Effect of BMP5 on biological behaviors of liver cancer cells and its mechanism
Objective To investigate the effect and mechanism of bone morphogenetic protein 5(BMP5)on the bio-logical behaviors of human liver cancer cells.Methods Normal human liver cell line L02 and human liver cancer cell lines Hep3B and Huh7 were cultured in the incubator at 37℃with 5%CO2.The expression of BMP5 mRNA in these cells was detected by real time-quantitive PCR(RT-qPCR).Human liver cancer cells Hep3B and Huh7 were divided into the control group(NC group),low-dose group,medium-dose group and high-dose group.Cells in the low-,medium-and high-dose groups were treated with recombinant human bone morphogenetic protein 5(rh-BMP5)at concentrations of 1,10 and 100 ng/mL,respectively.Cell proliferation was detected by CCK-8 assay and apoptosis was detected by Annexin-V/PI flow cytometry.Scratch assay was used to detect cell migration.Western blotting was performed to detect the expres-sion of proliferating cell nuclear antigen(PCNA)and key molecules in SMAD pathway(SMAD3)and its phosphorylation(p-SMAD3).Results The expression levels of BMP5 in Hep3B and Huh7 cells significantly increased in comparison with that in L02 cells(both P<0.05).Compared with NC group,rh-BMP5 promoted the proliferation of Hep3B and Huh7 cells and up-regulated the expression of PCNA in the medium-dose and high-dose groups(all P<0.05).After the high-dose(100 ng/mL)rh-BMP5 treatment,Hep3B and Huh7 cells showed decreased apoptosis rate and increased migration rate at 24 h and 48 h in comparison with those of the NC group(both P<0.05).The high-dose rh-BMP5 group had higher p-SMAD3 level than the NC group(P<0.05).Conclusions The expression of BMP5 increases in liver cancer cell lines.BMP5 promotes proliferation and migration and inhibits apoptosis of Hep3B and Huh7 cells,which may be mediated by SMAD3 phosphorylation,and thus facilitates tumor progression.
liver carcinomabone morphogenetic protein 5cell proliferationSMAD3
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