Effects and mechanism of BMP9 on stemness,proliferation and invasion of cancer stem cells in hepatocellular carcinoma
Objective To explore the effects and mechanism of bone morphogenetic protein 9(BMP9)on the stem-ness,proliferation and invasion of cancer stem cells(CSCs)in hepatocellular carcinoma(HCC).Methods Normal liv-er cells,liver carcinoma cells,and liver carcinoma tumor stem cells in the logarithmic growth phase were selected.RT-qP-CR was used to detect BMP9 mRNA expression.Western blotting was used to detect BMP9 protein expression.HepG2-CSCs were transfected with lentiviral interference vectors to knock down BMP9 expression,which were then divided into HepG2-CSCs group and HepG2-CSCs-BMP9 knockdown group.After adding MAPK/ERK signal agonists(DIPQUO),cells were divided into three groups,HepG2-CSCs group,HepG2-CSCs knockdown group,and HepG2-CSCs knockdown+ DIPQUO group.RT-qPCR was used to detect the expression levels of stemness-related molecules CD44,SOX2,and OCT4 in HepG2-CSCs.CCK-8 assay was used to detect cell proliferation ability.Transwell assay was used to detect cell in-vasion ability.Western blotting was used to detect the expression of key proteins in the MAPK/ERK pathway.Results Compared with normal liver cells,BMP9 expression was up-regulated in liver carcinoma cells and HepG2-CSCs(P<0.05).The BMP9 mRNA and protein expression levels in HepG2-CSCs were higher than those in HepG2 cells(both P<0.05).After BMP9 knockdown,the mRNA expression levels of stem cell-related markers CD44,SOX2,and OCT4 in HepG2 CSCs in the HepG2-CSCs-BMP9 knockdown group were lower than those in the HepG2-CSCs group(all P<0.05).Compared with the HepG2-CSCs group,BMP9 knockdown inhibited the cell proliferation ability of HepG2-CSCs in the HepG2-CSCs-BMP9 knockdown group at 48,72,and 96 h(all P<0.05),and inhibited the migration and invasion abili-ties of HepG2-CSCs cells(all P<0.05).Compared with the HepG2-CSCs group,after BMP9 knockdown,the expression levels of p-ERK1/2 and p-MEK1/2 proteins in the HepG2-CSCs-BMP9 knockdown group significantly decreased(all P<0.05).After adding DIPQUO,the expression levels of stem cell-related markers CD44,SOX2,and OCT4 mRNA increased in the HepG2-CSCs knockdown+DIPQUO group(all P<0.05),and the proliferation and invasion abilities of HepG2-CSCs were enhanced(all P<0.05).Conclusion Knockdown of BMP9 can inhibit the dry maintenance and pro-liferation and invasion functions of HepG2 CSCs,and the mechanism may be related to the inhibition of the MAPK/ERK pathway.
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