Objective To investigate the effects of rosuvastatin on proliferation, osteogenic differentiation and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) expression of mesenchymal stem cells (MSCs). Methods MSCs were isolated. Rosuvastatin at concentrations of 1×10-11, 1×10-9 and 1×10-7 mol/L were added to the experimental groups, and dimethylsurfoxide (DMSO) served as control. Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay. After 3, 7, 14 and 21 days of osteogenic induction, alkaline phosphatase (ALP) quantitative analysis, alizarin red staining and quantitative analysis, real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were performed. T-test was used for comparison between two groups, and ANOVA for more than two groups. Results At 24 h and 48 h, the proliferation of MSCs in high concentration group was significantly higher than in the control group (24 h: 3.57±0.24 vs. 3.14±0.14, t=-3.851, P<0.05; 48 h: 4.19±0.18 vs. 3.44±0.11, t=-6.780, P<0.05). Until 72 h, the proliferation with 1×10-9 mol/L rosuvastatin was also significantly increased (5.42±0.13 vs. 4.47±0.16, t=-8.700, P<0.05). At 3rd day after induction culture, 1×10-7 mol/L rosuvastatin significantly enhanced the genes expression of BMP2, Runx 2, OPN 2, OPN and ColⅠ (BMP2: 2.1±0.2 vs. 1.0±0.2, t=-6.736, P<0.05; Runx2: 5.2±0.6 vs. 1.0±0.1, t=-11.959, P<0.05; OPN: 1.8±0.4 vs. 1.0±0.2, t=-3.098, P<0.05; ColⅠ: 1.9±0.3 vs. 1.0±0.3, t=-3.674, P<0.05); 1×10-7 mol/L rosuvastatin could increase the expression of p-Akt/PI3K. The ALP activity after adding 1×10-9 or 1×10-7 mol/L rosuvastatin was significantly increased at 14th and 21st day (14 d: 12.07±1.67, 18.32±2.26 vs. 3.43±0.36, F=7.200, P<0.05; 21 d: 10.74±1.27, 14.71±3.18 vs. 5.76±0.63, F=6.489, P<0.05). The quantitative results of alizarin red were similar to those of the staining. Meanwhile, the quantitative values of alizarin red in the 1×10-9 and 1×10-7 mol/L groups were also significantly increased (14 d: 5.6±0.8, 6.2±1.2 vs. 1.0±0.2, F=5.600, P<0.05; 21 d: 5.8±1.1, 7.1±1.8 vs. 2.4±0.3, F=5.956, P<0.05). Conclusion Rosuvastatin promotes proliferation and osteogenic differentiation of MSCs, which may be related to PI3K/Akt signaling. Key words: Rosuvastatin; Phosphatidylinositol-3-kinase/protein kinase B signaling pathway; Mesenchymal stem cells; Osteogenic differentiation
目的探讨瑞舒伐他汀对间充质干细胞(MSCs)增殖、成骨分化及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B (Akt)表达的影响。方法分离MSCs。实验组分别加入浓度为1×10-11、1×10-9和1×10-7 mol/L的瑞舒伐他汀，对照组分别加入二甲苏福啶(DMSO)。采用甲基噻唑四氮唑(MTT)法检测细胞增殖。成骨诱导3、7、14、21 d后进行碱性磷酸酶(ALP)定量分析、茜素红染色及定量分析、实时定量聚合酶链反应(real-time PCR)和Western blotting。两组间比较采用t检验，两组以上采用方差分析。结果24 h、48 h，高浓度组MSCs的增殖明显高于对照组(24 h: 3.57±0.24 vs. 3.14±0.14,t=-3.851, P<0.05;48 h: 4.19±0.18和3.44±0.11,t = -6.780, P < 0.05)。1×10-9 mol/L瑞舒伐他汀治疗至72 h时，细胞增殖也显著增加(5.42±0.13∶4.47±0.16,t=-8.700, P<0.05)。诱导培养后第3天，1×10-7 mol/L瑞舒伐他汀显著提高BMP2、runx2、opn2、OPN和Col基因表达Ⅰ(BMP2: 2.1±0.2 vs. 1.0±0.2,t=-6.736, P<0.05;Runx2: 5.2±0.6 vs. 1.0±0.1,t=-11.959, P<0.05;OPN: 1.8±0.4 vs. 1.0±0.2,t=-3.098, P<0.05;ColⅠ:1.9±0.3 vs. 1.0±0.3,t=-3.674, P<0.05);1×10-7 mol/L瑞舒伐他汀可增加p-Akt/PI3K的表达。添加1×10-9或1×10-7 mol/L瑞舒伐他汀后第14天和第21天ALP活性显著升高(14 d: 12.07±1.67,18.32±2.26 vs. 3.43±0.36,F=7.200, P<0.05;21 d: 10.74±1.27,14.71±3.18和5.76±0.63,F = 6.489, P < 0.05)。茜素红的定量结果与染色结果相似。同时，1×10-9和1×10-7 mol/L组的芹素红定量值也显著升高(14 d: 5.6±0.8、6.2±1.2 vs. 1.0±0.2,F=5.600, P<0.05;21 d: 5.8±1.1,7.1±1.8和2.4±0.3,F = 5.956, P < 0.05)。结论瑞舒伐他汀促进MSCs增殖和成骨分化，可能与PI3K/Akt信号通路有关。关键词:瑞舒伐他汀;磷脂酰肌醇-3激酶/蛋白激酶B信号通路;间充质干细胞;成骨分化
{"title":"Rosuvastatin regulates osteogenic differentiation of mesenchymal stem cells through phosphatidylinositol-3-kinase/protein kinase B","authors":"Xuepeng Wang, Chunchun Zou, Changju Hou, Maoqiang Li, Z. Bian, Liulong Zhu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.029","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.029","url":null,"abstract":"Objective \u0000To investigate the effects of rosuvastatin on proliferation, osteogenic differentiation and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) expression of mesenchymal stem cells (MSCs). \u0000 \u0000 \u0000Methods \u0000MSCs were isolated. Rosuvastatin at concentrations of 1×10-11, 1×10-9 and 1×10-7 mol/L were added to the experimental groups, and dimethylsurfoxide (DMSO) served as control. Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay. After 3, 7, 14 and 21 days of osteogenic induction, alkaline phosphatase (ALP) quantitative analysis, alizarin red staining and quantitative analysis, real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were performed. T-test was used for comparison between two groups, and ANOVA for more than two groups. \u0000 \u0000 \u0000Results \u0000At 24 h and 48 h, the proliferation of MSCs in high concentration group was significantly higher than in the control group (24 h: 3.57±0.24 vs. 3.14±0.14, t=-3.851, P<0.05; 48 h: 4.19±0.18 vs. 3.44±0.11, t=-6.780, P<0.05). Until 72 h, the proliferation with 1×10-9 mol/L rosuvastatin was also significantly increased (5.42±0.13 vs. 4.47±0.16, t=-8.700, P<0.05). At 3rd day after induction culture, 1×10-7 mol/L rosuvastatin significantly enhanced the genes expression of BMP2, Runx 2, OPN 2, OPN and ColⅠ (BMP2: 2.1±0.2 vs. 1.0±0.2, t=-6.736, P<0.05; Runx2: 5.2±0.6 vs. 1.0±0.1, t=-11.959, P<0.05; OPN: 1.8±0.4 vs. 1.0±0.2, t=-3.098, P<0.05; ColⅠ: 1.9±0.3 vs. 1.0±0.3, t=-3.674, P<0.05); 1×10-7 mol/L rosuvastatin could increase the expression of p-Akt/PI3K. The ALP activity after adding 1×10-9 or 1×10-7 mol/L rosuvastatin was significantly increased at 14th and 21st day (14 d: 12.07±1.67, 18.32±2.26 vs. 3.43±0.36, F=7.200, P<0.05; 21 d: 10.74±1.27, 14.71±3.18 vs. 5.76±0.63, F=6.489, P<0.05). The quantitative results of alizarin red were similar to those of the staining. Meanwhile, the quantitative values of alizarin red in the 1×10-9 and 1×10-7 mol/L groups were also significantly increased (14 d: 5.6±0.8, 6.2±1.2 vs. 1.0±0.2, F=5.600, P<0.05; 21 d: 5.8±1.1, 7.1±1.8 vs. 2.4±0.3, F=5.956, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Rosuvastatin promotes proliferation and osteogenic differentiation of MSCs, which may be related to PI3K/Akt signaling. \u0000 \u0000 \u0000Key words: \u0000Rosuvastatin; Phosphatidylinositol-3-kinase/protein kinase B signaling pathway; Mesenchymal stem cells; Osteogenic differentiation","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"101-104"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46317928","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.1001-9030.2020.01.026
