01). In TNBC cells, treatment of siEZH2 showed similar effects on the cell proliferation and apoptosis. In TNBC cells, EZH2 expression was down-regulated by siHOTAIR. However, HOTAIR expression had no significant change after siEZH2 treatment. Conclusion HOTAIR is highly expressed in TNBC and associated with poor prognosis. Our results suggest HOTAIR may induce the carcinogenesis in TNBC probably through regulating EZH2. Key words: Triple negative breast cancer; HOX transcription antisense RNA; Enhancer of zeste homolog 2
{"title":"Effects of HOX transcription antisense RNA on carcinogenesis and development of triple-negative breast cancer","authors":"H. Yuan, Chunjiao Yu, Yujuan Zhang, Ziming Huang, Lihua Wu, Ailing Zhang, Xuezhong Gao, Zhiyong Luo","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.039","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.039","url":null,"abstract":"Objective \u0000To investigate the significance of HOX transcription antisense RNA (HOTAIR) expression in triple-negative breast cancer (TNBC) and explore the molecular modulation of HOTAIR on the progression of TNBC. \u0000 \u0000 \u0000Methods \u0000Sixty-four human TNBC tissues and their corresponding non-tumor tissues were collected, and the HOTAIR levels were detected by real-time quantitative polymerase chain reaction (Real-time PCR). The different HOTAIR levels in tumor and non-tumor tissues were compared. And then we analyzed its significance in TNBC with different pathological stages. In MDA-MB231 cells, the effects of down-regulation of HOTAIR by small interfering RNA (siRNA) on the proliferation (MTT) and apoptosis (Western blotting) were detected. The effects of down-regulation of HOTAIR on the expression of enhancer of zeste homolog2 (EZH2), the enzymatic subunit of ploycomb repressor complex (PRC2), were detected. SPSS19.0 statistical software was used for analysis. \u0000 \u0000 \u0000Results \u0000Compared with the non-tumor tissues, the HOTAIR expression was significantly increased in TNBC tumors (0.71±0.10 vs. 0.50±0.06, t=20.286, P<0.01), especially in TNBC with advanced stage (0.78±0.05 vs. 0.64±0.07, t=8.625, P<0.01). In TNBC cells, treatment of siEZH2 showed similar effects on the cell proliferation and apoptosis. In TNBC cells, EZH2 expression was down-regulated by siHOTAIR. However, HOTAIR expression had no significant change after siEZH2 treatment. \u0000 \u0000 \u0000Conclusion \u0000HOTAIR is highly expressed in TNBC and associated with poor prognosis. Our results suggest HOTAIR may induce the carcinogenesis in TNBC probably through regulating EZH2. \u0000 \u0000 \u0000Key words: \u0000Triple negative breast cancer; HOX transcription antisense RNA; Enhancer of zeste homolog 2","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"134-136"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48124041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.055
Tian Xiangyong, Du Wenjing, W. Xiaoqiang, Chan Zhang, Zhi-wei Wang, G. Cao, T. Yan
{"title":"Role of ciprofloxacin in preemptive therapy of BK viruria in renal transplant recipients","authors":"Tian Xiangyong, Du Wenjing, W. Xiaoqiang, Chan Zhang, Zhi-wei Wang, G. Cao, T. Yan","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.055","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.055","url":null,"abstract":"","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"181-181"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42322714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.009
Qianying He, Jia Yu, Fangchao Mei, Yu Zhou, Xiaojiao Yang, Chen-yang Wang, Man Li, Weixing Wang
Objective To investigate whether obesity aggravates acute lung injury and its mechanism in acute necrotizing pancreatitis. Methods Twenty-four male Sprague-Dawley rats were randomly divided into a normal diet control group (N-control), a normal diet-ANP group (N-ANP), and a high-fat diet control group (H-control), and a high fat diet-ANP group (H-ANP). Serum amylase (AMY), lipase (LIP), triglyceride (TG) and total cholesterol (TC) were determined by automatic biochemical analyzer. CD68, CD11c, CD206, toll like receptor-4 (TLR4), myeloperoxidase (MPO), interleukin (IL)-1β and nuclear factor-κB (NF-κB) in the lung tissue were detected by immunofluorescence or immunohistochemistry. The pancreas and lung tissues were observed under microscope and scored. SPSS 22.0 software was used. The data were expressed as mean±standard deviation. The t test, one-way ANOVA and SNK test were used. Results The TG and TC of the H-control group were (1.00±0.36) and (2.51±0.41) U/L, which were higher than those of the N-control group (0.53±0.19) and (1.72±0.52) U/L. The differences were statistically significant (t=-3.288, -3.062, P<0.05); AMY and LIP of the N-ANP group were (8 690.00±1 951.35), (9 650.00±1 810.19) U/L, which were higher than the N-control group (2 018.50±382.05), (90.26±7.00) U/L, the difference was statistically significant (q=9.452, 18.090, P<0.05); the LIP of the H-ANP group was (39 705.75±1 345.55) U/L, which was higher than the N-ANP group (q=59.940, P<0.05). The pancreas and lung scores in the N-ANP group were (11.40±1.80) and (6.62±1.06) points, which were higher than those in the N-control group (0.31±0.29) and (0.75±0.88). The difference was statistically significant (q=21.950, 17.160, P<0.05). The lung score of the H-ANP group was (8.19±1.07) points, which was higher than that of the N-ANP group (q=4.017, P<0.05), and the difference was statistically significant. The lung