Hippo信号通路中MST1和MST2的表达在肝癌干细胞干性维持和自我更新中的意义

肝脏 ›› 2021, Vol. 26 ›› Issue (8): 866-870.

Hippo信号通路中MST1和MST2的表达在肝癌干细胞干性维持和自我更新中的意义

闾少冬, 魏勇鹏, 王卓, 袁建勇, 卢军华

200438 上海东方肝胆外科医院肝外五科

收稿日期:2021-02-08 出版日期:2021-08-31 发布日期:2021-09-29
通讯作者: 卢军华,Email:lujh0810@163.com

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

LV Shao-dong, WEI Yong-peng, WANG Zhuo, YUAN Jian-yong, LU Jun-hua

Department of hepatic surgery, Shanghai Eastern Hepatobiliary Surgery Hospital, Shanghai 200438, China

Received:2021-02-08 Online:2021-08-31 Published:2021-09-29
Contact: LU Jun-hua,Email:lujh0810@163.com

摘要/Abstract

摘要： 目的初步探索Hippo信号通路中MST1和MST2的表达在肝癌干细胞干性维持和自我更新中的作用。方法 qRT-PCR和免疫组化检测肝癌组织样本中MST1和MST2的表达;qRT-PCR和Western blot检测肝癌细胞中MST1和MST2的表达;通过流式细胞术筛选CD90+、CD105+、CD133+型的SMMC-7721细胞;qRT-PCR和Western blot检测干细胞标志物水平;转染shRNA在肝癌干细胞中敲低MST1和MST2的表达,镜检观察细胞微球体形成情况,并通过qRT-PCR和Western blot检测干细胞标志物水平。结果 MST1和MST2在肝癌组织样本和肝癌细胞中显著高表达;CD90+、CD105+、CD133+型的SMMC-7721细胞微球体形成能力显著增强,干细胞标志物AMPK磷酸化的水平增加,OCT-4和Nanog的表达上调为2.56±0.21、2.32±0.11;敲低MST1和MST2都能显著CD90+、CD105+、CD133+型的SMMC-7721细胞的微球体形成能力,干细胞标志物AMPK磷酸化的水平减弱,敲低MST1后,OCT-4和Nanog的表达下调为0.48±0.02和0.26±0.08;敲低MST1后,OCT-4和Nanog的表达下调为0.35±0.06和0.42±0.03。结论 MST1和MST2的表达在肝癌干细胞的干性维持和自我更新中有着重要的生物学意义。

关键词: MST1, MST2, 肝癌, 干性维持, 自我更新

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

闾少冬, 魏勇鹏, 王卓, 袁建勇, 卢军华. Hippo信号通路中MST1和MST2的表达在肝癌干细胞干性维持和自我更新中的意义[J]. 肝脏, 2021, 26(8): 866-870.

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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