The effect of miRNA-34a antisense oligonucleotide on non-small cell lung cancer cell line HCC827 / 中国医师杂志

ABSTRACT

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 groupsblank 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.