NF-κB of N-ANP group was (0.38±0.02), which was higher than that of N-control group (0.23±0.02) and lower than that of H-ANP group (0.48±0.01). The difference was statistically significant (q=21.830, 13.030, P<0.05). The number of IL-1β, MPO, CD68+ , CD68+ /CD11C+ , CD68+ /TLR4+ cells in the N-ANP group was (43.80±4.29), (105.81±5.37), (34.17±3.27), (13.32±1.83), (18.43±1.06), higher than the N-control group (4.93±1.13), (10.37±1.77), (20.16±2.42), (1.54±0.75), (8.37±1.40), the difference was statistically significant (q=29.530, 51.010, 15.261, 17.952, 20.965, P<0.05), lower than the H-ANP group (92.58±5.83), (175.71±8.85), (41.61±2.56), (16.92±1.96), (22.91±2.10), the difference was statistically significant (q=36.820, 37.100, 8.176, 8.079, 6.467, P<0.05). The number of lung CD68+ /CD206+ cells in the N-ANP group was (20.43±1.30), which was higher than that in the N-control group (11.65±1.51) and H-ANP group (14.83±1.83). The difference was statistically significant (q=16.613, 10.682, P<0.05). Conclusion Obesity can aggravate ALI in ANP, which may be rel
{"title":"Role of macrophage polarization in lung injury in obese rats with acute necrotizing pancreatitis","authors":"Qianying He, Jia Yu, Fangchao Mei, Yu Zhou, Xiaojiao Yang, Chen-yang Wang, Man Li, Weixing Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.009","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.009","url":null,"abstract":"Objective \u0000To investigate whether obesity aggravates acute lung injury and its mechanism in acute necrotizing pancreatitis. \u0000 \u0000 \u0000Methods \u0000Twenty-four male Sprague-Dawley rats were randomly divided into a normal diet control group (N-control), a normal diet-ANP group (N-ANP), and a high-fat diet control group (H-control), and a high fat diet-ANP group (H-ANP). Serum amylase (AMY), lipase (LIP), triglyceride (TG) and total cholesterol (TC) were determined by automatic biochemical analyzer. CD68, CD11c, CD206, toll like receptor-4 (TLR4), myeloperoxidase (MPO), interleukin (IL)-1β and nuclear factor-κB (NF-κB) in the lung tissue were detected by immunofluorescence or immunohistochemistry. The pancreas and lung tissues were observed under microscope and scored. SPSS 22.0 software was used. The data were expressed as mean±standard deviation. The t test, one-way ANOVA and SNK test were used. \u0000 \u0000 \u0000Results \u0000The TG and TC of the H-control group were (1.00±0.36) and (2.51±0.41) U/L, which were higher than those of the N-control group (0.53±0.19) and (1.72±0.52) U/L. The differences were statistically significant (t=-3.288, -3.062, P<0.05); AMY and LIP of the N-ANP group were (8 690.00±1 951.35), (9 650.00±1 810.19) U/L, which were higher than the N-control group (2 018.50±382.05), (90.26±7.00) U/L, the difference was statistically significant (q=9.452, 18.090, P<0.05); the LIP of the H-ANP group was (39 705.75±1 345.55) U/L, which was higher than the N-ANP group (q=59.940, P<0.05). The pancreas and lung scores in the N-ANP group were (11.40±1.80) and (6.62±1.06) points, which were higher than those in the N-control group (0.31±0.29) and (0.75±0.88). The difference was statistically significant (q=21.950, 17.160, P<0.05). The lung score of the H-ANP group was (8.19±1.07) points, which was higher than that of the N-ANP group (q=4.017, P<0.05), and the difference was statistically significant. The lung NF-κB of N-ANP group was (0.38±0.02), which was higher than that of N-control group (0.23±0.02) and lower than that of H-ANP group (0.48±0.01). The difference was statistically significant (q=21.830, 13.030, P<0.05). The number of IL-1β, MPO, CD68+ , CD68+ /CD11C+ , CD68+ /TLR4+ cells in the N-ANP group was (43.80±4.29), (105.81±5.37), (34.17±3.27), (13.32±1.83), (18.43±1.06), higher than the N-control group (4.93±1.13), (10.37±1.77), (20.16±2.42), (1.54±0.75), (8.37±1.40), the difference was statistically significant (q=29.530, 51.010, 15.261, 17.952, 20.965, P<0.05), lower than the H-ANP group (92.58±5.83), (175.71±8.85), (41.61±2.56), (16.92±1.96), (22.91±2.10), the difference was statistically significant (q=36.820, 37.100, 8.176, 8.079, 6.467, P<0.05). The number of lung CD68+ /CD206+ cells in the N-ANP group was (20.43±1.30), which was higher than that in the N-control group (11.65±1.51) and H-ANP group (14.83±1.83). The difference was statistically significant (q=16.613, 10.682, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Obesity can aggravate ALI in ANP, which may be rel","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"29-32"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47188717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.025
Bowen Liu, Peng Li, Chaohui Gu, Zhankui Jia, Jinjian Yang