Jing Qin, Chen Zhang, Weiping Tian, J. Chen, Lin Fang
Objective To investigate the effect of chimeric promoter composed of hypoxia response element (HRE) and tumor specific cell proliferation-associated nuclear antigen (Ki-67) promoter on the replication ability of oncolytic adenovirus and its cytotoxic activity under hypoxic conditions. Methods The HRE sequence, as an enhancer, was inserted upstream of the Ki-67 promoter and, at the same time carried the reporter gene enhanced green fluorescent protein (EGFP) to construct oncolytic adenovirus Ad-HRE-Ki-67-EGFP, and the titer of adenoviruses was measured by the tissue culture infective dose (TCID50). Normal renal epithelial cells HK-2 and renal cancer cells OSRC-2 and ACHN were infected with two kinds of viruses under normoxic (21% O2) or hypoxic (1% O2) conditions respectively for 24 h, and the virus tumor-targeting was detected by fluorescence microscopy and flow cytometry. Western blotting was performed to detect the expression of hypoxia-related protein hypoxia-inducible factor-1 alpha (HIF-1α), vascular endothelial growth factor (VEGF) and the expression of E1A protein. Cell counting kit-8 (CCK-8) assay was used to detect the cytotoxity of virus on cells. Results The recombinant oncolytic virus Ad-HRE-Ki67-EGFP was successfully constructed. Under normoxic conditions, the efficiency of ACHN and OSRC-2 infection by chimeric HRE oncolytic adenovirus in renal cancer cells was (47.5±2.6)% and (35.2±3.4)%, while that in hypoxic conditions was (86.4±3.4)% and (73.5±2.3)%, indicating that the infection efficiency was higher under hypoxic conditions (ACHN: t=15.760; OSRC-2: t=16.190; P<0.01); Normal renal tubular epithelial cells HK-2 were infected with Ad-Ki-67-EGFP or Ad-HRE-Ki-67-EGFP under normoxic conditions. The infection efficiency was (22.4±3.1)% and (22.9±2.2)% respectively, compared with that of ACHN and OSRC-2 cells [ACHN: (68.4±3.3)% and (72.1±2.9)%; OSRC-2: (56.1±3.1)% and (61.4±2.3)%], the difference was statistically significant (Ad-Ki-67-EGFP: t=17.650, 13.390; Ad-HRE-Ki-67-EGFP: t=23.680, 21.430, P<0.01), indicating that the two adenoviruses have a targeting effect on renal cancer cells. Western blotting data showed that the expression of HIF-1α, VEGF and p53 protein were increased under hypoxic conditions and was time-dependent, indicating that hypoxia can regulate the transcription and translation of HIF-1α, VEGF and p53 proteins. Under hypoxic conditions, the expression level of E1A protein in OSRC-2 and ACHN infected with Ad-HRE-Ki67-EGFP was significantly higher than that after Ad-Ki67-EGFP infection. It indicated that HRE can enhance the replication ability of virus under hypoxic conditions. CCK-8 results showed that when the multiple infection (MOI) values were 20, 100, the survival rate of the Ad-HRE-Ki-67-EGFP-normoxia virus group in ACHN and OSRC-2 cells was (73.4±2.0)%, (56.4±1.5)% and (79.9±1.8)%, (61.3±2.7)%, the Ad-HRE-Ki-67-EGFP-hypoxia virus group was (63.1±2.0)%, (31.6±2.1)% and (68.1±2.6)%, (35.8±3.1)%, th
目的探讨缺氧反应元件(HRE)与肿瘤特异性细胞增殖相关核抗原(Ki-67)启动子嵌合启动子在缺氧条件下对溶瘤腺病毒复制能力及其细胞毒活性的影响。方法将HRE序列作为增强子插入Ki-67启动子上游，同时携带报告基因增强型绿色荧光蛋白(EGFP)构建溶瘤腺病毒Ad-HRE-Ki-67-EGFP，用组织培养感染剂量(TCID50)测定腺病毒滴度。将正常肾上皮细胞HK-2和肾癌细胞OSRC-2和ACHN分别在常氧(21% O2)和缺氧(1% O2)条件下感染24 h，用荧光显微镜和流式细胞术检测病毒的肿瘤靶向性。Western blotting检测缺氧相关蛋白-缺氧诱导因子-1α (HIF-1α)、血管内皮生长因子(VEGF)及E1A蛋白的表达。采用细胞计数试剂盒-8 (CCK-8)法检测病毒对细胞的毒性。结果成功构建了重组溶瘤病毒Ad-HRE-Ki67-EGFP。在常氧条件下，嵌合HRE溶瘤腺病毒感染肾癌细胞ACHN和OSRC-2的效率分别为(47.5±2.6)%和(35.2±3.4)%，而在缺氧条件下，ACHN和OSRC-2的感染效率分别为(86.4±3.4)%和(73.5±2.3)%，说明缺氧条件下ACHN和OSRC-2的感染效率更高(ACHN: t=15.760;OSRC-2: t = 16.190;P < 0.01);在正常条件下，用Ad-Ki-67-EGFP或Ad-HRE-Ki-67-EGFP感染正常肾小管上皮细胞HK-2。感染效率分别为(22.4±3.1)%和(22.9±2.2)%，而ACHN和OSRC-2细胞的感染效率分别为(68.4±3.3)%和(72.1±2.9)%;OSRC-2分别为(56.1±3.1)%和(61.4±2.3)%，差异有统计学意义(Ad-Ki-67-EGFP: t=17.650, 13.390;Ad-HRE-Ki-67-EGFP: t=23.680, 21.430, P<0.01)，说明两种腺病毒对肾癌细胞具有靶向作用。Western blotting数据显示，缺氧条件下HIF-1α、VEGF和p53蛋白表达增加，且具有时间依赖性，提示缺氧可调节HIF-1α、VEGF和p53蛋白的转录和翻译。缺氧条件下，感染Ad-HRE-Ki67-EGFP的OSRC-2和ACHN中E1A蛋白的表达水平显著高于感染Ad-Ki67-EGFP后的表达水平。说明HRE能增强病毒在缺氧条件下的复制能力。CCK-8结果显示，当多重感染(MOI)值为20、100时，ad - hre - ki -67- egfp -常氧病毒组在ACHN和ossc -2细胞中的存活率分别为(73.4±2.0)%、(56.4±1.5)%和(79.9±1.8)%、(61.3±2.7)%，ad - hre - ki -67- egfp -缺氧病毒组的存活率分别为(63.1±2.0)%、(31.6±2.1)%和(68.1±2.6)%、(35.8±3.1)%，ad - hre - ki -67- egfp -缺氧病毒组的增殖抑制作用较强[MOI (20): t=6.328、6.441;MOI (100): t=16.410, 10.790, P<0.01。结论在缺氧条件下，嵌合HRE序列重组溶瘤腺病毒在肾癌细胞中具有较强的复制能力和细胞杀伤作用。关键词:肾细胞癌;溶瘤腺病毒;缺氧反应元件;缺氧
{"title":"Antitumor effect of hypoxia response element chimeric tumor specific promoter regulated oncolytic adenovirus on renal cell carcinoma","authors":"Jing Qin, Chen Zhang, Weiping Tian, J. Chen, Lin Fang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.026","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.026","url":null,"abstract":"Objective \u0000To investigate the effect of chimeric promoter composed of hypoxia response element (HRE) and tumor specific cell proliferation-associated nuclear antigen (Ki-67) promoter on the replication ability of oncolytic adenovirus and its cytotoxic activity under hypoxic conditions. \u0000 \u0000 \u0000Methods \u0000The HRE sequence, as an enhancer, was inserted upstream of the Ki-67 promoter and, at the same time carried the reporter gene enhanced green fluorescent protein (EGFP) to construct oncolytic adenovirus Ad-HRE-Ki-67-EGFP, and