芪玉三龙方干扰miRNA21/PTEN/PI3K信号轴抑制LLC肺癌细胞增殖

Qiyu Sanlong Formula inhibited the proliferation of LLC lung cancer cells by interfering with miRNA21/PTEN/PI3K signal axis

目的:探讨芪玉三龙方抑制LLC肺癌细胞增殖的分子机制。方法:LLC肺癌细胞常规传代培养,分别设置为LLC组、空白血清组、siRNA组、含药血清组。MTT法筛选芪玉三龙方最佳作用浓度和作用时间。RT-qPCR、Western Blot及免疫荧光法检测miRNA21/PTEN/PI3K信号轴相关分子表达。结果:芪玉三龙方含药血清呈浓度依赖性抑制LLC肺癌细胞活性。与LLC组比较,siRNA组和含药血清组miRNA21 mRNA表达显著降低(P<0.01),PTEN蛋白和mRNA表达显著上升(P<0.01),p-AKT、p-mTOR、eIF4E、p70S6K蛋白表达显著降低(P<0.05,P<0.01),含药血清组PI3K蛋白表达显著降低(P<0.05)。结论:芪玉三龙方可抑制miRNA21的转录,使PTEN表达有所上升,下调PI3K/AKT/mTOR信号轴关键分子表达,从而温和抑制LLC肺癌细胞增殖。

Objective:To investigate the molecular mechanism of Qiyu Sanlong Formula in inhibiting the proliferation of LLC lung cancer cells.Methods:LLC lung cancer cells were routinely subcultured and set up as LLC group,blank serum group,siRNA group and drug containing serum group,respectively.MTT method was used to screen the best concentration and time of Qiyu Sanlong Formula.The expression of miRNA21/PTEN/PI3K signal axis related molecules was detected by RT-qPCR,Western Blot and immunofluorescence.Results:Qiyu Sanlong Formula contained serum inhibited the activity of LLC lung cancer cells in a concentration dependent manner.Compared with LLC group,the expression of miRNA21 mRNA in siRNA group and drug containing serum group decreased significantly(P<0.01),the expression of PTEN protein and mRNA increased significantly(P<0.01),the expression of p-AKT,p-mTOR,eIF4E,p70S6K protein decreased significantly(P<0.05,P<0.01),and the PI3K protein in drug containing serum group decreased significantly(P<0.05).Conclusion:Qiyu Sanlong Formula can inhibit the transcription of miRNA21,increase the expression of PTEN,and down regulate the expression of key molecules of PI3K/AKT/mTOR signal axis,thereby mildly inhibiting the proliferation of LLC lung cancer cells.