Pub Date : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.025
Lu Liu, W. Lai, He-yang Xu, Yujie Zeng, Qiusheng Lan, Tao Zhang
Objective To investigate the effect of microRNA (miRNA, miR)-502-5p on colon cancer DLD-1 cells. Methods Colon cancer cell line DLD-1 was transfected by miR-502-5p mimic, control mimic, miR-502-5p inhibitor and control inhibitor respectively. The effect of miR-502-5p on the proliferation of DLD-1 cells was detected by cell counting kit-8 (CCK-8) assay. The effect of miR-502-5p on the colony forming ability of DLD-1 cells was detected by plate-cloning assay. Transwell assay was used to evaluate the effect of miR-502-5p on the migration and invasion of colon cells. Results The expression of miR-502-5p was significantly increased (319.31±33.16 vs. 1.08±0.22) in DLD-1 cells after transfection with miR-502-5p mimic (P 0.05), while migration [(621±90) cells vs. (174±32) cells] and invasion [(531±45) cells vs. (132±34) cells] ability were significantly increased (P 0.05), but the migration [(133±50) cells vs. (543±84) cells] and invasion [(119±45) cells vs. (458±57) cells] ability were decreased significantly (P<0.05). Conclusion MiR-502-5p may promote the migration and invasion of DLD-1 cells, but has no effect on the proliferation function. Key words: Colon cancer; MicroRNA-502-5p; Proliferation; Migration; Invasion
{"title":"The functional effects of microRNA-502-5p on colon cancer DLD-1 cells","authors":"Lu Liu, W. Lai, He-yang Xu, Yujie Zeng, Qiusheng Lan, Tao Zhang","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.025","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.025","url":null,"abstract":"Objective \u0000To investigate the effect of microRNA (miRNA, miR)-502-5p on colon cancer DLD-1 cells. \u0000 \u0000 \u0000Methods \u0000Colon cancer cell line DLD-1 was transfected by miR-502-5p mimic, control mimic, miR-502-5p inhibitor and control inhibitor respectively. The effect of miR-502-5p on the proliferation of DLD-1 cells was detected by cell counting kit-8 (CCK-8) assay. The effect of miR-502-5p on the colony forming ability of DLD-1 cells was detected by plate-cloning assay. Transwell assay was used to evaluate the effect of miR-502-5p on the migration and invasion of colon cells. \u0000 \u0000 \u0000Results \u0000The expression of miR-502-5p was significantly increased (319.31±33.16 vs. 1.08±0.22) in DLD-1 cells after transfection with miR-502-5p mimic (P 0.05), while migration [(621±90) cells vs. (174±32) cells] and invasion [(531±45) cells vs. (132±34) cells] ability were significantly increased (P 0.05), but the migration [(133±50) cells vs. (543±84) cells] and invasion [(119±45) cells vs. (458±57) cells] ability were decreased significantly (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000MiR-502-5p may promote the migration and invasion of DLD-1 cells, but has no effect on the proliferation function. \u0000 \u0000 \u0000Key words: \u0000Colon cancer; MicroRNA-502-5p; Proliferation; Migration; Invasion","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2207-2209"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49127244","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 explore the spatial-temporal expression of N-myc downstream-regulated gene 2 (NDRG2) in mouse brain after traumatic brain injury (TBI). Methods Male mice were randomly divided into the following groups: sham, TBI 6 h, TBI 12 h, TBI 24 h, TBI 48 h, TBI 72 h, TBI 7 d. A modified weight-drop method was utilized to induce mice TBI model. Western blot was used to detect the NDRG2 protein levels. Immunofluorescence was used to explore the cellular and subcellular locations of NDRG2. Student t test was used for difference comparison of NDRG2 protein levels between TBI and sham mice. Results The results of western blot showed that theNDRG2 protein levels increased significantly from 12 hours after TBI (1.52 fold, t=3.181, P<0.05), peaked at 24 hours (1.92 fold, t=5.032, P<0.01). The results of immunofluorescence showed that NDRG2 signals were specifically located in astrocytes. Besides, we observed NDRG2 nuclear translocation and co-localization with terminal deoxynucleotidyl transferase dUTPnick end labeling (TUNEL) signal safter TBI. Conclusion NDRG2 protein levels significantly increased after TBI. Besides, the increased proteins were partly translocated into nucleus and were co-localized with TUNEL signals, indicating that NDRG2 was involved with apoptosis after TBI. Key words: N-myc downstream-regulated gene 2; Traumatic brain injury; Spatial-temporal expression
{"title":"Spatial-temporal expression of N-myc downstream-regulated gene 2 in mouse brain after traumatic brain injury","authors":"Jiang Fang, Han-dong Wang, Yihao Zhu, Mao-xing Fei, Wenhao Niu, Xiao-liang Wang, Mengliang Zhou","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.031","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.031","url":null,"abstract":"Objective \u0000To explore the spatial-temporal expression of N-myc downstream-regulated gene 2 (NDRG2) in mouse brain after traumatic brain injury (TBI). \u0000 \u0000 \u0000Methods \u0000Male mice were randomly divided into the following groups: sham, TBI 6 h, TBI 12 h, TBI 24 h, TBI 48 h, TBI 72 h, TBI 7 d. A modified weight-drop method was utilized to induce mice TBI model. Western blot was used to detect the NDRG2 protein levels. Immunofluorescence was used to explore the cellular and subcellular locations of NDRG2. Student t test was used for difference comparison of NDRG2 protein levels between TBI and sham mice. \u0000 \u0000 \u0000Results \u0000The results of western blot showed that theNDRG2 protein levels increased significantly from 12 hours after TBI (1.52 