The effect of miRNA-34a antisense oligonucleotide on non-small cell lung cancer cell line HCC827

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 miRNA-34a 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 0.05). At 48 h, 72 h and 96 h, the proliferation level of HCC827 cells in anti-sense 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 non-small 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. Key words: Oligonucleotides, antisense; miRNA-34a; Carcinoma, non-small-cell lung; Cell line, tumor