Objective To establish a mouse model of neurogenic bladder and to investigate the changes of fibronectin 1 (FN1) and the action mechanism of microRNA (miRNA, miR)-1a-3p. Methods Mice were divided into model group and control group. The model group received subcutaneous injection of myelin oligodendrocyte glycoprotein and mycobacterium tuberculosis, and intraperitoneal injection of pertussis toxin at 48 h. The control group received subcutaneous injection of normal saline. The frequency of urination and the amount of urine output were observed. The mice were sacrificed and the bladder tissue was taken. Three bladders were selected from the model group and the control group for high-throughput sequencing. The changes of FN1 and miR-1a-3p in mouse bladder tissues were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. Results The sequencing results were analyzed. As the urinary system symptoms worsened, FN1 increased (4.569±0.426, t=-5.142, P<0.01), while miR-1a-3p decreased (1.623±0.312, t=-3.945, P<0.05). Compared with the control group, the expression of FN1 mRNA [2.501 (1.301, 4.251), F=99.183, P<0.01] and protein [2.937 (1.174, 4.262), F=63.834, P<0.01] in the bladder tissue of the model group was up-regulated, and that of miR-1a-3p was down-regulated [2.401 (1.250, 3.001), F=35.951, P<0.01]. The dual luciferase gene report indicated that FN1 was a direct target gene of miR-1a-3p (0.514±0.027, t=13.355, P<0.01). Conclusion The animal model of neurogenic bladder has urinary frequency, urgency, urinary retention and other symptoms, and the expression of FN1 is up-regulated, while the expression of miR-1a-3p is down-regulated. Therefore, miR-1a-3p may participate in neurogenic bladder formation through regulation of FN1. Key words: Neurogenic bladder; Fibronectin 1; MicroRNA-1a-3p
{"title":"Action mechanism of microRNA-1a-3p in mouse neurogenic bladder","authors":"Bowen Liu, Peng Li, Chaohui Gu, Zhankui Jia, Jinjian Yang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.025","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.025","url":null,"abstract":"Objective \u0000To establish a mouse model of neurogenic bladder and to investigate the changes of fibronectin 1 (FN1) and the action mechanism of microRNA (miRNA, miR)-1a-3p. \u0000 \u0000 \u0000Methods \u0000Mice were divided into model group and control group. The model group received subcutaneous injection of myelin oligodendrocyte glycoprotein and mycobacterium tuberculosis, and intraperitoneal injection of pertussis toxin at 48 h. The control group received subcutaneous injection of normal saline. The frequency of urination and the amount of urine output were observed. The mice were sacrificed and the bladder tissue was taken. Three bladders were selected from the model group and the control group for high-throughput sequencing. The changes of FN1 and miR-1a-3p in mouse bladder tissues were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. \u0000 \u0000 \u0000Results \u0000The sequencing results were analyzed. As the urinary system symptoms worsened, FN1 increased (4.569±0.426, t=-5.142, P<0.01), while miR-1a-3p decreased (1.623±0.312, t=-3.945, P<0.05). Compared with the control group, the expression of FN1 mRNA [2.501 (1.301, 4.251), F=99.183, P<0.01] and protein [2.937 (1.174, 4.262), F=63.834, P<0.01] in the bladder tissue of the model group was up-regulated, and that of miR-1a-3p was down-regulated [2.401 (1.250, 3.001), F=35.951, P<0.01]. The dual luciferase gene report indicated that FN1 was a direct target gene of miR-1a-3p (0.514±0.027, t=13.355, P<0.01). \u0000 \u0000 \u0000Conclusion \u0000The animal model of neurogenic bladder has urinary frequency, urgency, urinary retention and other symptoms, and the expression of FN1 is up-regulated, while the expression of miR-1a-3p is down-regulated. Therefore, miR-1a-3p may participate in neurogenic bladder formation through regulation of FN1. \u0000 \u0000 \u0000Key words: \u0000Neurogenic bladder; Fibronectin 1; MicroRNA-1a-3p","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"87-89"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48629315","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.024
Wei-Tein Chang, R. Sun, Min Feng, Yuexia Li, Cui Zhiwen, Yi-lei Deng
Objective To investigate the effect of mitogen-activated protein kinase phosphatase 1 (MKP1) on tumor necrosis factor-α (TNF-α)-induced myocardial injury. Methods Wild type (WT) and MKP1 transgene (MKP1) mice were randomly divided into 2 groups separately: WT group, WT+ TNF-α group and MKP1 group, MKP1+ TNF-α group. Mice in the WT+ TNF-α group and the MKP1+ TNF-α group were intraperitoneally injected with TNF-α at a dose of 6 mg/kg. Mice in the WT group and the MKP1 group were intraperitoneally injected with an equal amount of physiological saline. The MKP1 expression, myocardial injury marker level, myocardial mitochondrial division-associated protein 1 (Drp1), antioxidant and respiratory complex expression and mitochondrial apoptosis were measured, and t-test was used to analyze differences between groups. Results As compared with WT+ TNF-α group, MKP1+ TNF-α group showed down-regulation of Drp1 and Mff expression (Drp1: 1.80±0.20 vs. 1.00±0.30, t=-10.134, P<0.05; Mff: 2.80±0.20 vs. 1.10±0.30, t=-8.313, P<0.05), increased expression of glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPX) [GSH: (29±2) vs. (49±3) nmol/mg, t=12.127, P<0.05; SOD: (2.2±0.1) vs. (8.1±0.2) U/mg, t=10.301, P<0.05; GPX: (50±4) vs. (172±6) U/mg, t=11.136, P<0.05], increased expression of mitochondrial respiratory recombination Ⅲ and Ⅱ (complex Ⅲ: 1.00±0.20 vs. 2.20±0.12, t=10.715, P<0.05; complex Ⅱ: 1.10±0.09 vs. 1.90±0.08, t=8.312, P<0.05), down-regulation of Caspase-9 and bax expression (Caspase-9: 2.20±0.11 vs. 1.15±0.09, t=-5.210, P<0.05; bax: 2.30±0.12 vs. 1.42±0.09, t=-6.006, P<0.05). Conclusion The mechanism of TNF-α-induced myocardial injury involves excessive division of mitochondria, mitochondrial redox balance, destruction of energy metabolism and mitochondrial apoptosis. MKP1 overexpression can significantly inhibit the development of these adverse reactions, protect mitochondrial function and inhibit myocardial damage. Key words: Mitogen-activated protein kinase phosphatase 1; Tumor necrosis factor-α; Myocardial injury
