CAJ | 학술논문

目的探讨miR-137对胶质瘤细胞株U87细胞增殖和凋亡的影响及相关作用机制.方法采用miR-137寡聚核甘酸(miR-137 mimics)转染U87细胞,荧光定量PCR检测转染后细胞中miR-137的表达水平.实验分为空白对照组、阴性对照组和miR-137组.应用噻唑蓝细胞增殖试验(MTT)、流式细胞术和免疫荧光评价miR-137对U87细胞增殖和凋亡的影响,Western blotting法检测miR-137对增殖及凋亡相关蛋白的影响. 结果 MTT分析显示miR-137寡聚核苷酸转染U87细胞48 h后空白对照组、阴性对照组及miR-137组细胞相对存活率分别为(105.16±8.57)％,(98.57±8.21)％和05.84±6.93)％,差异具有统计学意义(F=82.157,P=0.000).磷脂结合蛋白/碘化丙啶(Annexin V/PI)双染色法检测显示细胞凋亡明显升高,达到16.6％,差异有统计学意义(F=63.837,P=0.000).Hoechst33258染色后荧光显微镜观察发现miR-137 mimics组细胞出现凋亡小体.miR-137 mimics显著抑制U87细胞中CDC42、pERK1/2、AKT及pAKT蛋白表达水平. 结论 miR-137可以抑制人脑胶质瘤细胞增殖,并促进其凋亡,有望成为胶质瘤基因治疗的新靶点.
목적 탐토miR-137대효질류세포주U87세포증식화조망적영향급상관작용궤제.방법 채용miR-137과취핵감산(miR-137 mimics)전염U87세포,형광정량PCR검측전염후세포중miR-137적표체수평.실험분위공백대조조、음성대조조화miR-137조.응용새서람세포증식시험(MTT)、류식세포술화면역형광평개miR-137대U87세포증식화조망적영향,Western blotting법검측miR-137대증식급조망상관단백적영향. 결과 MTT분석현시miR-137과취핵감산전염U87세포48 h후공백대조조、음성대조조급miR-137조세포상대존활솔분별위(105.16±8.57)％,(98.57±8.21)％화05.84±6.93)％,차이구유통계학의의(F=82.157,P=0.000).린지결합단백/전화병정(Annexin V/PI)쌍염색법검측현시세포조망명현승고,체도16.6％,차이유통계학의의(F=63.837,P=0.000).Hoechst33258염색후형광현미경관찰발현miR-137 mimics조세포출현조망소체.miR-137 mimics현저억제U87세포중CDC42、pERK1/2、AKT급pAKT단백표체수평. 결론 miR-137가이억제인뇌효질류세포증식,병촉진기조망,유망성위효질류기인치료적신파점.
Objective To explore the effect ofmiR-137 on proliferation and apoptosis of U87 glioma cells.Methods The miR-137 mimics were transfected to human glioma cell line U87 as miR-137 transfection group; blank control group and negative control group were also employed in our study.The expression ofmiR-137 was identified by real-time polymerase chain reaction (PCR) after transfection.Methylthiazol tetrazoliu (MTT) assay,flow cytometry-Annexin V/PI assay and fluorescence microscope were employed to detect the cell proliferation and apoptosis.MiR-137 and its relative targeting protein expressions were detected by Westem blotting.Results MTT assay showed that the relative cell survival rates in the blank control group,negative control group and miR-137 transfection group were (105.16±8.57)％,(98.57±8.21)％ and (45.84±6.93)％,with significant differences (F=82.157,P=0.000).Annexin V/PI assay showed that the cell apoptosis rates in the blank control group,negative control group and miR-137 transfection group were (4.3±0.63)％,(4.7±0.77)％ and (16.6±1.98)％,with significant differences (F=63.837,P=0.000); and apoptotic body was detected by fluorescence microscope.Moreover,CDC42,pERK1/2,AKT and pAKT protein expression levels were inhibited after transfected by miR-137 mimics.Conclusion MiR-137 inhibits proliferation and induces apoptosis of U87 glioma cells,suggesting that miR-137 could be tumor suppressor gene in glioma.