MicroRNA-125b attenuates epithelial-mesenchymal transitions and targets stem-like liver cancer cells through small mothers against decapentaplegic 2 and 4

Jun-Nian Zhou  ^{1   2} , Quan Zeng  ^{1   2} , Hai-Yang Wang  ^{1   2} , Biao Zhang  ^{1   2} , Si-Ting Li  ^{1   2} , Xue Nan  ^{1   2} , Ning Cao  ^{1   2} , Chun-Jiang Fu  ³ , Xin-Long Yan  ¹ , Ya-Li Jia  ^{1   2} , Jing-Xue Wang  ^{1   2} , Ai-Hua Zhao  ¹ , Zhi-Wei Li  ⁴ , Yan-Hua Li  ^{1   2} , Xiao-Yan Xie  ^{1   2} , Xiao-Mei Zhang  ⁵ , Yan Dong  ⁵ , Ying-Chen Xu  ⁶ , Li-Juan He  ^{1   2} , Wen Yue  ^{1   2} , Xue-Tao Pei  ^{1   2}

Affiliations

• ¹ Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing, China.
• ² South China Research Center for Stem Cell & Regenerative Medicine, AMMS, Guangzhou, China.
• ³ Fangshan Traditional Chinese Medicine Hospital, Beijing, China.
• ⁴ Department of Hepatobiliary Surgery, Beijing 302 Hospital, Beijing, China.
• ⁵ Department of Gastroenterology & Hepatology, Chinese PLA General Hospital, Beijing, China.
• ⁶ Department of Hepatobiliary Surgery, Beijing Tongren Hospital, Beijing, China.

Abstract

Emerging evidence suggests that epithelial-mesenchymal transitions (EMTs) play important roles in tumor metastasis and recurrence. Understanding molecular mechanisms that regulate the EMT process is crucial for improving treatment of hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) play important roles in HCC; however, the mechanisms by which miRNAs target the EMT and their therapeutic potential remains largely unknown. To better explore the roles of miRNAs in the EMT process, we established an EMT model in HCC cells by transforming growth factor beta 1 treatment and found that several tumor-related miRNAs were significantly decreased. Among these miRNAs, miR-125b expression was most strongly suppressed. We also found down-regulation of miR-125b in most HCC cells and clinical specimens, which correlated with cellular differentiation in HCC patients. We then demonstrated that miR-125b overexpression attenuated EMT phenotype in HCC cancer cells, whereas knockdown of miR-125b promoted the EMT phenotype in vitro and in vivo. Moreover, we found that miR-125b attenuated EMT-associated traits, including chemoresistance, migration, and stemness in HCC cells, and negatively correlated with EMT and cancer stem cell (CSC) marker expressions in HCC specimens. miR-125b overexpression could inhibit CSC generation and decrease tumor incidence in the mouse xenograft model. Mechanistically, our data revealed that miR-125b suppressed EMT and EMT-associated traits of HCC cells by targeting small mothers against decapentaplegic (SMAD)2 and 4. Most important, the therapeutic delivery of synthetic miR-125b mimics decreased the target molecule of CSC and inhibited metastasis in the mice model. These findings suggest a potential therapeutic treatment of miR-125b for liver cancer.

Conclusion: miR-125b exerts inhibitory effects on EMT and EMT-associated traits in HCC by SMAD2 and 4. Ectopic expression of miR-125b provides a promising strategy to treat HCC.