fold, t=3.181, P<0.05), peaked at 24 hours (1.92 fold, t=5.032, P<0.01). The results of immunofluorescence showed that NDRG2 signals were specifically located in astrocytes. Besides, we observed NDRG2 nuclear translocation and co-localization with terminal deoxynucleotidyl transferase dUTPnick end labeling (TUNEL) signal safter TBI. \u0000 \u0000 \u0000Conclusion \u0000NDRG2 protein levels significantly increased after TBI. Besides, the increased proteins were partly translocated into nucleus and were co-localized with TUNEL signals, indicating that NDRG2 was involved with apoptosis after TBI. \u0000 \u0000 \u0000Key words: \u0000N-myc downstream-regulated gene 2; Traumatic brain injury; Spatial-temporal expression","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2227-2230"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46155954","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.038
Honggang Guo, S. Feng, Yuebin Zhou, Fang-Lian Yao, Yinzhong Liu, Zhi Chen, Y. Liang, Tao Wang
Objective To evaluate the interrelationship between intervertebral osteolysis and inflammatory or osteoclastic gene expression. Methods Adipose derived stromal cells (ADSCs) were osteoinduced, conventioanl tissue-engineered (TE) and surface-modified TE cage constructs were establisehd. Anterior cervical fusion model of goat was established. Implantation was carried out according to following six group: surface-modified TE cage group [Nd: YAG+ RGD+ β- tricalcium phosphate (β-TCP)/chitosan (CS)/polycaprolactone (PCL) cage+ ADSCs], TE cage group (β-TCP/CS/PCL cage+ ADSCs), non-modified β-TCP/CS/PCL cage, hydroxyapatite (HA) cage, titanium and bone allograft cage. Three-dimensional CT was used to observe morphology changing, bone mineralization ratio, new bone mension and osteoblatic quantity, interleukin-6 (IL-6), metalloprotease-1 (MMP-1), tumor necrosis factor-α (TNF-α) and tartrate resistant acid phosphatase (TRAP), and osteoprotein (OPG) gene expression were detected at 2, 4, 8, 12, 16, 20, 24, 28 and 36 weeks. Biomechanical property was also detected at different interval. Results Compared with other groups, implant subsidence and migration were not found in surface-modified TE cage group. Meanwhile, new bone prolifreated remarkably with satisfactory data of bone mineralization ratio, and new bone mension or osteoblatic quantity [(63.9±1.3)%, (76.54±1.8)%, (73.8±2.3) cells]. IL-6, MMP-1, TNF-α and TRAP gene downregulated (0.09±0.01, 0.13±0.02, 0.08±0.01). Conversely, Runx2 and OPG gene upregulated (0.73±0.13, 0.57±0.16). In addition, data of bone strength and rang of motion in surface-modified TE cage group were significantly higher than those of other groups [(4.38±0.25) Nm/degree, (6.55±0.19)°]. Conclusion Invertebral osteolysis can be effectively controlled by surface-modified TE approach, and this provides a potential therapy for implant aseptic loosening. Key words: Osteolysis; Cage; Surface modification; Tissue engineering; Interface
{"title":"The preliminary investigation on mechanism of preventing spinal interface osteolysis via surface-modified tissue-engineered cage","authors":"Honggang Guo, S. Feng, Yuebin Zhou, Fang-Lian Yao, Yinzhong Liu, Zhi Chen, Y. Liang, Tao Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.038","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.038","url":null,"abstract":"Objective \u0000To evaluate the interrelationship between intervertebral osteolysis and inflammatory or osteoclastic gene expression. \u0000 \u0000 \u0000Methods \u0000Adipose derived stromal cells (ADSCs) were osteoinduced, conventioanl tissue-engineered (TE) and surface-modified TE cage constructs were establisehd. Anterior cervical fusion model of goat was established. Implantation was carried out according to following six group: surface-modified TE cage group [Nd: YAG+ RGD+ β- tricalcium phosphate (β-TCP)/chitosan (CS)/polycaprolactone (PCL) cage+ ADSCs], TE cage group (β-TCP/CS/PCL cage+ ADSCs), non-modified β-TCP/CS/PCL cage, hydroxyapatite (HA) cage, titanium and bone allograft cage. Three-dimensional CT was used to observe morphology changing, bone mineralization ratio, new bone mension and osteoblatic quantity, interleukin-6 (IL-6), metalloprotease-1 (MMP-1), tumor necrosis factor-α (TNF-α) and tartrate resistant acid phosphatase (TRAP), and osteoprotein (OPG) gene expression were detected at 2, 4, 8, 12, 16, 20, 24, 28 and 36 weeks. Biomechanical property was also detected at different interval. \u0000 \u0000 \u0000Results \u0000Compared with other groups, implant subsidence and migration were not found in surface-modified TE cage group. Meanwhile, new bone prolifreated remarkably with satisfactory data of bone mineralization ratio, and new bone mension or osteoblatic quantity [(63.9±1.3)%, (76.54±1.8)%, (73.8±2.3) cells]. IL-6, MMP-1, TNF-α and TRAP gene downregulated (0.09±0.01, 0.13±0.02, 0.08±0.01). Conversely, Runx2 and OPG gene upregulated (0.73±0.13, 0.57±0.16). In addition, data of bone strength and rang of motion in surface-modified TE cage group were significantly higher than those of other groups [(4.38±0.25) Nm/degree, (6.55±0.19)°]. \u0000 \u0000 \u0000Conclusion \u0000Invertebral osteolysis can be effectively controlled by surface-modified TE approach, and this provides a potential therapy for implant aseptic loosening. \u0000 \u0000 \u0000Key words: \u0000Osteolysis; Cage; Surface modification; Tissue engineering; Interface","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2250-2253"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44265186","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.022
Jin-lu Liu, K. Zhao, Y. Qiu, Zhen Wang, Yuan-tian Mao, Bo-pei Li, Yeyang Chen
