lncRNA PlncRNA-1对胃癌SGC-7901细胞生物学行为及PI3K/AKT信号通路的影响

Effects of lncRNA PlncRNA-1 on biological behavior and PI3K/AKT signaling pathway of gastric cancer SGC-7901 cells

目的探讨长链非编码RNA(lncRNA)PlncRNA-1对胃癌SGC-7901细胞增殖、周期、凋亡和磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路的影响。方法采用实时荧光定量PCR(qPCR)检测PlncRNA-1在SGC-7901细胞和正常胃黏膜上皮GES-1细胞中的表达水平,将体外培养的SGC-7901细胞分为未转染组(正常培养)、阴性组(转染阴性对照siRNA)和干扰组(转染PlncRNA-1-siRNA),qPCR检测SGC-7901细胞中PlncRNA-1表达水平,噻唑蓝(MTT)法检测SGC-7901细胞增殖活力,流式细胞仪检测SGC-7901细胞周期和凋亡情况,免疫印迹法检测PI3K/AKT信号通路相关蛋白PI3K、AKT、磷酸化(p)-AKT、细胞周期素D1(CyclinD1)和B淋巴细胞瘤-2基因(Bcl-2)蛋白表达情况。结果与GES-1细胞比较,SGC-7901细胞中PlncRNA-1表达水平明显升高,差异有统计学意义(P<0.05);与未转染组比较,干扰组细胞中PlncRNA-1表达水平、细胞增殖活力和细胞在S期、G2/M期所占百分比以及PI3K、p-AKT、CyclinD1、Bcl-2蛋白表达水平均明显降低,而细胞凋亡率和细胞在G0/G1期所占百分比明显升高,差异有统计学意义(P<0.05);但阴性组和未转染组之间各指标差异均无统计学意义(P>0.05)。结论PlncRNA-1在胃癌SGC-7901细胞中表达升高,干扰其表达可抑制SGC-7901细胞增殖并诱导细胞周期阻滞和凋亡,其作用机制与抑制PI3K/AKT信号通路的活化有关。

Objective To investigate the effects of long non-coding RNA(lncRNA)PlncRNA-1 on proliferation,cell cycle,apoptosis and phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)signaling pathway in gastric cancer SGC-7901 cells.Methods The expression level of PlncRNA-1 in SGC-7901 cells and normal gastric mucosa GES-1 cells was detected by realtime quantitative PCR(qPCR),and SGC-7901 cells cultured in vitro were divided into three groups:untransfected group(normal culture),negative group(transfected with negative control siRNA)and interference group(transfected with PlncRNA-1-sirna),the expression level of PlncRNA-1 in SGC-7901 cells was detected by qPCR,methyl thiazolyl tetrazolium(MTT)assay was used to detect the proliferation of SGC-7901 cells,cell cycle and apoptosis of SGC-7901 cells were detected by flow cytometry,in addition,the expressions of PI3K,AKT,phosphorylated(P)-AKT,CyclinD1 and Bcl-2 were detected by Western blot.Results Compared with that in GES-1 cells,the expression level of PlncRNA-1 in SGC-7901 cells was significantly higher(P<0.05);compared with those in untransfected group,the PlncRNA-1 expression level,cell proliferation activity and percentages of cells in S phase and G2/M phase,the PI3K,p-AKT,CyclinD1 and Bcl-2 protein expression levels in the interference group were significantly lower,while the apoptosis rate and the percentage of cells in G0/G1 phase were significantly higher(P<0.05);however,there was no significant difference between the negative group and the untransfected group(P>0.05).Conclusions The expression of PlncRNA-1 is increased in gastric cancer SGC-7901 cells.Interference of PlncRNA-1 expression can inhibit the proliferation of SGC-7901 cells and induce cell cycle arrest and apoptosis.The mechanism of PlncRNA-1 is related to the inhibition of PI3K/AKT signaling pathway.