{"title":"Protective effect of mitogen-activated protein kinase phosphatase 1 on tumor necrosis factor-α-mediated myocardial injury","authors":"Wei-Tein Chang, R. Sun, Min Feng, Yuexia Li, Cui Zhiwen, Yi-lei Deng","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.024","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.024","url":null,"abstract":"Objective \u0000To investigate the effect of mitogen-activated protein kinase phosphatase 1 (MKP1) on tumor necrosis factor-α (TNF-α)-induced myocardial injury. \u0000 \u0000 \u0000Methods \u0000Wild type (WT) and MKP1 transgene (MKP1) mice were randomly divided into 2 groups separately: WT group, WT+ TNF-α group and MKP1 group, MKP1+ TNF-α group. Mice in the WT+ TNF-α group and the MKP1+ TNF-α group were intraperitoneally injected with TNF-α at a dose of 6 mg/kg. Mice in the WT group and the MKP1 group were intraperitoneally injected with an equal amount of physiological saline. The MKP1 expression, myocardial injury marker level, myocardial mitochondrial division-associated protein 1 (Drp1), antioxidant and respiratory complex expression and mitochondrial apoptosis were measured, and t-test was used to analyze differences between groups. \u0000 \u0000 \u0000Results \u0000As compared with WT+ TNF-α group, MKP1+ TNF-α group showed down-regulation of Drp1 and Mff expression (Drp1: 1.80±0.20 vs. 1.00±0.30, t=-10.134, P<0.05; Mff: 2.80±0.20 vs. 1.10±0.30, t=-8.313, P<0.05), increased expression of glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPX) [GSH: (29±2) vs. (49±3) nmol/mg, t=12.127, P<0.05; SOD: (2.2±0.1) vs. (8.1±0.2) U/mg, t=10.301, P<0.05; GPX: (50±4) vs. (172±6) U/mg, t=11.136, P<0.05], increased expression of mitochondrial respiratory recombination Ⅲ and Ⅱ (complex Ⅲ: 1.00±0.20 vs. 2.20±0.12, t=10.715, P<0.05; complex Ⅱ: 1.10±0.09 vs. 1.90±0.08, t=8.312, P<0.05), down-regulation of Caspase-9 and bax expression (Caspase-9: 2.20±0.11 vs. 1.15±0.09, t=-5.210, P<0.05; bax: 2.30±0.12 vs. 1.42±0.09, t=-6.006, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The mechanism of TNF-α-induced myocardial injury involves excessive division of mitochondria, mitochondrial redox balance, destruction of energy metabolism and mitochondrial apoptosis. MKP1 overexpression can significantly inhibit the development of these adverse reactions, protect mitochondrial function and inhibit myocardial damage. \u0000 \u0000 \u0000Key words: \u0000Mitogen-activated protein kinase phosphatase 1; Tumor necrosis factor-α; Myocardial injury","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"84-86"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44794479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To isolate extracellular vesicles (VEs) from the culture supernatant of pancreatic cancer (PANC-1) and human pancreatic ductal epithelial cells (HPDE6-C7), and to compare the differential expression of microRNA (miRNA, miR)-483-5p in the two kinds of cells and their secreted EVs. Methods Ultracentrifugation method was used to prepare the complete culture medium of depletion of serum-derived vesicles. Cell count Kit (CCK-8) was used to detect the effect of complete medium on cell proliferation without ultracentrifugation (control group) and with ultracentrifugation (experimental group). Cells were cultured in vesicle-depleted complete medium, and EVs were extracted by ultracentrifugation. The concentration of EVs was detected by BCA protein concentration determination kit and nanoparticles tracking analysis (NTA). Transmission electron microscope, NTA, and Western blotting were used to identify whether they, obtained by ultracentrifugation, were in accordance with the characteristics of EVs. The expression of miR-483-5p was detected by quantitative real time polymerase chain reaction (qPCR). The results of CCK-8 proliferation test and qPCR test were expressed by mean±standard deviation (SD). The statistical method was t test. Results The results of CCK-8 proliferation test showed that there was no significant difference between the experimental group and the control group after 48 h and 72 h in HPDE6-C7 cells (HPDE6-C7, 0.674±0.036 vs. 0.671±0.016, t=0.315, P48 h>0.05; 0.890±0.027 vs. 0.925±0.099, t=0.581, P72 h>0.05), as well as the PANC-1 (0.759±0.004 vs. 0.761±0.016, t=0.249, P48 h>0.05; 1.114±0.025 vs. 1.145±0.014, t=1.898, P72 h>0.05). The transmission electron microscopy showed that the EVs were "tea-cup tray" . The results of NTA showed that the diameter of 98% PANC-1 derived EVs was 130.0 nm, and that of 98% HPDE6-C7 derived EVs was 129.7 nm. Western blotting showed that CD63 and TSG101 were positive, but GM130 was negative. The concentration of EVs secreted by PANC-1 and HPDE6-C7 was measured by protein concentration assay kit and NTA as follows: 1.5 g/L, 1.510 11 particles/mland 1.3 g/L, 1.610 11 particles/ml, respectively. Compared with HPDE6-C7, the expression of miR-483-5p in PANC-1 was significantly increased (2.820±0.180 vs. 1.000±0.006, t=-17.539, P<0.01). Compared with HPDE6-C7 derived EVs, the expression of miR-483-5p in PANC-1 derived EVs was significantly increased (3.503±0.265 vs. 1.002±0.084, t=-15.582, P<0.01). Conclusion The ultracentrifugation method does not affect the normal proliferation of cells while removing fetal bovine serum-derived vesicles in the complete medium; the identification results show that the isolated pellets correspond to the characteristics of EVs. According to the results of NTA, it can be judged that they should be classified as small EVs. The results of qPCR show that miR-483-5p is highly expressed in PANC-1 and the secreted EVs. Key words: Pancreatic cancer; Ext