the titer of adenoviruses was measured by the tissue culture infective dose (TCID50). Normal renal epithelial cells HK-2 and renal cancer cells OSRC-2 and ACHN were infected with two kinds of viruses under normoxic (21% O2) or hypoxic (1% O2) conditions respectively for 24 h, and the virus tumor-targeting was detected by fluorescence microscopy and flow cytometry. Western blotting was performed to detect the expression of hypoxia-related protein hypoxia-inducible factor-1 alpha (HIF-1α), vascular endothelial growth factor (VEGF) and the expression of E1A protein. Cell counting kit-8 (CCK-8) assay was used to detect the cytotoxity of virus on cells. \u0000 \u0000 \u0000Results \u0000The recombinant oncolytic virus Ad-HRE-Ki67-EGFP was successfully constructed. Under normoxic conditions, the efficiency of ACHN and OSRC-2 infection by chimeric HRE oncolytic adenovirus in renal cancer cells was (47.5±2.6)% and (35.2±3.4)%, while that in hypoxic conditions was (86.4±3.4)% and (73.5±2.3)%, indicating that the infection efficiency was higher under hypoxic conditions (ACHN: t=15.760; OSRC-2: t=16.190; P<0.01); Normal renal tubular epithelial cells HK-2 were infected with Ad-Ki-67-EGFP or Ad-HRE-Ki-67-EGFP under normoxic conditions. The infection efficiency was (22.4±3.1)% and (22.9±2.2)% respectively, compared with that of ACHN and OSRC-2 cells [ACHN: (68.4±3.3)% and (72.1±2.9)%; OSRC-2: (56.1±3.1)% and (61.4±2.3)%], the difference was statistically significant (Ad-Ki-67-EGFP: t=17.650, 13.390; Ad-HRE-Ki-67-EGFP: t=23.680, 21.430, P<0.01), indicating that the two adenoviruses have a targeting effect on renal cancer cells. Western blotting data showed that the expression of HIF-1α, VEGF and p53 protein were increased under hypoxic conditions and was time-dependent, indicating that hypoxia can regulate the transcription and translation of HIF-1α, VEGF and p53 proteins. Under hypoxic conditions, the expression level of E1A protein in OSRC-2 and ACHN infected with Ad-HRE-Ki67-EGFP was significantly higher than that after Ad-Ki67-EGFP infection. It indicated that HRE can enhance the replication ability of virus under hypoxic conditions. CCK-8 results showed that when the multiple infection (MOI) values were 20, 100, the survival rate of the Ad-HRE-Ki-67-EGFP-normoxia virus group in ACHN and OSRC-2 cells was (73.4±2.0)%, (56.4±1.5)% and (79.9±1.8)%, (61.3±2.7)%, the Ad-HRE-Ki-67-EGFP-hypoxia virus group was (63.1±2.0)%, (31.6±2.1)% and (68.1±2.6)%, (35.8±3.1)%, th","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"90-92"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43270531","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 observe the effects of long-chain non-coding RNA (lncRNA) HIF2PUT on the proliferation, migration and invasion of osteosarcoma cells. Methods Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of lncRNA HIF2PUT in 47 cases of osteosarcoma tissues (28 males and 19 females, the oldest was 45 years old, the youngest was 8 years old with an average of 20.24 years old) and 2 osteosarcoma cell lines (MG63 and HOS). After screening for stable transfectants by lentivirus, the methyl thiazol tetrazolium (MTT) assay and Transwell assay were applied to detect the effects of lncRNA HIF2PUT on proliferation, migration and invasion of osteosarcoma cells. According to the cell transfection, the experimental group (overexpressed lncRNA HIF2PUT group), the blank transfection group and the blank control group were respectively set. Data were expressed as mean±standard deviations (SD), and SPSS 21.0 software was used for statistical analysis. Results The expression of lncRNA HIF2PUT in 47 cases of osteosarcoma (39.56±0.48) was significantly lower than that in normal tissues (79.56±1.16, t=3.516, P<0.05). The expression level of lncRNA HIF2PUT in MG63 cells was significantly higher than that in HOS cells. Cell function experiments showed that lncRNA HIF2PUT had a significant effect on the proliferation, migration and invasion of osteosarcoma cells. Proliferation of MG63 and HOS cells was significantly inhibited by lncRNA HIF2PUT, and the proliferation index was significantly lower in lncRNA HIF2PUT group than in blank transfection group and blank control group (t=4.785, P<0.05). For the migration experiment, the migrating number of MG63 cells was (154.36±13.85), (196.32±13.52) and (258.63±12.35), respectively (t=3.968, P<0.01), and that of HOS cells was (131.24±10.58), (247.36±17.52) and (224.52±14.52) respectively (t=3.052, P<0.01). For the invasion experiment, the penetrating number of MG63 cells was (73.50±13.89), (98.63±12.57) and (112.30±15.29), respectively (t=23.178, P<0.01), and that of HOS cells was (70.74±13.49) (98.40±16.46) and 89.74±12.47, respectively (t=14.275, P<0.01). The migration and invasion ability of MG63 and HOS cells in lncRNA HIF2PUT group was lower than that in blank transfection group and blank control group. Conclusion The expression of lncRNA HIF2PUT was significantly low in osteosarcoma tissues and MG63 and HOS cell lines. The expression of lncRNA HIF2PUT significantly inhibited the proliferation, migration and invasion of osteosarcoma cells. Key words: Osteosarcoma; Long-chain non-coding RNA; Migration; Invasion
