miR-1273g-3p affects proliferation,migration and invasion of colorectal cancer cells by regulating TGF-β/Smad4 signaling pathway
Objective To investigate the effect of miR-1273g-3p on the biological function of colorectal cancer cells by regulating transforming growth factor-β(TGF-β)/mothers against decapentaplegic homolog 4(Smad4)signaling pathway.Methods The expression of miR-1273g-3p in the colorectal cancer HCT116 and SW480 cells was detected by qRT-PCR,and the expression of TGF-β and Smad4 proteins in the two cell lines was detected by Western blot.Human colon epithelial cells NCM460 were used as the control.miR-1273g-3p inhibitor was transfected into HCT116 and SW480 cells(miR-1273g-3p inhibitor group),and an empty control group(inhibitor control group)was set up.qRT-PCR was used to verify the transfection effect.The transfected HCT116 cells were subcutaneously injected into BALB/c nude mice.qRT-PCR and Western blot were used to detect the mRNA and protein expressions of TGF-β and Smad4,respectively,in each group of cells after transfection.The proliferation,migration and invasion of cells were detected by Cell Counting Kit-8(CCK-8),scratch assay and transwell chamber invasion assay.The correlation between miR-1273g-3p and TGF-β was predicted by BiBiServ and TargetScan website,and the binding of miR-1273g-3p to TGF-β was verified by dual-luciferase reporter gene assay.Results Compared with human colon epithelial cells NCM460,miR-1273g-3p,TGF-β protein and Smad4 protein were all highly expressed in human col-orectal cancer cells HCT1 16 and SW480(all P＜0.01).After transfection for 48 hours,compared with the inhibitor control group,the ex-pression of miR-1273g-3p in the miR-1273g-3p inhibitor group was significantly decreased(P＜0.01),the mRNA and protein expressions of TGF-β and Smad4 were significantly decreased(all P＜0.01),and the cellular proliferation,migration and invasion were significantly reduced(all P＜0.01).The results of the dual-luciferase reporter gene assay showed that miR-1273g-3p could target and bind TGF-β,and miR-1273g-3p inhibited the luciferase activity in SW480 cells transfected with wild-type TGF-β-3'untranslated region(UTR)plasmids(P＜O.01),but had no effect on the luciferase activity in SW480 cells transfected with mutant TGF-β-3'UTR plasmids(P＞0.05).Conclusions Inhibiting the expression of miR-1273g-3p can significantly inhibit the proliferation,migration and invasion of colorectal cancer cells HCT116 and SW480,which may be related to the TGF-β/Smad4 signaling pathway.
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