{"title":"Basic research of pancreatic cancer cell-derived extracellular vesicles","authors":"Jinxiang Xu, Shanglong Liu, Yuqi Sun, Zequn Li, Hengjian Liu, Dan Zhang, Yuanzhong Ren, Yanbing Zhou","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.003","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.003","url":null,"abstract":"Objective \u0000To isolate extracellular vesicles (VEs) from the culture supernatant of pancreatic cancer (PANC-1) and human pancreatic ductal epithelial cells (HPDE6-C7), and to compare the differential expression of microRNA (miRNA, miR)-483-5p in the two kinds of cells and their secreted EVs. \u0000 \u0000 \u0000Methods \u0000Ultracentrifugation method was used to prepare the complete culture medium of depletion of serum-derived vesicles. Cell count Kit (CCK-8) was used to detect the effect of complete medium on cell proliferation without ultracentrifugation (control group) and with ultracentrifugation (experimental group). Cells were cultured in vesicle-depleted complete medium, and EVs were extracted by ultracentrifugation. The concentration of EVs was detected by BCA protein concentration determination kit and nanoparticles tracking analysis (NTA). Transmission electron microscope, NTA, and Western blotting were used to identify whether they, obtained by ultracentrifugation, were in accordance with the characteristics of EVs. The expression of miR-483-5p was detected by quantitative real time polymerase chain reaction (qPCR). The results of CCK-8 proliferation test and qPCR test were expressed by mean±standard deviation (SD). The statistical method was t test. \u0000 \u0000 \u0000Results \u0000The results of CCK-8 proliferation test showed that there was no significant difference between the experimental group and the control group after 48 h and 72 h in HPDE6-C7 cells (HPDE6-C7, 0.674±0.036 vs. 0.671±0.016, t=0.315, P48 h>0.05; 0.890±0.027 vs. 0.925±0.099, t=0.581, P72 h>0.05), as well as the PANC-1 (0.759±0.004 vs. 0.761±0.016, t=0.249, P48 h>0.05; 1.114±0.025 vs. 1.145±0.014, t=1.898, P72 h>0.05). The transmission electron microscopy showed that the EVs were \"tea-cup tray\" . The results of NTA showed that the diameter of 98% PANC-1 derived EVs was 130.0 nm, and that of 98% HPDE6-C7 derived EVs was 129.7 nm. Western blotting showed that CD63 and TSG101 were positive, but GM130 was negative. The concentration of EVs secreted by PANC-1 and HPDE6-C7 was measured by protein concentration assay kit and NTA as follows: 1.5 g/L, 1.510 11 particles/mland 1.3 g/L, 1.610 11 particles/ml, respectively. Compared with HPDE6-C7, the expression of miR-483-5p in PANC-1 was significantly increased (2.820±0.180 vs. 1.000±0.006, t=-17.539, P<0.01). Compared with HPDE6-C7 derived EVs, the expression of miR-483-5p in PANC-1 derived EVs was significantly increased (3.503±0.265 vs. 1.002±0.084, t=-15.582, P<0.01). \u0000 \u0000 \u0000Conclusion \u0000The ultracentrifugation method does not affect the normal proliferation of cells while removing fetal bovine serum-derived vesicles in the complete medium; the identification results show that the isolated pellets correspond to the characteristics of EVs. According to the results of NTA, it can be judged that they should be classified as small EVs. The results of qPCR show that miR-483-5p is highly expressed in PANC-1 and the secreted EVs. \u0000 \u0000 \u0000Key words: \u0000Pancreatic cancer; Ext","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"12-14"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42919169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.033
Xiang Yu, D. Ren, W. Feng, Qiong Han, Die Hu
Objective To investigate the mechanism of overexpression of breast cancer metastasis suppressor 1 (BRMS1) gene on the sensitivity of paclitaxel in osteosarcoma. Methods Human osteosarcoma MG-63 cells were divided into blank group, pcDNA3.1-BRMS1 group, paclitaxel group and pcDNA3.1-BRMS1 + paclitaxel group. The blank plasmid pcDNA3.1 (+ ) and recombinant plasmid pcDNA3.1 (+ )-BRMS1 were transfected into MG-63 cells by Lipofectamine™ 2000. Western blotting was used to detect the expression of BRMS1, nuclear factor-κB (NF-κB) p65, proliferating cell nuclear antigen (PCNA) and B cell lymphoma/leukemia-2 associated X protein (bax) protein, while methyl thiazol tetrazolium (MTT) assay and Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) double staining were used to detect cell viability and apoptosis rate, respectively. SPSS 21.0 statistical software was used for analysis. The measurement data were expressed as Mean± standard deviation (Mean±SD). T-test was used for inter-group comparison. Results The expression of BRMS1 protein in MG-63 cells after pcDNA3.1-BRMS1 transfection (0.216±0.018) was significantly higher than that in the blank group (0.018±0.004, t=33.956, P<0.05). Compared with the blank group, cell viability in pcDNA3.1-BRMS1 group and paclitaxel group (0.603±0.051, 0.536±0.04) decreased significantly (t=8.311, 11.914, P<0.05), apoptosis rate [(17.18±0.89)%, (20.02±1.07)%] increased significantly (t=48.437, 48.924, P<0.05), expression of NF-κB p65 (0.307±0.029, 0.257±0.026) decreased significantly (t=16.580, 19.403, P<0.05), expression of PCNA protein (0.222±0.021, 0.167±0.016) decreased significantly (t=7.121, 12.203, P<0.05), and expression of Bax protein (0.091±0.012, 0.131±0.013) increased significantly (t=14.352, 22.476, P<0.05). The combined use of pcDNA3.1-BRMS1 and paclitaxel had significant effects on cell viability, apoptosis and expression of NF-κB p65, PCNA and bax protein. Conclusion Overexpression of BRMS1 can inhibit the viability of osteosarcoma cells, induce apoptosis and enhance the sensitivity of paclitaxel. The mechanism is related to inhibition of NF-κB signaling pathway. Key words: Osteosarcoma; Breast cancer metastasis suppressor 1 gene; Paclitaxel sensitivity; Nuclear factor-κB signaling pathway