{"title":"Effects of long-chain non-coding HIF2PUT on migration and invasion of osteosarcoma cells","authors":"Caijuan Guo, Hui-bin Xu, Hong-jian Liu, Guofu Pi, Jianguang Sun, Yong Zhu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.049","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.049","url":null,"abstract":"Objective \u0000To observe the effects of long-chain non-coding RNA (lncRNA) HIF2PUT on the proliferation, migration and invasion of osteosarcoma cells. \u0000 \u0000 \u0000Methods \u0000Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of lncRNA HIF2PUT in 47 cases of osteosarcoma tissues (28 males and 19 females, the oldest was 45 years old, the youngest was 8 years old with an average of 20.24 years old) and 2 osteosarcoma cell lines (MG63 and HOS). After screening for stable transfectants by lentivirus, the methyl thiazol tetrazolium (MTT) assay and Transwell assay were applied to detect the effects of lncRNA HIF2PUT on proliferation, migration and invasion of osteosarcoma cells. According to the cell transfection, the experimental group (overexpressed lncRNA HIF2PUT group), the blank transfection group and the blank control group were respectively set. Data were expressed as mean±standard deviations (SD), and SPSS 21.0 software was used for statistical analysis. \u0000 \u0000 \u0000Results \u0000The expression of lncRNA HIF2PUT in 47 cases of osteosarcoma (39.56±0.48) was significantly lower than that in normal tissues (79.56±1.16, t=3.516, P<0.05). The expression level of lncRNA HIF2PUT in MG63 cells was significantly higher than that in HOS cells. Cell function experiments showed that lncRNA HIF2PUT had a significant effect on the proliferation, migration and invasion of osteosarcoma cells. Proliferation of MG63 and HOS cells was significantly inhibited by lncRNA HIF2PUT, and the proliferation index was significantly lower in lncRNA HIF2PUT group than in blank transfection group and blank control group (t=4.785, P<0.05). For the migration experiment, the migrating number of MG63 cells was (154.36±13.85), (196.32±13.52) and (258.63±12.35), respectively (t=3.968, P<0.01), and that of HOS cells was (131.24±10.58), (247.36±17.52) and (224.52±14.52) respectively (t=3.052, P<0.01). For the invasion experiment, the penetrating number of MG63 cells was (73.50±13.89), (98.63±12.57) and (112.30±15.29), respectively (t=23.178, P<0.01), and that of HOS cells was (70.74±13.49) (98.40±16.46) and 89.74±12.47, respectively (t=14.275, P<0.01). The migration and invasion ability of MG63 and HOS cells in lncRNA HIF2PUT group was lower than that in blank transfection group and blank control group. \u0000 \u0000 \u0000Conclusion \u0000The expression of lncRNA HIF2PUT was significantly low in osteosarcoma tissues and MG63 and HOS cell lines. The expression of lncRNA HIF2PUT significantly inhibited the proliferation, migration and invasion of osteosarcoma cells. \u0000 \u0000 \u0000Key words: \u0000Osteosarcoma; Long-chain non-coding RNA; Migration; Invasion","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"169-171"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49187577","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.1001-9030.2020.01.031
X. Yang, Y. Zhang, Yilun Wu, Lin Wang, Zi-run Wang
Objective To observe the neuraminidase 3 (NEU3) expression and location in SW1353 cells and investigate the effect of NEU3 expression on the proliferation and migration of human chondrosarcoma SW1353 cells. Methods The NEU3 overexpression cell line was constructed by the lipo 3000 transfection reagent. The experiment was divided into 4 groups: UN, and no treatment was given. Blank, was added with equal dose transfection reagent.pcon, was added with no-load plasmid. pNEU3, NEU3 gene overexpression plasmid was added. Immunofluorescence was used to determine the expression of NEU3 in SW1353 cells. Different groups of cells were added the lipo3000 transfection reagent and to explore these groups' mRNA expression of NEU3 by Real-time polymerase chain reaction (PCR). The impact of NEU3 over-expression on the migration of SW1353 cells was examined by scratch test. The effect of NEU3 over-expression on the proliferation of SW1353 cells was measured by EdU proliferation test and CCK-8 assay. The difference in the NEU3 expression and cell capability was tested by two independent samples t test. Results After transfection with NEU3 gene over-expressed plasmid, Real-time PCR showed that NEU3 gene mRNA was significantly up-regulated. EdU proliferation test showed that NEU3 gene over-expression could promote proliferation of SW1353 proliferation cells [positive rate for control group: (6.56±2.29)%, (11.81±6.49)%, (8.64±3.90)%; for treatment group: (40.57±14.32)% (t=11.355, P<0.05]. The scratch test showed that NEU3 gene over-expression could enhance the migration of SW1353 cells [Remian Scratch area for control group: (18 343±367), (20 803±124), (26 472±483) dpi; for treatment group: (3 320±193) dpi, t=7.265, P<0.05]. Conclusion Over-expression of NEU3 gene can enhance proliferation and migration of human chondrosarcoma SW1353 cells. Key words: Chondrosarcoma; Sialidase; Cell proliferation; Cell migration
{"title":"Effects of neuraminidase 3 gene over-expression on proliferation and migration of chondrosarcoma cells","authors":"X. Yang, Y. Zhang, Yilun Wu, Lin Wang, Zi-run Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.031","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.031","url":null,"abstract":"Objective \u0000To observe the neuraminidase 3 (NEU3) expression and location in SW1353 cells and investigate the effect of NEU3 expression on the proliferation and migration of human chondrosarcoma SW1353 cells. \u0000 \u0000 \u0000Methods \u0000The NEU3 overexpression cell line was constructed by the lipo 3000 transfection reagent. The experiment was divided into 4 groups: UN, and no treatment was given. Blank, was added with equal dose transfection reagent.pcon, was added with no-load plasmid. pNEU3, NEU3 gene overexpression plasmid was added. Immunofluorescence was used to determine the expression of NEU3 in SW1353 cells. Different groups of cells were added the lipo3000 transfection reagent and to explore these groups' mRNA expression of NEU3 by Real-time polymerase chain reaction (PCR). The impact of NEU3 over-expression on the migration of SW1353 cells was examined by scratch test. The effect of NEU3 over-expression on the proliferation of SW1353 cells was measured by EdU proliferation test and CCK-8 assay. The difference in the NEU3 expression and cell capability was tested by two independent