The influence of silencing miRNA-155 on proliferation and apoptosis of human cutaneous squamous cell carcinoma cell line A431 / 天津医药

ABSTRACT

Objective To investigate the effects of antisense oligonucleotide against miRNA-155 (AS-miRNA-155) on proliferation,apoptosis and invasion and migration abilities of human cutaneous squamous cell carcinoma cell line A431. Methods AS-miRNA-155 was transfected into human cutaneous squamous cell carcinoma A431 cells by using LipofectamineTM 2000. Blank control without transfection and transfected with non-sense sequence were used as non-sense sequence control. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of miRNA-155 in A431 cells. Cell proliferation was analyzed using dimethyl thiazolyldiphenyl tetrazolium (MTT) assay. Cell cycle arrest and apoptosis were studied by flow cytometry (FCM). Invasion and migration were measured by Transwell chamber assays. Results The relative expression of miRNA-155 mRNA was lower in the transfection group than that in the blank control group and the negative control group (F=634.57, P<0.01), but there was no significant difference between the blank control group and the negative control group. After 72 h transfection, the survival rate was significantly lower in the transfection group than that of the blank control group and the negative control group, and the transfection rate decreased significantly by 120 h (P<0.05). Cells of G0/G1 phase increased, Cells of S phase reduced, the overall PI value decreased in transfection group, and the apoptosis rate of A431 cells, migration and invasion of cells increased (P<0.05). There was no significant difference in G2/M cycle between transfection group, blank control group and negative control group. There were no significant differences in A431 cell apoptosis rate, cell migration and invasive ability between blank control group and negative control group. Conclusion Antisense oligonucleotide against miRNA-155 can inhibit the expression of miRNA-155, the proliferation and promote the apoptosis of human cutaneous squamous cell carcinoma A431 cells, which indicates that miRNA-155 may become a new target for the regulation of gene expression in cutaneous squamous cell carcinoma.