Objective To establish a cisplatin-resistant gastric cancer cell line MGC-803/cisplatin (DDP) and to explore its drug resistance mechanism. Methods The cisplatin was used to induce the drug resistance of MGC-803 cells. The half maximal inhibitory concentration (IC50) of the drug-resistant strains was detected by the cell counting kit-8 (CCK-8) method. The expression of chloride intracellular channel 1 (CLIC1) was detected by Western blotting analyses. The wild-type CLIC1 plasmid and the short hairpin RNA (shRNA) plasmid targeting CLIC1 were constructed. After Lipo3000 was transfected into gastric cancer cells, the IC50 of gastric cancer cells to cisplatin was detected, and then the intracellular Cl-concentration was detected by MQAE fluorescent probe. Results The cisplatin-resistant cell line MGC-803/DDP was successfully induced with an IC50 of (7.02±0.13) mg/L, while the IC50 of MGC803 was (1.29±0.09) mg/L, (t=36.090, P<0.05). The CLIC1 protein of MGC803/DDP was up-regulated by (2.27±0.15) times relative to MGC803 (t=7.841, P<0.05). After silencing the CLIC1 gene of MGC803/DDP, the IC50 of the CON, CN, and KD groups were (6.96±0.09), (6.93±0.15) and (3.02±0.20) mg/L, respectively, which was significantly decreased in the KD group (F=0.209, P<0.05). After overexpression of the CLIC1 gene in MGC803 cells, the IC50 of the CON, CN, and OE groups were (1.35±0.07), (1.25±0.07) and (4.77±0.12) mg/L, respectively, and the OE group was significantly elevated (F=0.508, P<0.05). The concentration of chloride ion was detected by MQAE. The relative fluorescence intensity of MGC-803, MGC-803/OE, MGC-803/DDP and MGC-803/DDP-KD were (1.02±0.03), (0.61±0.02), (0.67±0.01) and (1.39±0.02), respectively. The concentration of Cl- in MGC-803/OE cells was higher than that in MGC-803 (t=10.800, P<0.05), while the concentration of Cl- in MGC-803/DDP was higher than that in MGC-803 and MGC-803/DDP-KD, and the concentration of Cl- in KD group was the lowest (F=229, P<0.05). Conclusion The CLIC1 gene can induce the resistance of gastric cancer cell line MGC-803 to cisplatin in vitro. The mechanism may be related to the up-regulation of CLIC1 expression, which leads to an increase of intracellular chloride ion concentration. Key words: Gastric cancer; Chloride intracellular channel 1; Cisplatin; Drug resistance
{"title":"Establish cisplatin-resistant gastric cancer cell line MGC-803/cisplatin and reveal its mechanism","authors":"Jin-lu Liu, K. Zhao, Y. Qiu, Zhen Wang, Yuan-tian Mao, Bo-pei Li, Yeyang Chen","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.022","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.022","url":null,"abstract":"Objective \u0000To establish a cisplatin-resistant gastric cancer cell line MGC-803/cisplatin (DDP) and to explore its drug resistance mechanism. \u0000 \u0000 \u0000Methods \u0000The cisplatin was used to induce the drug resistance of MGC-803 cells. The half maximal inhibitory concentration (IC50) of the drug-resistant strains was detected by the cell counting kit-8 (CCK-8) method. The expression of chloride intracellular channel 1 (CLIC1) was detected by Western blotting analyses. The wild-type CLIC1 plasmid and the short hairpin RNA (shRNA) plasmid targeting CLIC1 were constructed. After Lipo3000 was transfected into gastric cancer cells, the IC50 of gastric cancer cells to cisplatin was detected, and then the intracellular Cl-concentration was detected by MQAE fluorescent probe. \u0000 \u0000 \u0000Results \u0000The cisplatin-resistant cell line MGC-803/DDP was successfully induced with an IC50 of (7.02±0.13) mg/L, while the IC50 of MGC803 was (1.29±0.09) mg/L, (t=36.090, P<0.05). The CLIC1 protein of MGC803/DDP was up-regulated by (2.27±0.15) times relative to MGC803 (t=7.841, P<0.05). After silencing the CLIC1 gene of MGC803/DDP, the IC50 of the CON, CN, and KD groups were (6.96±0.09), (6.93±0.15) and (3.02±0.20) mg/L, respectively, which was significantly decreased in the KD group (F=0.209, P<0.05). After overexpression of the CLIC1 gene in MGC803 cells, the IC50 of the CON, CN, and OE groups were (1.35±0.07), (1.25±0.07) and (4.77±0.12) mg/L, respectively, and the OE group was significantly elevated (F=0.508, P<0.05). The concentration of chloride ion was detected by MQAE. The relative fluorescence intensity of MGC-803, MGC-803/OE, MGC-803/DDP and MGC-803/DDP-KD were (1.02±0.03), (0.61±0.02), (0.67±0.01) and (1.39±0.02), respectively. The concentration of Cl- in MGC-803/OE cells was higher than that in MGC-803 (t=10.800, P<0.05), while the concentration of Cl- in MGC-803/DDP was higher than that in MGC-803 and MGC-803/DDP-KD, and the concentration of Cl- in KD group was the lowest (F=229, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The CLIC1 gene can induce the resistance of gastric cancer cell line MGC-803 to cisplatin in vitro. The mechanism may be related to the up-regulation of CLIC1 expression, which leads to an increase of intracellular chloride ion concentration. \u0000 \u0000 \u0000Key words: \u0000Gastric cancer; Chloride intracellular channel 1; Cisplatin; Drug resistance","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2200-2202"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43136441","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.017
Hongwei Xu, Yutian Luo, Dajun Wang, Liang Wang
