Reciprocal activation between STAT3 and miR-181b regulates the proliferation of esophageal cancer stem-like cells via the CYLD pathway

Molecular Cancer

Research

Chao-zhi Hu¹  Xiao Wang¹  Hai-peng Xu¹  Lu Tian¹  Jin-hong Qin¹  Qiu-ying Liu¹  Sheng Wang¹  Li Zhang¹  Yi-cheng Li¹  Zhe Ren¹  Ying Wang²  Dan-dan Xu³  Yi-fei Wang³  Peng-jun Zhou⁴  Wu-yu Fu⁵  Bi-bo Ruan⁵  Rong Zhang⁶ 

[1] College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China;College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China;Faculty of Environmental and Biological Engineering, Guangdong University of Petrochemical Technology, 525000, Maoming, P.R. China;College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China;Key laboratory of Bioengineering medicine of Guangdong Province, Jinan University, 510632, Guangzhou, P.R. China;Department of Pathogen Biology and Immunology, School of Basic Course, Guangdong Pharmaceutical University, 510006, Guangzhou, P.R. China;School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, 510006, Guangzhou, P.R. China;State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, 510632, Guangzhou, P.R. China;

关键词: Esophageal cancer stem-like cells;    Sphere formation cells;    STAT3;    miR-181b;    Proliferation;    CYLD;   

DOI  :  10.1186/s12943-016-0521-7

 received in 2016-01-19, accepted in 2016-05-05,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundRecent studies have suggested that cancer cells contain subpopulations that can initiate tumor growth, self-renew, and maintain tumor cell growth. However, for esophageal cancer cells, the relationship between STAT3, microRNAs and cancer stem cells remains unclear.MethodsSerum-free culture was used to enrich esophageal cancer stem-like cells (ECSLC). Flow cytometry determined the proportion of ECSLC. qPCR were performed to examine expression level of stemness factors, mesenchymal markers, ATP-binding cassette (ABC) transporters, STAT3, miR-181b, CYLD. Western blot were performed to analyze the expression of STAT3, p-STAT3 and CYLD (cylindromatosis). BALB/c mice xenograft studies were conducted to evaluate the tumorigenicity of enriched ECSLC. Sphere formation assay and colony formation assays were employed to analyze the relationship between STAT3 and miR-181b. Luciferase assays were used to evaluate activity which CYLD is a target of miR-181b.ResultsSphere formation cells (SFCs) with properties of ECSLC were enriched. Enriched SFCs in serum-free suspension culture exhibited cancer stem-like cell properties and increased single-positive CD44 + CD24-, stemness factor, mesenchymal marker expression ABC transporters and tumorigenicity in vivo compared with the parental cells. Additionally, we found that reciprocal activation between STAT3 and miR-181b regulated SFCs proliferation. Moreover, STAT3 directly activated miR-181b transcription in SFCs and miR-181b then potentiated p-STAT3 activity. Luciferase assays indicated that CYLD was a direct and functional target of miR-181b.ConclusionThe mutual regulation between STAT3 and miR-181b in SFCs was required for proliferation and apoptosis resistance. STAT3 and miR-181b control each other’s expression in a positive feedback loop that regulates SFCs via CYLD pathway. These findings maybe is helpful for targeting ECSLC and providing approach for esophageal cancer treatments.

【 授权许可】

CC BY   
© Xu et al. 2016