MST1 and MST2 in Hippo signaling pathway were critical in stemness maintenance and self-renewal of hepatocellular carcinoma stem cells

Abstract

Abstract: Objective To investigate the role of MST1 and MST2 expressed in Hippo signaling pathway in stemness maintenance and self-renewal of liver cancer stem cells.Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were conducted to detect the expression levels of MST1 and MST2 in hepatocellular carcinoma (HCC) tissue samples. qRT-PCR and western blot were used to detect the expression levels of MST1 and MST2 in HCC cell lines. CD90+, CD105+ and CD133+ SMMC-7721 cells were selected by flow cytometry. Stem cell marker levels were measured by qRT-PCR and western blot. The expression of MST1 and MST2 were knocked down by shRNA. The formation of mammospheres was observed by microscopy.Results MST1 and MST2 were significantly highly expressed in HCC tissue samples and HCC cells. CD90+, the mammosphere-forming ability in CD105+ and CD133+ SMMC-7721 cells were significantly enhanced. The phosphorylation levels of AMPK increased remarkably, and the expression levels of OCT-4 and Nanog were upregulated to 2.56 ± 0.21, 2.32 ± 0.11. Knocking down MST1 or MST2 could reduce the mammosphere-forming ability of CD90+, CD105+, and CD133+ SMMC-7721 cells. And the phosphorylation levels of AMPK were attenuated. The expression levels of OCT-4 and Nanog were down-regulated to 0.48 ± 0.0 and 0.26 ± 0.08 after knocking down the MST1. The expression levels of OCT-4 and Nanog were down-regulated to 0.35 ± 0.06 and 0.42 ± 0.03 after knocking down the MST2.Conclusion There is an important biological significance of MST1 and MST2 in the stemness maintenance and self-renewal of HCC stem cells.

Key words: MST1, MST2, Hepatocellular carcinoma, Stemness maintenance, Self-renewal

LV Shao-dong, WEI Yong-peng, WANG Zhuo, YUAN Jian-yong, LU Jun-hua. MST1 and MST2 in Hippo signaling pathway were critical in stemness maintenance and self-renewal of hepatocellular carcinoma stem cells[J]. Chinese Hepatolgy, 2021, 26(8): 866-870.

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