samples t test. \u0000 \u0000 \u0000Results \u0000After transfection with NEU3 gene over-expressed plasmid, Real-time PCR showed that NEU3 gene mRNA was significantly up-regulated. EdU proliferation test showed that NEU3 gene over-expression could promote proliferation of SW1353 proliferation cells [positive rate for control group: (6.56±2.29)%, (11.81±6.49)%, (8.64±3.90)%; for treatment group: (40.57±14.32)% (t=11.355, P<0.05]. The scratch test showed that NEU3 gene over-expression could enhance the migration of SW1353 cells [Remian Scratch area for control group: (18 343±367), (20 803±124), (26 472±483) dpi; for treatment group: (3 320±193) dpi, t=7.265, P<0.05]. \u0000 \u0000 \u0000Conclusion \u0000Over-expression of NEU3 gene can enhance proliferation and migration of human chondrosarcoma SW1353 cells. \u0000 \u0000 \u0000Key words: \u0000Chondrosarcoma; Sialidase; Cell proliferation; Cell migration","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"109-111"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48852790","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 study the possibility of protein acetyltransferase gene 10 (NAA10) as a target for breast cancer treatment. Methods Based on METABRIC database, 1974 cases of breast cancer patients were selected, including 718 cases of Luminal A subtype, 488 cases of Luminal B subtype, 240 cases of human epidermalgrowth factor receptor-2 (Her-2) subtype, 329 cases of basal-like subtype, and 199 cases of normal-like subtype. Using METABRIC database, the relationship between 33 acetyltransferase genes and the prognosis of breast cancer, and the relationship between NAA10 gene expression and clinicopathological parameters of breast cancer were studied. NAA10 small interfering RNA (siRNA) was used to treat breast cancer cell lines with NAA10 high expression, and the effect of NAA10 gene silencing on breast cancer cell proliferation was detected by cell counting kit-8 (CCK-8) and plate clone formation assays. Results The mRNA expression of NAA10 was significantly correlated with the disease-free survival rate (DFS) and overall survival rate of breast cancer (P<0.05), and the relative risk ratios were 1.31 (1.14-1.51) and 1.13 (1.01-1.25), respectively. NAA10 gene expression was positively correlated with breast cancer histological grade, lymph node status, and Nottingham index (NPI) (P<0.01). NAA10 mRNA and protein levels were highest in Basal-like breast cancer. Compared with control group, NAA10 gene silencing inhibited 50% breast cancer cell proliferation (P<0.05). Conclusion NAA10 gene is closely related to breast cancer cell proliferation and its expression level is closely related to breast cancer prognosis. NAA10 gene may become a target for future treatment of breast cancer. Key words: N-α-acetyltransferase gene 10; Lysine acetyltransferase; Small interfering RNA; Breast cancer
{"title":"Expression and prognosis of N-α-acetyltransferase gene 10 in breast cancer","authors":"Xu-hui Guo, Heng-wei Zhang, Juntao Li, P. Tian, Zeng‐Quan Yang, Xudong Wei","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.040","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.040","url":null,"abstract":"Objective \u0000To study the possibility of protein acetyltransferase gene 10 (NAA10) as a target for breast cancer treatment. \u0000 \u0000 \u0000Methods \u0000Based on METABRIC database, 1974 cases of breast cancer patients were selected, including 718 cases of Luminal A subtype, 488 cases of Luminal B subtype, 240 cases of human epidermalgrowth factor receptor-2 (Her-2) subtype, 329 cases of basal-like subtype, and 199 cases of normal-like subtype. Using METABRIC database, the relationship between 33 acetyltransferase genes and the prognosis of breast cancer, and the relationship between NAA10 gene expression and clinicopathological parameters of breast cancer were studied. NAA10 small interfering RNA (siRNA) was used to treat breast cancer cell lines with NAA10 high expression, and the effect of NAA10 gene silencing on breast cancer cell proliferation was detected by cell counting kit-8 (CCK-8) and plate clone formation assays. \u0000 \u0000 \u0000Results \u0000The mRNA expression of NAA10 was significantly correlated with the disease-free survival rate (DFS) and overall survival rate of breast cancer (P<0.05), and the relative risk ratios were 1.31 (1.14-1.51) and 1.13 (1.01-1.25), respectively. NAA10 gene expression was positively correlated with breast cancer histological grade, lymph node status, and Nottingham index (NPI) (P<0.01). NAA10 mRNA and protein levels were highest in Basal-like breast cancer. Compared with control group, NAA10 gene silencing inhibited 50% breast cancer cell proliferation (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000NAA10 gene is closely related to breast cancer cell proliferation and its expression level is closely related to breast cancer prognosis. NAA10 gene may become a target for future treatment of breast cancer. \u0000 \u0000 \u0000Key words: \u0000N-α-acetyltransferase gene 10; Lysine acetyltransferase; Small interfering RNA; Breast cancer","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"137-140"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42227077","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.1001-9030.2020.01.057
Yueming Zhang, Yaolin Xu, W. Lou
Pancreatic cancer is a highly lethal disease due to rapid progression and poor response to chemotherapy and radiotherapy, and prognosis of patients with pancreatic cancer remains grim. Advances in tumor microenvironment researches uncovered the pivotal role of tumor associated macrophages (TAMs) in tumor microenvironment. Macrophages are polarized into TAMs expressing the M2 phenotype in response to the action of various cytokines and signaling pathways in the tumor microenvironment. TAMs could regulate the development of pancreatic cancer through multiple mechanisms, including promoting immune escape, fibrosis, angiogenesis, cell proliferation and cell invasion. In this study, we reviewed the mechanisms by which TAMs promote the progression of pancreatic ductal adenocarcinoma. Key words: Pancreatic ductal adenocarcinoma; Tumor microenvironment; Tumor associated macrophages; Immune escape