Objective To investigate the expression of long chain non coding RNA (lncRNA) gastric cancer-associated transcript 3 (GACAT3) in hepatocellular carcinoma and its effect on proliferation, invasion and migration of HepG2 cells and its potential mechanism. Methods real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the differential expression of lncRNA GACAT3 in liver cancer tissue, adjacent tissues, PLC/PRF/5, Hep3B, HepG2 cell lines and normal liver cells THLE-3. Negative control small interfering RNA (siRNA) and lncRNA GACAT3 siRNA were transfected into HepG2 cells, after 48 h, Real-time PCR was used to detect the expression of lncRNA GACAT3 in transfected cells. Methyl thiazol tetrazolium (MTT) assay was used to detect cell proliferation in each group at 24, 48 and 72 h. The cell migration and invasion were detected by cell scratches experiments and Transwell assay, and the expression of Wnt/β-catenin signaling pathway protein was detected by Western blotting. Results The expression of lncRNA GACAT3 in hepatocellular carcinoma tissues (3.10±1.93) was significantly higher than that in adjacent tissues (1.02±0.59, t=8.086, P<0.01); The expression of lncRNA GACAT3 in PLC/PRF/5 (1.92±0.21, t=7.074, P<0.01), Hep3B (2.42±0.25, t=9.327, P<0.01) and HepG2 cells(2.96±0.31, t=10.582, P<0.01) were significantly higher than that in normal liver cells THLE-3 (0.97±0.10). Compared with the silencing control group (1.03±0.11), the expression of lncRNA GACAT3 in the GACAT3 group (0.16±0.02) was significantly decreased (t=13.478, P<0.01). Cell proliferation was significantly reduced at 48 h and 72 h (P<0.01). Scratch healing rate and migratory and invasive ability were significantly decreased (P<0.01). The expression of Wnt/β-catenin signaling pathway protein β-catenin (0.15±0.02) and its downstream proteins c-Myc (0.22±0.02) and Cyclin D1 (0.21±0.02) were significantly lower than that of the silencing control group(0.81±0.08, t=13.863; 0.65±0.07, t=9.731; 0.78±0.08, t=10.230; P all<0.01). The expression of p-β-catenin protein (0.75±0.08) was significantly higher than that of the silencing control group (0.27±0.03, t=11.972, P<0.01). Conclusion LncRNA GACAT3 is highly expressed in hepatocellular carcinoma. Silencing LncRNA GACAT3 can inhibit the proliferation, migration and invasion of hepatocellular carcinoma cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway. Key words: Long chain non coding RNA gastric cancer-associated transcript 3; Hepatocellular carcinoma; HepG2; Wnt/β-catenin signaling pathway
{"title":"Expression of long chain non coding RNA gastric cancer-associated transcript 3 in hepatocellular carcinoma and its effect on biological characteristics of HepG2 cells","authors":"Hongwei Xu, Yutian Luo, Dajun Wang, Liang Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.017","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.017","url":null,"abstract":"Objective \u0000To investigate the expression of long chain non coding RNA (lncRNA) gastric cancer-associated transcript 3 (GACAT3) in hepatocellular carcinoma and its effect on proliferation, invasion and migration of HepG2 cells and its potential mechanism. \u0000 \u0000 \u0000Methods \u0000real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the differential expression of lncRNA GACAT3 in liver cancer tissue, adjacent tissues, PLC/PRF/5, Hep3B, HepG2 cell lines and normal liver cells THLE-3. Negative control small interfering RNA (siRNA) and lncRNA GACAT3 siRNA were transfected into HepG2 cells, after 48 h, Real-time PCR was used to detect the expression of lncRNA GACAT3 in transfected cells. Methyl thiazol tetrazolium (MTT) assay was used to detect cell proliferation in each group at 24, 48 and 72 h. The cell migration and invasion were detected by cell scratches experiments and Transwell assay, and the expression of Wnt/β-catenin signaling pathway protein was detected by Western blotting. \u0000 \u0000 \u0000Results \u0000The expression of lncRNA GACAT3 in hepatocellular carcinoma tissues (3.10±1.93) was significantly higher than that in adjacent tissues (1.02±0.59, t=8.086, P<0.01); The expression of lncRNA GACAT3 in PLC/PRF/5 (1.92±0.21, t=7.074, P<0.01), Hep3B (2.42±0.25, t=9.327, P<0.01) and HepG2 cells(2.96±0.31, t=10.582, P<0.01) were significantly higher than that in normal liver cells THLE-3 (0.97±0.10). Compared with the silencing control group (1.03±0.11), the expression of lncRNA GACAT3 in the GACAT3 group (0.16±0.02) was significantly decreased (t=13.478, P<0.01). Cell proliferation was significantly reduced at 48 h and 72 h (P<0.01). Scratch healing rate and migratory and invasive ability were significantly decreased (P<0.01). The expression of Wnt/β-catenin signaling pathway protein β-catenin (0.15±0.02) and its downstream proteins c-Myc (0.22±0.02) and Cyclin D1 (0.21±0.02) were significantly lower than that of the silencing control group(0.81±0.08, t=13.863; 0.65±0.07, t=9.731; 0.78±0.08, t=10.230; P all<0.01). The expression of p-β-catenin protein (0.75±0.08) was significantly higher than that of the silencing control group (0.27±0.03, t=11.972, P<0.01). \u0000 \u0000 \u0000Conclusion \u0000LncRNA GACAT3 is highly expressed in hepatocellular carcinoma. Silencing LncRNA GACAT3 can inhibit the proliferation, migration and invasion of hepatocellular carcinoma cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway. \u0000 \u0000 \u0000Key words: \u0000Long chain non coding RNA gastric cancer-associated transcript 3; Hepatocellular carcinoma; HepG2; Wnt/β-catenin signaling pathway","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2183-2186"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49219681","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.006
Qian Cheng, Dafei Chai, Gang Wang, Zheng Lu, J. Bai
