miR-153-3p regulates malignant biological behaviors of esophageal cancer cells by targeting ACTN4
The study aims to explore the effects of miR-153-3p on the proliferation,migration and invasion of esophageal cancer(ESCA)cells by targeting and regulating a-actinin 4(ACTN4).The expression of miR-153-3p and ACTN4 mRNA in different cell lines was detected by qRT-PCR.The targeting relationship between miR-153-3p and ACTN4 was verified by double luciferase assay.ECA109 cells were cultured in vitro and divided into six groups:blank group,mimic control(miR-NC)group,mimic(miR-153-3p)group,empty vector group,ACTN4 overexpression(ACTN4)group,the mimic and ACTN4 overexpression(miR-153-3p+ACTN4)group.qRT-PCR and Western blotting were performed to measure the expression of miR-153-3p and ACTN4;CCK-8 and colony formation experiment was performed to measure the proliferation of cells;Transwell was performed to measure the migration and invasion of cells.Western blotting was used to detect the protein levels of Ki67,E-cadherin,and N-cadherin.A nude mouse model of subcutaneous xenograft tumors was constructed in vivo and the effect of up-regulation of miR-153-3p on tumor growth was detected.Immunohistochemistry was performed to measure the expression levels of ACTN4,Ki67,E-cadherin and N-cadherin in tumor tissues.The results showed that,the expression of miR-153-3p in the ESCA cell line was low(P＜0.05),and ACTN4 mRNA was highly expressed(P＜0.05).miR-153-3p was able to negatively regulate ACTN4 expression(P＜0.05).Overexpression of miR-153-3p can reduce cell viability,colony formation,migration and invasion,inhibit Ki67 and N-cadherin expression,and up regulate E-cadherin expression(P＜0.05).ACTN4 overexpression showed the opposite effects(P＜0.05);overexpression of ACTN4 can reverse the effects of miR-153-3p overexpression on ESCA cell behavior(P＜0.05).In vivo studies have shown that overexpression of miR-153-3p can inhibit the expression of ACTN4,Ki67,N-cadherin,increase the expression of E-cadherin,and slow down tumor growth.This study suggests that overexpression of miR-153-3p may inhibit the proliferation,invasion and migration of ESCA cells by targeting ACTN4 negatively.
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