{"title":"Breast cancer metastasis suppressor 1 gene enhances the sensitivity of osteosarcoma cells to paclitaxel","authors":"Xiang Yu, D. Ren, W. Feng, Qiong Han, Die Hu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.033","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.033","url":null,"abstract":"Objective \u0000To investigate the mechanism of overexpression of breast cancer metastasis suppressor 1 (BRMS1) gene on the sensitivity of paclitaxel in osteosarcoma. \u0000 \u0000 \u0000Methods \u0000Human osteosarcoma MG-63 cells were divided into blank group, pcDNA3.1-BRMS1 group, paclitaxel group and pcDNA3.1-BRMS1 + paclitaxel group. The blank plasmid pcDNA3.1 (+ ) and recombinant plasmid pcDNA3.1 (+ )-BRMS1 were transfected into MG-63 cells by Lipofectamine™ 2000. Western blotting was used to detect the expression of BRMS1, nuclear factor-κB (NF-κB) p65, proliferating cell nuclear antigen (PCNA) and B cell lymphoma/leukemia-2 associated X protein (bax) protein, while methyl thiazol tetrazolium (MTT) assay and Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) double staining were used to detect cell viability and apoptosis rate, respectively. SPSS 21.0 statistical software was used for analysis. The measurement data were expressed as Mean± standard deviation (Mean±SD). T-test was used for inter-group comparison. \u0000 \u0000 \u0000Results \u0000The expression of BRMS1 protein in MG-63 cells after pcDNA3.1-BRMS1 transfection (0.216±0.018) was significantly higher than that in the blank group (0.018±0.004, t=33.956, P<0.05). Compared with the blank group, cell viability in pcDNA3.1-BRMS1 group and paclitaxel group (0.603±0.051, 0.536±0.04) decreased significantly (t=8.311, 11.914, P<0.05), apoptosis rate [(17.18±0.89)%, (20.02±1.07)%] increased significantly (t=48.437, 48.924, P<0.05), expression of NF-κB p65 (0.307±0.029, 0.257±0.026) decreased significantly (t=16.580, 19.403, P<0.05), expression of PCNA protein (0.222±0.021, 0.167±0.016) decreased significantly (t=7.121, 12.203, P<0.05), and expression of Bax protein (0.091±0.012, 0.131±0.013) increased significantly (t=14.352, 22.476, P<0.05). The combined use of pcDNA3.1-BRMS1 and paclitaxel had significant effects on cell viability, apoptosis and expression of NF-κB p65, PCNA and bax protein. \u0000 \u0000 \u0000Conclusion \u0000Overexpression of BRMS1 can inhibit the viability of osteosarcoma cells, induce apoptosis and enhance the sensitivity of paclitaxel. The mechanism is related to inhibition of NF-κB signaling pathway. \u0000 \u0000 \u0000Key words: \u0000Osteosarcoma; Breast cancer metastasis suppressor 1 gene; Paclitaxel sensitivity; Nuclear factor-κB signaling pathway","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"115-117"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49510997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.007
Fan Jiang, Hongsong Xing, Yue Ruan, Wei-Y Chen, Guojun Wu, Anyi Lin, Hai Hu
Objective To investigate the role and mechanism of the activation of B cell lymphoma/leukemia-2-antagonist/killer 1 (bak1) signaling pathway in anticancer effects of itraconazole on pancreatic cancers. Methods The experiment was divided into control group and itraconazole group according to the different detection indicators. Human pancreatic cancer MIA PaCa-2 and CFPAC-1 cells were treated with different concentrations of itraconazole (0, 10, 30, 50, 70, 90 mg/L) bought from Merk’s company in USA. Cell proliferation was evaluated by 3-(4, 5-dimethylhiazol-2-yl)-3, 5-diphenyltetrazolium bromide (MTT) assay. Cell cycle was analyzed by flow cytometry, and cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL) immunofluorescence assay. Moreover, bak1 small interfering RNA (siRNA) was used to inhibit the expression of bak1, and then the proliferation and apoptosis of pancreatic cancer cells treated with itraconazole were evaluated. The expression of bak1 and its downstream bax and cysteinyl aspartate-specific protease (Caspase)-3 proteins was measured by Western blotting. SPSS 19.0 statistical software was used to analyze the measurement data. The mean soil standard deviation (Mean±SD) was used to express the measurement data. The analysis of variance was used for the comparison between groups. Results The inhibitory rates of 10, 30, 50, 70 and 90 mg/L itraconazole Mia PaCa-2 were 6.211%, 34.741%, 63.226%, 82.531% and 89.112% respectively, and the inhibitory rates of itraconazole on CFPAC-1 were 9.726%, 47.322%, 53.631%, 72.629% and 92.641% respectively. Before and after 50 μmol/L itraconazole intervention, Mia PaCa-2 G0/G1 phase was 43.142% and 57.341% (t=21.762, P<0.05), the difference was statistically significant; CFPAC-1 G0/G1 phase was 40.107% and 63.216% (t=17.186, P<0.05), the difference was statistically significant. 50 mg /L itraconazole could effectively induce the expression of bak1, Bax and cleaved caspase-3 in Mia PaCa-2 cells, the difference was statistically significant (3.406%, 10.712% and 22.626%, t=40.835, P<0.05), the difference was statistically significant (5.041%, 16.135% and 23.701%, t=36.761, P<0.05). After the inhibition of bak1 expression by bak1 siRNA, the inhibition of proliferation and apoptosis induced by itraconazole decreased significantly. Conclusion Itraconazole exerts anti pancreatic cancer effect by regulating the activation of bak1 signaling pathway. Key words: Itraconazole; Pancreatic cancer; Proliferation; Apoptosis; B cell lymphoma/leukemia-2-antagonist/killer 1