{"title":"Mechanism of tumor-associated macrophages promoting the progression of pancreatic cancer","authors":"Yueming Zhang, Yaolin Xu, W. Lou","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.057","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.057","url":null,"abstract":"Pancreatic cancer is a highly lethal disease due to rapid progression and poor response to chemotherapy and radiotherapy, and prognosis of patients with pancreatic cancer remains grim. Advances in tumor microenvironment researches uncovered the pivotal role of tumor associated macrophages (TAMs) in tumor microenvironment. Macrophages are polarized into TAMs expressing the M2 phenotype in response to the action of various cytokines and signaling pathways in the tumor microenvironment. TAMs could regulate the development of pancreatic cancer through multiple mechanisms, including promoting immune escape, fibrosis, angiogenesis, cell proliferation and cell invasion. In this study, we reviewed the mechanisms by which TAMs promote the progression of pancreatic ductal adenocarcinoma. \u0000 \u0000Key words: \u0000Pancreatic ductal adenocarcinoma; Tumor microenvironment; Tumor associated macrophages; Immune escape","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"183-186"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45139612","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.1001-9030.2020.01.048
D. Haitao, F. Cheng, Chang-mao Liu, Yuan-hua Liu, Zhong Zhang, Changwei Peng, Zhongyu Wang, Jiang Zheng, Chenglong Li
Objective To investigate the expression of human cartilage glycoprotein 39 (YKL-40) in renal clear cell carcinoma and peripheral blood, and to analyze its correlation with prognosis. Methods A total of 110 patients who underwent radical nephrectomy were enrolled as the observation group, and 50 patients with normal physical examination results were enrolled as the control group. The renal clear cell carcinoma tissues and adjacent tissues were taken after operation, and the expression level of YKL-40 mRNA was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR). Results The level of YKL-40 [(186.9±15.7) μg/L] in the peripheral blood of the observation group before surgery was significantly higher than that of the control group [(115.1±12.5) μg/L] and after surgery [(146.3±13.5) μg/L, t=20.565, 28.478, P 0.05). Multivariate Cox analysis showed that Fuhrman classification [OR(95%CI): 2.335(1.359-10.656)], lymph node metastasis [OR(95%CI): 1.854(1.289-7.659)] and peripheral blood YKL-40 [OR(95%CI): 1.335(1.185-6.987)] were independent factors of OS (P<0.05). Conclusion The level of YKL-40 in peripheral blood of patients with renal clear cell carcinoma is elevated and closely related to OS, which may be a prognostic marker. There was no increase in the level of YKL-40 in cancer tissues, and serum YKL-40 may not be derived from cancer cells. Key words: YKL-40; Renal clear cell carcinoma; Overall survival
{"title":"Expression of human cartilage glycoprotein 39 in peripheral blood and tissues in renal cell carcinoma","authors":"D. Haitao, F. Cheng, Chang-mao Liu, Yuan-hua Liu, Zhong Zhang, Changwei Peng, Zhongyu Wang, Jiang Zheng, Chenglong Li","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.048","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.048","url":null,"abstract":"Objective \u0000To investigate the expression of human cartilage glycoprotein 39 (YKL-40) in renal clear cell carcinoma and peripheral blood, and to analyze its correlation with prognosis. \u0000 \u0000 \u0000Methods \u0000A total of 110 patients who underwent radical nephrectomy were enrolled as the observation group, and 50 patients with normal physical examination results were enrolled as the control group. The renal clear cell carcinoma tissues and adjacent tissues were taken after operation, and the expression level of YKL-40 mRNA was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR). \u0000 \u0000 \u0000Results \u0000The level of YKL-40 [(186.9±15.7) μg/L] in the peripheral blood of the observation group before surgery was significantly higher than that of the control group [(115.1±12.5) μg/L] and after surgery [(146.3±13.5) μg/L, t=20.565, 28.478, P 0.05). Multivariate Cox analysis showed that Fuhrman classification [OR(95%CI): 2.335(1.359-10.656)], lymph node metastasis [OR(95%CI): 1.854(1.289-7.659)] and peripheral blood YKL-40 [OR(95%CI): 1.335(1.185-6.987)] were independent factors of OS (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The level of YKL-40 in peripheral blood of patients with renal clear cell carcinoma is elevated and closely related to OS, which may be a prognostic marker. There was no increase in the level of YKL-40 in cancer tissues, and serum YKL-40 may not be derived from cancer cells. \u0000 \u0000 \u0000Key words: \u0000YKL-40; Renal clear cell carcinoma; Overall survival","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"166-168"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42320591","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.1001-9030.2020.01.041
Yong Xu, Rongchao Ying
