Effects Ofthe Knock-down Of KCTD10 On The Characteristics Ofliver Cancer Stem Cell Like Cells

Objective:This study aimsto explore the effects of the expression of KCTD10 on the characteristics ofliver cancer stem cell like cells and the molecular mechanism involved.Methods:Human hepatoma cell line Hep3B cells and MHCC97H cells were infected by lentivirus containing KCTD 10 siRNA sequence.The inhibitory effect of KCTD 10 expression was confirmed by qPCR and Western blotting(WB)analysis.The sphere formation rate determination experiment,plate colony formation assay,cell proliferation assay and wound healing assay were used to analyze in vitro tumorigenicity,self-renewal ability and the ability of cell migration in hepatocellular carcinoma cellsafter inhibition of KCTD10 expression,respectively.qPCR,WB and fluorescent antibody labeling and flow cytometry were performed to examine the expressions of cancer stem cell markers and the molecule involved in Notch signaling pathway.Results:The transfection by lentivirus expressing KCTD 10 siRNA can effectively inhibit KCTD 10 mRNA and protein levels in Hep3B and MHCC97H cells.Inhibition of the expression of KCTD 10 can increase the tumor sphere formation rate,plate colony formation ability,cell proliferation rate and cell migration ability,and the mRNA levels of liver cancer stem cell marker genes CD133,OCT4 and Bmil,and liver cancer stem cell marker gene to transcription factor Nanog,aldehyde dehydrogenase 1 family member A1(ALDH1A1)protein level,as well as enhance the proportion of CD133+cells in hepatocellular carcinoma cells.At the same time,inhibition of the expression of KCTD10 protein affects Notch signaling pathway regulation in hepatocellular cells.Conclusion:1.Inhibiting the expression of Kctd10 gene increases the tumorigenicity of liver cancer cells.2.Inhibition of Kctd10 gene expression can enhance the characteristics of liver cancer stem cells.3.Inhibition of Kctd10 gene expression affects Notch signaling pathway regulation in hepatocellular cells.