{"title":"Effects of itraconazole on proliferation and apoptosis of pancreatic cancer cells","authors":"Fan Jiang, Hongsong Xing, Yue Ruan, Wei-Y Chen, Guojun Wu, Anyi Lin, Hai Hu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.007","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.007","url":null,"abstract":"Objective \u0000To investigate the role and mechanism of the activation of B cell lymphoma/leukemia-2-antagonist/killer 1 (bak1) signaling pathway in anticancer effects of itraconazole on pancreatic cancers. \u0000 \u0000 \u0000Methods \u0000The experiment was divided into control group and itraconazole group according to the different detection indicators. Human pancreatic cancer MIA PaCa-2 and CFPAC-1 cells were treated with different concentrations of itraconazole (0, 10, 30, 50, 70, 90 mg/L) bought from Merk’s company in USA. Cell proliferation was evaluated by 3-(4, 5-dimethylhiazol-2-yl)-3, 5-diphenyltetrazolium bromide (MTT) assay. Cell cycle was analyzed by flow cytometry, and cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL) immunofluorescence assay. Moreover, bak1 small interfering RNA (siRNA) was used to inhibit the expression of bak1, and then the proliferation and apoptosis of pancreatic cancer cells treated with itraconazole were evaluated. The expression of bak1 and its downstream bax and cysteinyl aspartate-specific protease (Caspase)-3 proteins was measured by Western blotting. SPSS 19.0 statistical software was used to analyze the measurement data. The mean soil standard deviation (Mean±SD) was used to express the measurement data. The analysis of variance was used for the comparison between groups. \u0000 \u0000 \u0000Results \u0000The inhibitory rates of 10, 30, 50, 70 and 90 mg/L itraconazole Mia PaCa-2 were 6.211%, 34.741%, 63.226%, 82.531% and 89.112% respectively, and the inhibitory rates of itraconazole on CFPAC-1 were 9.726%, 47.322%, 53.631%, 72.629% and 92.641% respectively. Before and after 50 μmol/L itraconazole intervention, Mia PaCa-2 G0/G1 phase was 43.142% and 57.341% (t=21.762, P<0.05), the difference was statistically significant; CFPAC-1 G0/G1 phase was 40.107% and 63.216% (t=17.186, P<0.05), the difference was statistically significant. 50 mg /L itraconazole could effectively induce the expression of bak1, Bax and cleaved caspase-3 in Mia PaCa-2 cells, the difference was statistically significant (3.406%, 10.712% and 22.626%, t=40.835, P<0.05), the difference was statistically significant (5.041%, 16.135% and 23.701%, t=36.761, P<0.05). After the inhibition of bak1 expression by bak1 siRNA, the inhibition of proliferation and apoptosis induced by itraconazole decreased significantly. \u0000 \u0000 \u0000Conclusion \u0000Itraconazole exerts anti pancreatic cancer effect by regulating the activation of bak1 signaling pathway. \u0000 \u0000 \u0000Key words: \u0000Itraconazole; Pancreatic cancer; Proliferation; Apoptosis; B cell lymphoma/leukemia-2-antagonist/killer 1","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"25-28"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48577674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.043
Wenqu Jiang, Raorao Yuan, Bin Wu, K. Xiong, Le Biao, Huang Xiangqun, B. Wang, Y. Zhuo, Yan Zhang
Objective To study the expression characteristics of polycomb repressive complex 1 (PRC1) gene in glioma and its influence on the survival of patients, as well as the influence on glioma cells after regulating its expression in vitro, and to evaluate the clinical significance of PRC1 in glioma. Methods Through TCGA database glioma database mining PRC1 mRNA level in glioblastoma tumor tissue, the level of the differences between different level of the WHO, the types of cases, 56 cases of glioma tumor samples of immunohistochemical staining to evaluate PRC1 expression characteristics and the relationship between Ki67, 6 glioma cell line by comparing protein imprinting experiments with astrocytes in PRC1 protein expression level difference, Transcriptome data of patients with high expression of PRC1 and patients with low expression of PRC1 in TCGA database were analyzed to study the molecular signaling pathways that may be regulated by PRC1, and to explore the clinical significance of PRC1 in glioma. Results The higher the WHO level was, the higher PRC1 mRNA (PRC1 mRNA level: Grade Ⅲ vs. Grade Ⅱ: t=9.665, P<0.05; Grade Ⅳ vs. Grade Ⅲ: t=11.200, P<0.05; Grade Ⅳ vs. Grade Ⅱ: t=24.980, P<0.05; PRC1 IHC staining intensity level: Grade Ⅳ vs. Grade Ⅱ: t=8.120, P<0.05; Grade Ⅳ vs. Grade Ⅲ: t=5.957, P<0.05), and protein levels were, and the levels in glioblastoma were significantly higher than those in oligodendroglioma, oligodendroglioma, and astrocytoma (Oligodendroglioma vs. glioblastoma: t=16.110; oligodendroglioma vs. glioblastoma: t=15.460; astrocytoma vs. glioblastoma: t=13.290, all P<0.05; PRC1 IHC staining intensity level: F=20.540, P<0.05). Compared with normal brain tissue, PRC1 mRNA level in glioblastoma was significantly increased (t=6.432, P<0.05). The