Objective To explore the expression and significance of X chromosome linked inhibitor of apoptosis protein (XIAP), high mobility group box-1 protein (HMGB1) and Vimentin in gastric cancer. Methods The expression levels of XIAP, HMGB1 and Vimentin in 84 cases of gastric cancer and adjacent normal tissues were detected by immunohistochemical in June 2016 to January 2019 in Hangzhou Dajiangdong Hospital, and the relationship between the expression of XIAP, HMGB1 and Vimentin and clinicopathological parameters was analyzed. Results The positive expression rate of XIAP in gastric cancer (65.48%, 55/84) was higher than that in adjacent normal tissues (17.86%, 15/84, χ2=50.837, P<0.05). The positive expression rate of HMGB1 in gastric cancer tissues (75.00%, 63/84) was higher than that in adjacent normal tissues (9.52%, 8/84, , χ2=73.791, P<0.05). The positive expression rate of Vimentin in gastric cancer tissues (70.23%, 59/84) was higher than that in adjacent normal tissues (20.24%, 17/84, χ2=42.384, P<0.05). The positive rate of XIAP, HMGB1 and Vimentin in stage Ⅲ-Ⅳ of gastric cancer was significantly higher than that in stage Ⅰ-Ⅱ (χ2=12.574, 9.821, P<0.05). The positive expression rate of XIAP, HMGB1 and Vimentin in patients with lymph node metastasis was significantly higher than that in patients without lymph node metastasis (χ2=23.290, 25.337, P<0.05). The positive rate of XIAP, HMGB1 and Vimentin in gastric cancer tissues were not correlated with age, sex and degree of differentiation. There was positive correlation between XIAP and HMGB1 (r=0.597, P<0.01), between XIAP and Vimentin (r=0.621, P<0.01), and between HMGB1 and Vimentin (r=0.634, P<0.01). Conclusion XIAP, HMGB1 and Vimentin were highly expressed in gastric cancer tissues, which could be used as predictors of the occurrence and malignancy of gastric cancer. Key words: X chromosome linked inhibitor of apoptosis protein; High mobility group box-1 protein; Vimentin; Gastric cancer
{"title":"Expression and significance of X chromosome linked inhibitor of apoptosis protein, high mobility group box-1 protein and Vimentin in gastric cancer","authors":"Yong Xu, Rongchao Ying","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.041","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.041","url":null,"abstract":"Objective \u0000To explore the expression and significance of X chromosome linked inhibitor of apoptosis protein (XIAP), high mobility group box-1 protein (HMGB1) and Vimentin in gastric cancer. \u0000 \u0000 \u0000Methods \u0000The expression levels of XIAP, HMGB1 and Vimentin in 84 cases of gastric cancer and adjacent normal tissues were detected by immunohistochemical in June 2016 to January 2019 in Hangzhou Dajiangdong Hospital, and the relationship between the expression of XIAP, HMGB1 and Vimentin and clinicopathological parameters was analyzed. \u0000 \u0000 \u0000Results \u0000The positive expression rate of XIAP in gastric cancer (65.48%, 55/84) was higher than that in adjacent normal tissues (17.86%, 15/84, χ2=50.837, P<0.05). The positive expression rate of HMGB1 in gastric cancer tissues (75.00%, 63/84) was higher than that in adjacent normal tissues (9.52%, 8/84, , χ2=73.791, P<0.05). The positive expression rate of Vimentin in gastric cancer tissues (70.23%, 59/84) was higher than that in adjacent normal tissues (20.24%, 17/84, χ2=42.384, P<0.05). The positive rate of XIAP, HMGB1 and Vimentin in stage Ⅲ-Ⅳ of gastric cancer was significantly higher than that in stage Ⅰ-Ⅱ (χ2=12.574, 9.821, P<0.05). The positive expression rate of XIAP, HMGB1 and Vimentin in patients with lymph node metastasis was significantly higher than that in patients without lymph node metastasis (χ2=23.290, 25.337, P<0.05). The positive rate of XIAP, HMGB1 and Vimentin in gastric cancer tissues were not correlated with age, sex and degree of differentiation. There was positive correlation between XIAP and HMGB1 (r=0.597, P<0.01), between XIAP and Vimentin (r=0.621, P<0.01), and between HMGB1 and Vimentin (r=0.634, P<0.01). \u0000 \u0000 \u0000Conclusion \u0000XIAP, HMGB1 and Vimentin were highly expressed in gastric cancer tissues, which could be used as predictors of the occurrence and malignancy of gastric cancer. \u0000 \u0000 \u0000Key words: \u0000X chromosome linked inhibitor of apoptosis protein; High mobility group box-1 protein; Vimentin; Gastric cancer","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"141-143"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43365137","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.1001-9030.2020.01.058
Pengbo Lyu, Xu Kong, Xiaojie Zhang, Wen-gang Li
For many years, the research on human pancreatic cancer has been based on the traditional two-dimensional metabolism model or genetic engineering mouse model. But these models can not accurately reflect the characteristics of pancreatic cancer and the status of pancreatic cancer microenvironment. As a novel three-dimensional culture technique in vitro, organoid has established reliable in vitro tumor models for pancreatic cancer, liver cancer, gastrointestinal cancer, colon adenocarcinoma, Barrett esophagus and many other diseases. It opens up a new horizon for studying the biological characteristics of pancreatic cancer, exploring more specific anti-pancreatic drugs and formulating individualized treatment schemes for pancreatic cancer. In this paper, the application of organoid technology in pancreatic cancer related research is analyzed and prospected. Key words: Organoid; Pancreatic cancer; Screening of anti-tumor drugs; Individualized treatment of pancreatic cancer