Objective To explore the effect of inhibition of poly (ADP-ribose) polymerase-1 (PARP-1) (AG14361) on the sensitivity of cells to arsenic trioxide (As2O3) in treating renal cancer cells. Methods Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, colony formation assay, single cell gel electrophoresis and micronucleus assay, flow cytometry were used to detect the cytotoxicity, genotoxicity and apoptosis of As2O3 combined with AG14361 on renal cancer cells. Results The colony formation rate [Combination group vs. As2O3 group: OS-RC-2, (43.23±3.12)% vs. (62.35±2.69)%; 786-O, (36.23±3.12)% vs. (58.31±2.42)%; HK-2, (56.32±3.46)% vs. (68.92±3.12)%], The difference was statistically significant (P<0.05); Single cell gel electrophoresis and micronucleus test showed that DNA and chromosome damage increased in the combination group compared with As2O3 group, and the difference was statistically significant (P<0.05). Flow cytometry shows, the combination group significantly enhanced the apoptotic rate of renal cancer cells compared with the As2O3 group [OS-RC-2, (59.90±86.75)% vs. (17.00±3.61)%; 786-O, (41.47±4.11)% vs. (15.17±1.99)%; HK-2, (30.43±4.92)% vs. (13.17±3.26)%]. Conclusion AG14361 has a sensitizing effect on the killing effect of As2O3-induced renal cancer cells, and its mechanism may be through inhibition of single-double-strand damage repair, which in turn causes chromosome damage. Key words: Renal cancer cells; As2O3; AG14361; Sensitizing
{"title":"Effect of polyadenyl diphosphate ribose polymerase 1 inhibitor on the sensitivity of As2O3 in treating renal cancer cells in vitro study","authors":"Qian Cheng, Dafei Chai, Gang Wang, Zheng Lu, J. Bai","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.006","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.006","url":null,"abstract":"Objective \u0000To explore the effect of inhibition of poly (ADP-ribose) polymerase-1 (PARP-1) (AG14361) on the sensitivity of cells to arsenic trioxide (As2O3) in treating renal cancer cells. \u0000 \u0000 \u0000Methods \u0000Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, colony formation assay, single cell gel electrophoresis and micronucleus assay, flow cytometry were used to detect the cytotoxicity, genotoxicity and apoptosis of As2O3 combined with AG14361 on renal cancer cells. \u0000 \u0000 \u0000Results \u0000The colony formation rate [Combination group vs. As2O3 group: OS-RC-2, (43.23±3.12)% vs. (62.35±2.69)%; 786-O, (36.23±3.12)% vs. (58.31±2.42)%; HK-2, (56.32±3.46)% vs. (68.92±3.12)%], The difference was statistically significant (P<0.05); Single cell gel electrophoresis and micronucleus test showed that DNA and chromosome damage increased in the combination group compared with As2O3 group, and the difference was statistically significant (P<0.05). Flow cytometry shows, the combination group significantly enhanced the apoptotic rate of renal cancer cells compared with the As2O3 group [OS-RC-2, (59.90±86.75)% vs. (17.00±3.61)%; 786-O, (41.47±4.11)% vs. (15.17±1.99)%; HK-2, (30.43±4.92)% vs. (13.17±3.26)%]. \u0000 \u0000 \u0000Conclusion \u0000AG14361 has a sensitizing effect on the killing effect of As2O3-induced renal cancer cells, and its mechanism may be through inhibition of single-double-strand damage repair, which in turn causes chromosome damage. \u0000 \u0000 \u0000Key words: \u0000Renal cancer cells; As2O3; AG14361; Sensitizing","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2148-2150"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49648069","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.004
Xu Hailiang, Gu Chaohui
Objective To investigate the expression and clinical significance of Cysteine-rich protein 61 (Cyr61) protein in prostate cancer. Methods We selected 68 cases of the prostate cancer tissues and 60 cases of benign prostatic hyperplasia tissues in our hospital. Immunohistochemistry was used to detect the expression of Cyr61 protein in prostate cancer tissues and benign prostatic hyperplasia tissues, and then the difference of expression between the two groups and its relationship with pathological parameters of prostate cancer were analyzed. Results The expression of Cyr61 protein in prostate cancer tissues was significantly higher than that in benign prostatic hyperplasia tissues (85.3% vs. 8.3%, χ2=75.536, P 0.05), TNM staging (χ2=0.342, P>0.05), regional lymph node metastasis (χ2=0.176, P>0.05) and distant metastasis (χ2=0.673, P>0.05), but was significantly correlated with preoperative total prostate specific antigen (tPSA) level (χ2=8.492, P<0.05), differentiation degree (χ2=13.028, P<0.05) and Gleason scores (χ2=4.032, P<0.05). Conclusion The expression of Cyr61 protein in prostate cancer tissues were significantly up-regulated, which helps to assess the progress of prostate cancer. Key words: Cysteine-rich protein 61 protein; Prostate carcinoma; Expression
{"title":"Expression of cysteine-rich protein 61 protein in prostate carcinoma and their clinical significance","authors":"Xu Hailiang, Gu Chaohui","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.004","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.004","url":null,"abstract":"Objective \u0000To investigate the expression and clinical significance of Cysteine-rich protein 61 (Cyr61) protein in prostate cancer. \u0000 \u0000 \u0000Methods \u0000We selected 68 cases of the prostate cancer tissues and 60 cases of benign prostatic hyperplasia tissues in our hospital. Immunohistochemistry was used to detect the expression of Cyr61 protein in prostate cancer tissues and benign prostatic hyperplasia tissues, and then the difference of expression between the two groups and its relationship with pathological parameters of prostate cancer were analyzed. \u0000 \u0000 \u0000Results \u0000The expression of Cyr61 protein in prostate cancer tissues was significantly higher than that in benign prostatic hyperplasia tissues (85.3% vs. 8.3%, χ2=75.536, P 0.05), TNM staging (χ2=0.342, P>0.05), regional lymph node metastasis (χ2=0.176, P>0.05) and distant metastasis (χ2=0.673, P>0.05), but was significantly correlated with preoperative total prostate specific antigen (tPSA) level (χ2=8.492, P<0.05), differentiation degree (χ2=13.028, P<0.05) and Gleason scores (χ2=4.032, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The expression of Cyr61 protein in prostate cancer tissues were significantly up-regulated, which helps to assess the progress of prostate cancer. \u0000 \u0000 \u0000Key words: \u0000Cysteine-rich protein 61 protein; Prostate carcinoma; Expression","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2141-2143"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46403198","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.010