protein expression level of PRC1 in glioblastoma cell lines was significantly higher than that in normal astrocytes (U87MG vs. HA: t=3.797, U118MG vs. HA: t=5.008, U251 vs. HA: t=4.435, T98G vs. HA: t=5.867, A172 vs. HA: t=4.809, LN229 vs. HA: t=6.242, all P<0.05). The median survival time of patients in the PRC1 high-expression group was significantly lower than that in the PRC1 low-expression group, which was 13.3 months and 40.45 months, respectively (χ2=23.990, P<0.05). PRC1 was involved in the regulation of cell cycle and p53 signaling pathway, and there was a significantly positive correlation between PRC1 and proliferating cell nuclear antigen (Ki-67) (P<0.05). Conclusion PRC1 is abnormally highly expressed in glioma, and the tumor proliferation with high expression of PRC1 is significantly enhanced with higher malignant degree, indicating a poor clinical prognosis. PRC1 may be a new therapeutic target for glioma. Key words: Polycomb repressive complex 1; Glioma; Prognosis
目的研究多梳抑制复合体1 (polycomb suppressicomplex 1, PRC1)基因在胶质瘤中的表达特点及其对患者生存的影响,以及体外调节其表达后对胶质瘤细胞的影响,评价PRC1在胶质瘤中的临床意义。方法通过TCGA胶质瘤数据库挖掘胶质母细胞瘤肿瘤组织中PRC1 mRNA的表达水平,比较WHO不同水平、不同类型病例间的差异,对56例胶质瘤肿瘤样本进行免疫组化染色,评价PRC1表达特征及Ki67、6胶质瘤细胞系间的关系,通过蛋白印迹实验比较与星形胶质细胞中PRC1蛋白表达水平的差异。分析TCGA数据库中PRC1高表达患者和PRC1低表达患者的转录组数据,研究PRC1可能调控的分子信号通路,探讨PRC1在胶质瘤中的临床意义。结果WHO水平越高,PRC1 mRNA水平越高(PRC1 mRNA水平:分级Ⅲvs分级Ⅱ:t=9.665, P<0.05;分级Ⅳvs.Ⅲ:t=11.200, P<0.05;分级Ⅳvs.Ⅱ:t=24.980, P<0.05;PRC1 IHC染色强度水平:Ⅳ级vs.Ⅱ级:t=8.120, P<0.05;Ⅳ级vs.Ⅲ级:t=5.957, P<0.05),且胶质母细胞瘤的蛋白水平显著高于少突胶质细胞瘤、少突胶质细胞瘤和星形细胞瘤(少突胶质细胞瘤vs.胶质母细胞瘤:t=16.110;少突胶质细胞瘤vs.胶质母细胞瘤:t=15.460;星形细胞瘤vs胶质母细胞瘤:t=13.290, P均<0.05;PRC1 IHC染色强度水平:F=20.540, P<0.05)。与正常脑组织相比,胶质母细胞瘤组织中PRC1 mRNA水平显著升高(t=6.432, P<0.05)。PRC1蛋白在胶质母细胞瘤细胞系中的表达水平显著高于正常星形胶质细胞(U87MG vs. HA: t=3.797, U118MG vs. HA: t=5.008, U251 vs. HA: t=4.435, T98G vs. HA: t=5.867, A172 vs. HA: t=4.809, LN229 vs. HA: t=6.242,均P<0.05)。PRC1高表达组患者的中位生存时间显著低于PRC1低表达组,分别为13.3个月和40.45个月(χ2=23.990, P<0.05)。PRC1参与细胞周期和p53信号通路的调控,且与增殖细胞核抗原(Ki-67)呈显著正相关(P<0.05)。结论PRC1在胶质瘤中异常高表达,PRC1高表达的肿瘤增殖明显增强,恶性程度较高,临床预后较差。PRC1可能成为胶质瘤新的治疗靶点。关键词:Polycomb压抑复合体1;神经胶质瘤;预后
{"title":"Clinical implication and expression characteristics of polycomb repressive complex 1 in glioma","authors":"Wenqu Jiang, Raorao Yuan, Bin Wu, K. Xiong, Le Biao, Huang Xiangqun, B. Wang, Y. Zhuo, Yan Zhang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.043","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.043","url":null,"abstract":"Objective \u0000To study the expression characteristics of polycomb repressive complex 1 (PRC1) gene in glioma and its influence on the survival of patients, as well as the influence on glioma cells after regulating its expression in vitro, and to evaluate the clinical significance of PRC1 in glioma. \u0000 \u0000 \u0000Methods \u0000Through TCGA database glioma database mining PRC1 mRNA level in glioblastoma tumor tissue, the level of the differences between different level of the WHO, the types of cases, 56 cases of glioma tumor samples of immunohistochemical staining to evaluate PRC1 expression characteristics and the relationship between Ki67, 6 glioma cell line by comparing protein imprinting experiments with astrocytes in PRC1 protein expression level difference, Transcriptome data of patients with high expression of PRC1 and patients with low expression of PRC1 in TCGA database were analyzed to study the molecular signaling pathways that may be regulated by PRC1, and to explore the clinical significance of PRC1 in glioma. \u0000 \u0000 \u0000Results \u0000The higher the WHO level was, the higher PRC1 mRNA (PRC1 mRNA level: Grade Ⅲ vs. Grade Ⅱ: t=9.665, P<0.05; Grade Ⅳ vs. Grade Ⅲ: t=11.200, P<0.05; Grade Ⅳ vs. Grade Ⅱ: t=24.980, P<0.05; PRC1 IHC staining intensity level: Grade Ⅳ vs. Grade Ⅱ: t=8.120, P<0.05; Grade Ⅳ vs. Grade Ⅲ: t=5.957, P<0.05), and protein levels were, and the levels in glioblastoma were significantly higher than those in oligodendroglioma, oligodendroglioma, and astrocytoma (Oligodendroglioma vs. glioblastoma: t=16.110; oligodendroglioma vs. glioblastoma: t=15.460; astrocytoma vs. glioblastoma: t=13.290, all P<0.05; PRC1 IHC staining intensity level: F=20.540, P<0.05). Compared with normal brain tissue, PRC1 mRNA level in glioblastoma was significantly increased (t=6.432, P<0.05). The protein expression level of PRC1 in glioblastoma cell lines was significantly higher than that in normal astrocytes (U87MG vs. HA: t=3.797, U118MG vs. HA: t=5.008, U251 vs. HA: t=4.435, T98G vs. HA: t=5.867, A172 vs. HA: t=4.809, LN229 vs. HA: t=6.242, all P<0.05). The median survival time of patients in the PRC1 high-expression group was significantly lower than that in the PRC1 low-expression group, which was 13.3 months and 40.45 months, respectively (χ2=23.990, P<0.05). PRC1 was involved in the regulation of cell cycle and p53 signaling pathway, and there was a significantly positive correlation between PRC1 and proliferating cell nuclear antigen (Ki-67) (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000PRC1 is abnormally highly expressed in glioma, and the tumor proliferation with high expression of PRC1 is significantly enhanced with higher malignant degree, indicating a poor clinical prognosis. PRC1 may be a new therapeutic target for glioma. \u0000 \u0000 \u0000Key words: \u0000Polycomb repressive complex 1; Glioma; Prognosis","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"148-151"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44964501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}