miRNA-30a-5p过表达对肾癌细胞株786-0增殖、凋亡的影响

发布时间：2018-06-10 14:29

本文选题：肾细胞癌 + 微小RNA-a-p　；参考：《山东医药》2017年38期

【摘要】：目的观察miRNA-30a-5p过表达对肾癌细胞株786-0增殖、凋亡的影响,并探讨其机制。方法将肾癌细胞株786-0分为模拟物组、抑制物组、未转染组,模拟物组转染miRNA-30a-5p模拟物,抑制物组转染miRNA-30a-5p抑制物,未转染组为正常肾癌细胞株786-0。采用实时荧光定量PCR法检测模拟物组、抑制物组、未转染组肾癌细胞株786-0及正常人肾小管上皮细胞HK-2(正常组)miRNA-30a-5p,CCK-8法观察各组培养24、48、72 h后的细胞增殖能力,流式细胞术观察前三组细胞凋亡情况,Western blotting法检测前三组细胞中异黏蛋白(MTDH)表达。结果正常组、模拟物组、抑制物组、未转染组细胞miRNA-30a-5p mRNA相对表达量分别为1.00±0.00、1.32±0.05、0.32±0.05、0.52±0.06,正常组、模拟物组、抑制物组与未转染组相比,P均0.01。正常组在培养24、48、72 h后的吸光度值分别为0.26±0.02、0.44±0.03、0.73±0.06,模拟物组分别为0.33±0.04、0.50±0.05、0.88±0.05,抑制物组分别为0.86±0.01、1.88±0.12、2.48±0.15,未转染组分别为0.62±0.01、1.63±0.02、2.34±0.23,正常组、模拟物组各时点分别与抑制物组、未转染组相比,P均0.01。模拟物组、抑制物组、未转染组细胞凋亡率分别为11.91%±0.45%、5.23%±0.38%、6.77%±0.52%,模拟物组与抑制物组、未转染组相比,P均0.01。模拟物组、抑制物组、未转染组MTDH蛋白的相对表达量分别为1.14±0.08、6.31±0.24、5.56±0.06,模拟物组与抑制物组、未转染组相比,P均0.01。结论 miRNA-30a-5p过表达可明显抑制肾癌细胞株786-0的增殖、促进细胞凋亡,其可能是通过调控MTDH蛋白表达实现的。
[Abstract]:Objective to investigate the effect of miRNA-30a-5p overexpression on proliferation and apoptosis of renal carcinoma cell line 786-0 and its mechanism. Methods Renal carcinoma cell line 786-0 was divided into three groups: mimic group, inhibitor group, non-transfection group, mimic group, miRNA-30a-5p mimic group, the inhibitor group transfected miRNA-30a-5p inhibitor, and the untransfected group was normal renal carcinoma cell line 786-0. The proliferation of renal carcinoma cell line 786-0 and normal human renal tubular epithelial cells HK-2 (normal group) was detected by real-time fluorescent quantitative PCR. The proliferation of the cells in each group was observed after cultured for 244872 hours. The apoptosis of the first three groups was observed by flow cytometry and the expression of isoproterenol (MTDH) in the former three groups was detected by Western blotting assay. Results the relative expression levels of miRNA-30a-5p mRNA in normal group, mimic group, inhibitor group and untransfected group were 1.00 卤0.005 + 1.32 卤0.05N 0.32 卤0.05N 0.52 卤0.06, respectively. The expression of miRNA-30a-5p mRNA in normal group, mimic group, inhibitor group and untransfected group were 0.01, respectively. The absorbance values of the control group were 0.26 卤0.02 卤0.03 卤0.73 卤0.06, 0.33 卤0.04 卤0.50 卤0.050.88 卤0.05, 0.86 卤0.01, 1.88 卤0.122.48 卤0.15 and 0.62 卤0.011 卤1.63 卤0.022.34 卤0.23 of the normal group, 0.33 卤0.04 卤0.04 卤0.50 卤0.055 卤0.05, 0.86 卤0.01 卤1.88 卤0.122.48 卤0.15, 0.62 卤0.011 卤1.63 卤0.022.34 卤0.23, respectively. The apoptotic rates of the mimic group, the inhibitor group and the untransfected group were 11.91% 卤0.45 and 5.23% 卤0.38 卤6.77% 卤0.52 respectively. The relative expression of MTDH protein in the mimic group, the inhibitor group and the untransfected group were 1.14 卤0.08t6.31 卤0.2455.56 卤0.06, respectively. Compared with the control group and the untransfected group, the expression of MTDH protein in the mimic group was significantly higher than that in the control group (P < 0.01). Conclusion the overexpression of miRNA-30a-5p can significantly inhibit the proliferation of RCC cell line 786-0 and promote cell apoptosis, which may be achieved by regulating the expression of MTDH protein.
【作者单位】：山东医学高等专科学校;山东医学高等专科学校附属医院;
【基金】：山东省高等学校科技计划项目(J13LL08) 山东省临沂市科技发展计划项目(201515055)
【分类号】：R737.11

【相似文献】