Xuhui Zhang, Fan Liu, Qiqi Qin, Xiaobin Yuan, Min Zhang
Objective The purpose of this study was to investigate the factors influencing the thermal effects of holmium laser lithotripsy of urinary calculi and its safety. Methods We constructed an in vitro model of holmium laser thermal effect, then studied the affected factors about thermal effects of holmium laser lithotripsy: different power and energy patterns (1.0 J×10 hz, 0.5 J×20 hz, 1.0 J×20 hz, 2.0 J×20 hz), the perfusion fluid temperature (20 ℃ physiological saline, 37 ℃ physiological saline), the infusion speed (0 ml/min, 10 ml/min, 16 ml/min), optical fiber diameter (200, 365, 550 μm dahua holmium laser optical fiber), gravel model (gravel group and empty group). The holmium laser stimulated 120 s, during which the electronic thermometer measured and recorded temperature inside and outside the model each second. Each group experiments were repeated 3 times, and the average value of the three groups of experimental data was taken as the final experimental result. Independent sample T test or One-way ANOVA was used for comparison between groups. The optimal scale regression method was used to construct the influencing factor model of holmium laser thermal effect, and P<0.05 was considered as statistically significant difference. Results As the test begins, the temperature in the test tube increased rapidly, at around the 40 s, the temperature reached the plateau stage, and then the temperature tend to be stable. The results of optimal scale regression analysis showed that power and energy mode (F=18.533, P<0.05), perfusion velocity (F=220.461, P<0.01; F=495.783, P<0.01; F=32.287, P<0.01; F=429.237, P<0.01), perfusion liquid temperature (t=-4.657, P<0.01; t=-4.91, P<0.01; t=-4.157, P<0.05; t=-2.933, P<0.05) and fiber diameter (F=15.88, P<0.01; F=137.053, P<0.01; F=413.411, P<0.01; F=106.437, P<0.01) were the influencing factors of the thermal effect of holmium laser, and the difference was statistically significant (F=228.857, P<0.01). Conclusion The power and energy of holmium laser, perfusion rate, perfusion temperature and fiber diameter were the influencing factors of the thermal effect of holmium laser. Reducing the power of holmium laser, increasing the perfusion speed, using fiber with small diameter and choosing normal temperature perfusion solution were all helpful to reduce the thermal effects of holmium laser. Key words: Holmium laser; Thermal effect; Model in vitro
{"title":"The thermal effects of holmium laser lithotripsy in urinary calculus in an vitro model","authors":"Xuhui Zhang, Fan Liu, Qiqi Qin, Xiaobin Yuan, Min Zhang","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.010","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.010","url":null,"abstract":"Objective \u0000The purpose of this study was to investigate the factors influencing the thermal effects of holmium laser lithotripsy of urinary calculi and its safety. \u0000 \u0000 \u0000Methods \u0000We constructed an in vitro model of holmium laser thermal effect, then studied the affected factors about thermal effects of holmium laser lithotripsy: different power and energy patterns (1.0 J×10 hz, 0.5 J×20 hz, 1.0 J×20 hz, 2.0 J×20 hz), the perfusion fluid temperature (20 ℃ physiological saline, 37 ℃ physiological saline), the infusion speed (0 ml/min, 10 ml/min, 16 ml/min), optical fiber diameter (200, 365, 550 μm dahua holmium laser optical fiber), gravel model (gravel group and empty group). The holmium laser stimulated 120 s, during which the electronic thermometer measured and recorded temperature inside and outside the model each second. Each group experiments were repeated 3 times, and the average value of the three groups of experimental data was taken as the final experimental result. Independent sample T test or One-way ANOVA was used for comparison between groups. The optimal scale regression method was used to construct the influencing factor model of holmium laser thermal effect, and P<0.05 was considered as statistically significant difference. \u0000 \u0000 \u0000Results \u0000As the test begins, the temperature in the test tube increased rapidly, at around the 40 s, the temperature reached the plateau stage, and then the temperature tend to be stable. The results of optimal scale regression analysis showed that power and energy mode (F=18.533, P<0.05), perfusion velocity (F=220.461, P<0.01; F=495.783, P<0.01; F=32.287, P<0.01; F=429.237, P<0.01), perfusion liquid temperature (t=-4.657, P<0.01; t=-4.91, P<0.01; t=-4.157, P<0.05; t=-2.933, P<0.05) and fiber diameter (F=15.88, P<0.01; F=137.053, P<0.01; F=413.411, P<0.01; F=106.437, P<0.01) were the influencing factors of the thermal effect of holmium laser, and the difference was statistically significant (F=228.857, P<0.01). \u0000 \u0000 \u0000Conclusion \u0000The power and energy of holmium laser, perfusion rate, perfusion temperature and fiber diameter were the influencing factors of the thermal effect of holmium laser. Reducing the power of holmium laser, increasing the perfusion speed, using fiber with small diameter and choosing normal temperature perfusion solution were all helpful to reduce the thermal effects of holmium laser. \u0000 \u0000 \u0000Key words: \u0000Holmium laser; Thermal effect; Model in vitro","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2160-2162"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43989857","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.012
