Page 14 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第43卷第1期
· 8 · Journal of Nanjing Medical University（Natural Sciences） 2023年1月

·基础研究·

YAP通过调控EEF1A2的表达促进肝癌细胞的迁移和侵袭

田 祯，庄皓冉，江楚雯，尹 靖，韦晓莹，张枝林，史 琳，张 育，沈维干 *
扬州大学医学院，江苏 扬州 225000

［摘 要］ 目的：研究 Yes 相关蛋白（yes⁃associated protein，YAP）通过调控真核细胞翻译延伸因子 1A2（eukaryotic translation
elongation factor 1 alpha 2，EEF1A2）的表达促进肝癌细胞迁移和侵袭的机制。方法：通过建立YAP和EEF1A2干扰表达或过表
达的SMMC⁃7721和SK⁃HEP1肝癌细胞模型，采用Transwell实验检测细胞迁移和侵袭能力的改变；应用反转录和实时定量PCR
（reverse transcription⁃quantitative real⁃time polymerase chain reaction，RT⁃qPCR）和Western blot 法检测相关基因的mRNA和蛋白
表达水平；利用染色质靶向酶切检测（cleavage under targets and release using nuclease，CUT&RUN）⁃qPCR实验检测YAP能否与
EEF1A2基因的启动子序列结合。结果：EEF1A2在肝癌组织中高表达，并与肝癌细胞的迁移能力呈正相关；EEF1A2可以促进
低迁移能力的SMMC⁃7721细胞和高迁移能力的SK⁃HEP1细胞迁移和侵袭，调控细胞中上皮间质转化（epithelial⁃mesenchymal
transition，EMT）相关标志物 Snail 和 E⁃cadherin 的表达；YAP 可以上调 EEF1A2 的表达而促进肝癌细胞的迁移和侵袭；
CUT&RUN⁃qPCR实验结果显示YAP可以与EEF1A2基因的启动子序列结合。结论：YAP可以通过与EEF1A2基因的启动子序
列结合，促进EEF1A2的表达，进一步促进肝癌细胞的迁移和侵袭。
［关键词］ 真核细胞翻译延伸因子1A2；Yes相关蛋白；细胞迁移；细胞侵袭；肝癌细胞
［中图分类号］ R735.7 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）01⁃008⁃09
doi：10.7655/NYDXBNS20230102

YAP promotes the migration and invasion of hepatocellular carcinoma cells through
regulating the expression of EEF1A2

TIAN Zhen，ZHUANG Haoran，JIANG Chuwen，YIN Jing，WEI Xiaoying，ZHANG Zhilin，SHI Lin，ZHANG Yu，
SHEN Weigan *
School of Medicine，Yangzhou University，Yangzhou 225000，China

［Abstract］ Objective：To investigate the mechanism of yes⁃associated protein（YAP）in promoting the migration and invasion of
hepatocellular carcinoma cells through regulating the expression of eukaryotic translation elongation factor 1 alpha 2（EEF1A2）.
Methods：YAP and EEF1A2 were knockdown or overexpressed in human hepatocellular carcinoma（HCC）cell lines（SMMC⁃7721 and
SK⁃HEP1）respectively. Transwell assays were performed to examine the changes of the migratory and invasive ability of the HCC
cells. Reverse transcription⁃quantitative real⁃time polymerase chain reaction（RT⁃qPCR）and Western blot were conducted to evaluate
the expression of the indicated genes in HCC cells. Cleavage under targets and release using nuclease（CUT&RUN）⁃qPCR assay was
performed to evaluate whether YAP interacts with the promoter of EEF1A2. Results：Upregulation of EEF1A2 was shown in HCC
tissues，and the expression of YAP and EEF1A2 was positively correlated with the spontaneous migratory potential of HCC cell lines.
EEF1A2 facilitated the migration and invasion of the low migratory potential SMMC7721 cells and the high migratory potential SK⁃HEP1
cells and regulated the expression of Snail and E⁃cadherin，which are two well⁃characterized epithelial⁃mesenchymal transition（EMT）
markers. The results further showed that YAP regulated HCC cell migration and invasion by up ⁃ regulating EEF1A2. In addition，
CUT&RUN⁃qPCR analysis demonstrated YAP occupancy at the promoter of the EEF1A2 in HCC cells. Conclusion：YAP up⁃regulates
the expression of EEF1A2 through interacting with the sequence on the promoter of the EEF1A2，and subsequently promotes HCC cell
migration and invasion.

［基金项目］ 国家自然科学基金（81672336）；江苏省大学生创新创业训练计划项目（202011117056Y，202111117038Z）
∗
通信作者（Corresponding author），E⁃mail：shenwg@yzu.edu.cn