摘要
目的:探讨柴胡皂苷D调控miR-34a对胶质瘤细胞C6增殖与自噬的影响。方法:分别以终浓度为0、3、6、9、12、15μmol/L的柴胡皂苷D作用胶质瘤细胞C6后,采用CCK-8法检测细胞增殖能力;Western blot和RT-qPCR检测胶质瘤细胞C6中自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值及Beclin1表达情况。将胶质瘤细胞C6分成4组:对照组(Control组)、柴胡皂苷D组(Ssd组)、柴胡皂苷D+miR-34a阴性对照转染组(Ssd+miR-34a NC组)、柴胡皂苷D+miR-34a inhibitor转染组(Ssd+miR-34a inhibitor组),CCK-8法检测miR-34a NC和miR-34a inhibitor在柴胡皂苷D处理后对胶质瘤细胞C6增殖能力的影响;Western blot检测各组胶质瘤细胞C6中自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ和Beclin-1的蛋白表达情况。结果:经过不同浓度的柴胡皂苷D处理48 h后,胶质瘤细胞C6增殖能力明显下降,自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值和Beclin1表达水平均升高(P<0.05);胶质瘤细胞C6中miR-34a表达水平升高(P<0.05)。转染后,Ssd组胶质瘤细胞C6增殖率、LC3-Ⅱ/LC3-Ⅰ比值和Beclin1表达水平与Ssd+miR-34a NC组比较,差异无统计学意义(P>0.05);Ssd组、Ssd+miR-34a NC组胶质瘤细胞C6增殖率低于Control组和Ssd+miR-34a inhibitor组,LC3-Ⅱ/LC3-Ⅰ比值和Beclin1表达水平均高于Control组和Ssd+miR-34a inhibitor组,差异均有统计学意义(P<0.05);Ssd+miR-34a inhibitor组胶质瘤细胞C6增殖率低于Control组,LC3-Ⅱ/LC3-Ⅰ比值高于Control组,差异均有统计学意义(P<0.05)。结论:柴胡皂苷D可能是通过调控miR-34a来抑制胶质瘤细胞C6增殖和诱导自噬发生。
Objective:To investigate the effect of saikosaponin D regulating miR-34a on the proliferation and autophagy of glioma cell line C6.Methods:Take the final concentration as 0,3,6,9,12 and 15μmol/L after treating glioma cell C6 with saikosaponin D,the cell proliferation ability was detected by CCK-8,and the autophagy related proteins Beclin1,LC3-II/LC3-I,and gene expression in glioma cell C6 were detected by Western blot and qRT PCR.Divide glioma cell C6 into four groups:control group(Control group),saikosaponin D group(Ssd group),saikosaponin D+miR-34a negative control transfection group(Ssd+miR-34a NC group),saikosaponin D+miR-34a inhibitor transfection group(Ssd+miR-34a inhibitor group),and detected the effects of miR-34a NC and miR-34a inhibitor on the proliferation ability of glioma cell C6 after treatment with saikosaponin D,Western blot was used to detect the protein expression of autophagy related proteins LC3-Ⅰ,LC3-Ⅱ,and Beclin-1 in C6 glioma cells of each group.Results:After treatment with different concentrations of saikosaponin D for 48 h,the proliferation ability of glioma cells C6 was decreased,the ratio of autophagy-related protein LC3II/LC3-I was increased,and the expression level of Beclin1 was increased.There was no statistically significant difference in C6 proliferation rate,LC3 II/LC3-I ratio,and Beclin1 expression level between the Ssd group and the Ssd+miR-34a NC group(P>0.05).The proliferation rate of C6 glioma cells in the Ssd group and Ssd+miR-34a NC group was lower than that in the Control group and Ssd+miR-34a inhibitor group.The LC3 II/LC3-I ratio and Beclin1 expression level were higher than those in the Control group and Ssd+miR-34a inhibitor group,and the differences were statistically significant(P<0.05).The proliferation rate of C6 glioma cells in the Ssd+miR-34a inhibitor group was lower than that in the Control group,and the LC3 II/LC3-I ratio was higher than those in the Control group,with statistical significance(P<0.05).Conclusion:Chaihu saponin D may inhibit the proliferation of glioma cell C6 and induce autophagy by regulating miR-34a.
作者
刘晓熙
赵鹏伟
张静
于慧玲
LIU Xiao-xi;ZHAO Peng-wei;ZHANG Jing;YU Hui-ling(Department of Basic Medicine,Inner Mongolia Medical University,Hohhot 010020,Inner Mongolia,China)
出处
《川北医学院学报》
2024年第1期8-11,17,共5页
Journal of North Sichuan Medical College
基金
内蒙古自治区自然科学基金项目(2019MS08153)。