W. Di, H. Ji, Qinghuai Li
Objective To investigate the regulating effects of microRNA (miRNA, miR)-122 on the expression of ilent information regulator 1 (SIRT1), and the proliferation abilities of papillary thyroid cancer cells. Methods Real-time quantitative polymerase chain reaction (Real-time PCR) was used to observe the expression of miR-122 in papillary thyroid carcinoma and adjacent tissues and to analyze its correlation with SIRT1 expression. After transfecting miR-122 mimics into human papillary thyroid carcinoma cell line TPC-1, the dual luciferase reporter gene assay was used to observe the regulation of miR-122 on SIRT1 gene. The effect of miR-122 on the proliferation of thyroid cancer cells was observed by CCK-8 assay. At the same time, the overexpressed SIRT1 plasmid was co-transfected into TPC-1 cells, and then the effect of miR-122 and SIRT1 expression on the proliferation of papillary thyroid carcinoma cells was detected by CCK-8 assay. Analysis was performed using GraphPad Prism 7.0 statistical software. Correlation analysis was performed using Spearman rank correlation analysis. Student’s t test was used to compare the two sample means. Results Compared with adjacent tissues (1.20±0.15), the expression level of miR-122 (3.06±0.26) in papillary thyroid carcinoma was significantly down-regulated, and the difference was statistically significant (t=5.972, P<0.01). Correlation analysis showed that miR-122 was negatively correlated with SIRT1 mRNA expression (r=-0.621, P<0.01). The relative luciferase activity of SIRT1 in miR-122 mimics transfection group was significantly lower than that in the control group, and the difference was statistically significant (t=17.730, P<0.01). The cell proliferation ability of miR-122 mimics group was significantly lower than that of the control group, and the difference was statistically significant (t=3.991, P<0.01). After co-transfection of the SIRT1 overexpression plasmid, the inhibition of the proliferation of thyroid papillary carcinoma cells by miR-122 mimics was reversed. Conclusion miR-122 can inhibit the proliferation of papillary thyroid cancer cells, and its mechanism may be related to the targeted inhibition of SIRT1 expression. Key words: MicroRNA-122; Ilent information regulator 1; Papillary thyroid cancer; Proliferation
{"title":"MicroRNA-92b suppresses proliferation of papillary thyroid cancer cells by regulating ilent information regulator 1 gene expression","authors":"W. Di, H. Ji, Qinghuai Li","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.012","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.012","url":null,"abstract":"Objective \u0000To investigate the regulating effects of microRNA (miRNA, miR)-122 on the expression of ilent information regulator 1 (SIRT1), and the proliferation abilities of papillary thyroid cancer cells. \u0000 \u0000 \u0000Methods \u0000Real-time quantitative polymerase chain reaction (Real-time PCR) was used to observe the expression of miR-122 in papillary thyroid carcinoma and adjacent tissues and to analyze its correlation with SIRT1 expression. After transfecting miR-122 mimics into human papillary thyroid carcinoma cell line TPC-1, the dual luciferase reporter gene assay was used to observe the regulation of miR-122 on SIRT1 gene. The effect of miR-122 on the proliferation of thyroid cancer cells was observed by CCK-8 assay. At the same time, the overexpressed SIRT1 plasmid was co-transfected into TPC-1 cells, and then the effect of miR-122 and SIRT1 expression on the proliferation of papillary thyroid carcinoma cells was detected by CCK-8 assay. Analysis was performed using GraphPad Prism 7.0 statistical software. Correlation analysis was performed using Spearman rank correlation analysis. Student’s t test was used to compare the two sample means. \u0000 \u0000 \u0000Results \u0000Compared with adjacent tissues (1.20±0.15), the expression level of miR-122 (3.06±0.26) in papillary thyroid carcinoma was significantly down-regulated, and the difference was statistically significant (t=5.972, P<0.01). Correlation analysis showed that miR-122 was negatively correlated with SIRT1 mRNA expression (r=-0.621, P<0.01). The relative luciferase activity of SIRT1 in miR-122 mimics transfection group was significantly lower than that in the control group, and the difference was statistically significant (t=17.730, P<0.01). The cell proliferation ability of miR-122 mimics group was significantly lower than that of the control group, and the difference was statistically significant (t=3.991, P<0.01). After co-transfection of the SIRT1 overexpression plasmid, the inhibition of the proliferation of thyroid papillary carcinoma cells by miR-122 mimics was reversed. \u0000 \u0000 \u0000Conclusion \u0000miR-122 can inhibit the proliferation of papillary thyroid cancer cells, and its mechanism may be related to the targeted inhibition of SIRT1 expression. \u0000 \u0000 \u0000Key words: \u0000MicroRNA-122; Ilent information regulator 1; Papillary thyroid cancer; Proliferation","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2167-2169"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42269186","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 : 2019-12-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2019.12.021