{"title":"Application and perspectives of organoids in pancreatic cancer research","authors":"Pengbo Lyu, Xu Kong, Xiaojie Zhang, Wen-gang Li","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.058","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.058","url":null,"abstract":"For many years, the research on human pancreatic cancer has been based on the traditional two-dimensional metabolism model or genetic engineering mouse model. But these models can not accurately reflect the characteristics of pancreatic cancer and the status of pancreatic cancer microenvironment. As a novel three-dimensional culture technique in vitro, organoid has established reliable in vitro tumor models for pancreatic cancer, liver cancer, gastrointestinal cancer, colon adenocarcinoma, Barrett esophagus and many other diseases. It opens up a new horizon for studying the biological characteristics of pancreatic cancer, exploring more specific anti-pancreatic drugs and formulating individualized treatment schemes for pancreatic cancer. In this paper, the application of organoid technology in pancreatic cancer related research is analyzed and prospected. \u0000 \u0000Key words: \u0000Organoid; Pancreatic cancer; Screening of anti-tumor drugs; Individualized treatment of pancreatic cancer","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"186-188"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46943192","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 elucidate the role and mechanism of tumor suppressor microRN (miRNA, miR)-149-5p in metastasis potentials of renal cell carcinoma. Methods GRC-1 cells were divided into miR-149-5p group(transfected with miR-149-5p mimics), miR-NC group(transfected with mimics control), miR-149-5p+ SRPK1 group(co transfected with miR-149-5p mimics and pcDNA3.1-SRPK1), miR-149-5p+ vector group(co transfected with miR-149-5p mimics and pcDNA3.1). The transfection efficiency of miR-149-5p was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The cell invasion and migration ability were measured by Transwell chamber. The levels of N-cadherin and E-cadherin proteins were detected by Western blotting. SRPK1 may be a target gene of miR-149-5p by using online target gene prediction software. Targeting relationship was identified by double luciferase reporting system. Results The levels of miR-149-5p (2.69±0.27 vs. 1.01±0.08) and E-cadherin (0.81±0.09 vs. 0.42±0.05)in the miR-149-5p group were significantly higher than those in the miR-NC group, and invasion number [(70.81±6.35) vs. (110.64±9.67)], migration number [(108.46±9.27)vs. (162.76±12.71)], N-cadherin (0.46±0.05 vs. 0.79±0.11)and SRPK1 (0.45±0.06 vs. 0.92±0.08)proteins levels were significantly lower than miR-NC group, and the difference was statistically significant (FmiR-149-5p=95.385, Finvasion number=20.216, Fmigration number=22.010, FN-cadherin=15.100, FE-cadherin=31.331, FSRPK1=31.785, P 0.05). The level of E-cadherin (0.45±0.04 vs. 0.80±0.08) in the miR-149-5p+ SRPK1 group was significantly lower than that in the miR-149-5p+ vector group, and the invasive number [(99.51±7.48) vs. (71.84±5.37)], migration number [(145.06±11.14) vs. (107.63±10.20)], N-cadherin (0.86±0.10 vs. 0.47±0.04) and SRPK1 (0.89±0.06 vs. 0.50±0.07) proteins levels were significantly higher than the miR-149-5p+ vector group, and the difference was statistically significant (tSRPK1=7.327, tinvasion number=5.205, tmigration number=4.292, tN-cadherin=6.272, tE-cadherin=6.778, P<0.05). Conclusion Tumor suppressor miR-149-5p targeting negative regulation of SRPK1 expression reduces the migration and invasion of renal cancer cells. Key words: MicroRNA-149-5p; Serine-arginine protein kinase 1; Renal cancer; Metastasis; Invasion
{"title":"Effects of microRNA-149-5p modulating serine-arginine protein kinase 1 on invasion and migration of renal cancer cells","authors":"Jianpeng Bi, Zhaohui Gu, Ziyu Feng, Zhankui Jia, Jinjian Yang, Yan-fang Yang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.027","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.027","url":null,"abstract":"Objective \u0000To elucidate the role and mechanism of tumor suppressor microRN (miRNA, miR)-149-5p in metastasis potentials of renal cell carcinoma. \u0000 \u0000 \u0000Methods \u0000GRC-1 cells were divided into miR-149-5p group(transfected with miR-149-5p mimics), miR-NC group(transfected with mimics control), miR-149-5p+ SRPK1 group(co transfected with miR-149-5p mimics and pcDNA3.1-SRPK1), miR-149-5p+ vector group(co transfected with miR-149-5p mimics and pcDNA3.1). The transfection efficiency of miR-149-5p was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The cell invasion and migration ability were measured by Transwell chamber. The levels of N-cadherin and E-cadherin proteins were detected by Western blotting. SRPK1 may be a target gene of miR-149-5p by using online target gene prediction software. Targeting relationship was identified by double luciferase reporting system. \u0000 \u0000 \u0000Results \u0000The levels of miR-149-5p (2.69±0.27 vs. 1.01±0.08) and E-cadherin (0.81±0.09 vs. 0.42±0.05)in the miR-149-5p group were significantly higher than those in the miR-NC group, and invasion number [(70.81±6.35) vs. (110.64±9.67)], migration number [(108.46±9.27)vs. (162.76±12.71)], N-cadherin (0.46±0.05 vs. 0.79±0.11)and SRPK1 (0.45±0.06 vs. 0.92±0.08)proteins levels were significantly lower than miR-NC group, and the difference was statistically significant (FmiR-149-5p=95.385, Finvasion number=20.216, Fmigration number=22.010, FN-cadherin=15.100, FE-cadherin=31.331, FSRPK1=31.785, P 0.05). The level of E-cadherin (0.45±0.04 vs. 0.80±0.08) in the miR-149-5p+ SRPK1 group was significantly lower than that in the miR-149-5p+ vector group, and the invasive number [(99.51±7.48) vs. (71.84±5.37)], migration number [(145.06±11.14) vs. (107.63±10.20)], N-cadherin (0.86±0.10 vs. 0.47±0.04) and SRPK1 (0.89±0.06 vs. 0.50±0.07) proteins levels were significantly higher than the miR-149-5p+ vector group, and the difference was statistically significant (tSRPK1=7.327, tinvasion number=5.205, tmigration number=4.292, tN-cadherin=6.272, tE-cadherin=6.778, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000Tumor suppressor miR-149-5p targeting negative regulation of SRPK1 expression reduces the migration and invasion of renal cancer cells. \u0000 \u0000 \u0000Key words: \u0000MicroRNA-149-5p; Serine-arginine protein kinase 1; Renal cancer; Metastasis; Invasion","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"93-96"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48538209","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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