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目的 探讨miRNA-34a反义寡核苷酸对非小细胞肺癌细胞的影响及分子机制.方法 qRT-PCR检测人非小细胞肺癌细胞株HCC827和人正常肺细胞MRC-5中miRNA-34a表达水平.将HCC827细胞分为3组:空白对照组、阴性对照miRNA组(miRNA-NC)、反义寡核苷酸miRNA-34a组(脂质体2000转染反义寡核苷酸miRNA-34a);CCK-8法检测细胞增殖能力,吉姆萨染色检测细胞克隆能力,Transwell实验检测细胞迁移和侵袭能力;RT-PCR和Westem blot检测细胞中磷酸酶和紧张素同系物(PTEN)、磷酸化蛋白激酶B(p-Akt)、磷脂酰肌醇3-激酶(PI3K) mRNA和蛋白表达水平 HCC827细胞miRNA-34a相对表达量明显高于人正常肺细胞,差异有统计学意义(P＜0.01).反义寡核苷酸miRNA-34a组miRNA-34a相对表达量明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P＜0.05);阴性对照miRNA组与空白对照组miRNA-34a相对表达量之间差异无统计学意义(P＞0.05).培养48、72、96 h时反义寡核苷酸miRNA-34a组HCC827细胞增殖水平明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P＜0.05).反义寡核苷酸miRNA-34a组细胞克隆形成率明显低于阴性对照miRNA组、空白对照组,差异有统计学意义(P＜0.01).反义寡核苷酸miRNA-34a组HCC827细胞迁移和侵袭个数明显低于阴性对照miRNA组和空白对照组,差异有统计学意义(P＜0.01).反义寡核苷酸miRNA-34a组细胞PTEN mRNA和蛋白相对表达量明显高于阴性对照miRNA组与空白对照组,p-Akt、PI3K mRNA和蛋白相对表达量明显低于阴性对照miRNA组与空白对照组,差异有统计学意义(P＜0.05).结论 人非小细胞肺癌细胞中miRNA-34a表达水平明显高于人正常肺细胞;miRNA-34a反义寡核苷酸能够抑制人非小细胞肺癌细胞的增殖、克隆、迁移和侵袭,其机制可能与负调控PTEN/p-Akt/PI3K信号通路有关.“,”Objective To investigate the effect of antisense oligonucleotides of miRNA-34a on non-small cell lung cancer (NSCLC) and its molecular mechanism.Methods The expression of miRNA34a in human non-small cell lung cancer cell line HCC827 and human normal lung cell MRC-5 was detected by real time fluorescence quantitative polymerase chain reaction (qRT-PCR).HCC827 cells were divided into three groups:blank control group,negative control group,anti-sense oligonucleotide group (liposome 2000 transfected anti-sense oligonucleotide miRNA-34a);cell counting kit-8 (CCK-8) method was used to detect cell proliferation,Jimsa staining was used to detect cell cloning ability,Transwell test was used to detect cell migration and invasion ability;RT-PCR and Western blot were used to detect phosphatase and tensin homolog (PTEN),phosphorylation-protein kinase B (p-Akt),phosphatidylinositol-3-kinase (PI3K)mRNA and protein expression.Results The relative expression of miRNA34a in HCC827 cells was significantly higher than that in human normal lung cells (P ＜ 0.01).The relative expression of miRNA34a in antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group (P ＜ 0.05),and there was no significant difference between negative control group and blank control group (P ＞ 0.05).At 48 h,72 h and 96 h,the proliferation level of HCC827 cells in antisense oligonucleotide miRNA-34a group was significantly lower than that in negative control group and blank control group (P ＜ 0.05).The cell cloning rate of antisense oligonucleotide miRNA-34a group was significantly lower than that of negative control group and blank control group (P ＜ 0.01).The number of migration and invasion of HCC827 cells in antisense oligonucleotide RNA-34a group was significantly lower than that in negative control group and blank control group (P ＜0.01).The relative expression of PTEN mRNA and protein in antisense oligonucleotide miRNA-34a group was significantly higher than that in negative control group and blank control group (P ＜ 0.05);the relative expression of p-Akt,PI3K mRNA and protein in antisense oligonucleotide miRNA-34a group were significantly lower than that in negative control group and blank control group (P ＜ 0.05).Conclusions The expression level of miRNA-34a in human nonsmall cell lung cancer cells is significantly higher than that in human normal lung cells.Antisense oligonucleotides of miRNA-34a can inhibit the proliferation,cloning,migration and invasion of human non-small cell lung cancer cells.The mechanism may be related to the negative regulation of PTEN/p-Akt/PI3K signaling pathway.