P. Yu, Q. Fu, Hao Zhang, Shu-Yi Zhao, Pan Liu, M. Hu
Objective To investigate the expression of S-phase kinase-associated protein 1 (SKP1) in intrahepatic cholangiocarcinoma and to investigate its effects on proliferation, migration and invasion of intrahepatic cholangiocarcinoma. Methods Western blotting assay was used to determine the expression difference of SKP1 in intrahepatic cholangiocarcinoma and normal tissues. The SKP1 interference vector and empty vector were constructed and stably transfected into intrahepatic cholangiocarcinoma cells by lentivirus packaging to form an experimental group and a control group. The expression of SKP1 was detected by immunoblotting assay. The proliferation of experimental group and control group after transfection was detected by cell counting kit-8 (CCK-8) assay. The migration ability was detected by scratch test. The invasion ability of Transwell was detected by Transwellassay. Results The expression of SKP1 in intrahepatic cholangiocarcinoma was (1.701±0.215) times, and the difference was statistically significant (t=8.512, P<0.05). Lentiviral transfection of RBE cells decreased the expression of SKP1, and the Western blotting assay decreased the experimental group by (3.632±0.335) times, and the difference was statistically significant (t=11.376, P<0.01). The CCK-8 experiment showed that the relative survival rates of the experimental group at 24, 48, and 72 hours were (53.136±0.314)%, (42.661±0.248)% and (35.037±0.203)%, respectively, and the differences were statistically significant (t=8.594, 8.899, 9.210, P<0.01). The scratch test showed that the number of migrated cells in the experimental group at 24 and 48 hours (41.198±3.092) and (64.773±6.437) were significantly lower than those in the control group (113.487±3.514) and (168.767±8.151), and the difference was statistically significant (t=5.430, 6.224, P<0.05). The Transwell experiment showed that the number of perforated cells in the experimental group was (34.185±3.124), which was significantly lower than that in the control group (121.234±6.814), and the difference was statistically significant (t=9.918, P<0.05). Conclusion The expression of SKP1 in intrahepatic cholangiocarcinoma is higher than that in normal tissues. Decreasing SKP1 expression can inhibit the proliferation, migration and invasion of intrahepatic cholangiocarcinoma. Key words: S phase kinase associated protein-1; Intrahepatic cholangiocarcinoma; Ubiquitination; Proliferation; Migrate; Invasion
{"title":"S phase kinase-associated protein 1 can inhibit the proliferation and invasion of intrahepatic cholangiocarcinoma","authors":"P. Yu, Q. Fu, Hao Zhang, Shu-Yi Zhao, Pan Liu, M. Hu","doi":"10.3760/CMA.J.ISSN.1001-9030.2019.12.021","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2019.12.021","url":null,"abstract":"Objective \u0000To investigate the expression of S-phase kinase-associated protein 1 (SKP1) in intrahepatic cholangiocarcinoma and to investigate its effects on proliferation, migration and invasion of intrahepatic cholangiocarcinoma. \u0000 \u0000 \u0000Methods \u0000Western blotting assay was used to determine the expression difference of SKP1 in intrahepatic cholangiocarcinoma and normal tissues. The SKP1 interference vector and empty vector were constructed and stably transfected into intrahepatic cholangiocarcinoma cells by lentivirus packaging to form an experimental group and a control group. The expression of SKP1 was detected by immunoblotting assay. The proliferation of experimental group and control group after transfection was detected by cell counting kit-8 (CCK-8) assay. The migration ability was detected by scratch test. The invasion ability of Transwell was detected by Transwellassay. \u0000 \u0000 \u0000Results \u0000The expression of SKP1 in intrahepatic cholangiocarcinoma was (1.701±0.215) times, and the difference was statistically significant (t=8.512, P<0.05). Lentiviral transfection of RBE cells decreased the expression of SKP1, and the Western blotting assay decreased the experimental group by (3.632±0.335) times, and the difference was statistically significant (t=11.376, P<0.01). The CCK-8 experiment showed that the relative survival rates of the experimental group at 24, 48, and 72 hours were (53.136±0.314)%, (42.661±0.248)% and (35.037±0.203)%, respectively, and the differences were statistically significant (t=8.594, 8.899, 9.210, P<0.01). The scratch test showed that the number of migrated cells in the experimental group at 24 and 48 hours (41.198±3.092) and (64.773±6.437) were significantly lower than those in the control group (113.487±3.514) and (168.767±8.151), and the difference was statistically significant (t=5.430, 6.224, P<0.05). The Transwell experiment showed that the number of perforated cells in the experimental group was (34.185±3.124), which was significantly lower than that in the control group (121.234±6.814), and the difference was statistically significant (t=9.918, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The expression of SKP1 in intrahepatic cholangiocarcinoma is higher than that in normal tissues. Decreasing SKP1 expression can inhibit the proliferation, migration and invasion of intrahepatic cholangiocarcinoma. \u0000 \u0000 \u0000Key words: \u0000S phase kinase associated protein-1; Intrahepatic cholangiocarcinoma; Ubiquitination; Proliferation; Migrate; Invasion","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":"2197-2199"},"PeriodicalIF":0.0,"publicationDate":"2019-12-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48042980","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}
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