title: Reciprocal activation between STAT3 and miR-181b regulates the proliferation of esophageal cancer stem-like cells via the CYLD pathway.
abstract: Recent 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.
  
  
  
  Serum-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.
  
  
  
  Sphere 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.
  
  
  
  The 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.
authors.affiliation: 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.
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Xu
authors.affiliation: Department of Pathogen Biology and Immunology, School of Basic Course, Guangdong Pharmaceutical University, Guangzhou, 510006, P.R. China.
authors.forename: Peng-Jun
authors.initials: PJ
authors.lastname: Zhou
authors.affiliation: 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.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou, 510006, P.R. China.
authors.forename: Wu-Yu
authors.initials: WY
authors.lastname: Fu
authors.affiliation: School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou, 510006, P.R. China.
authors.forename: Bi-Bo
authors.initials: BB
authors.lastname: Ruan
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Hai-Peng
authors.initials: HP
authors.lastname: Xu
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Chao-Zhi
authors.initials: CZ
authors.lastname: Hu
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Tian
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Qin
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Li
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Qiu-Ying
authors.initials: QY
authors.lastname: Liu
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Ren
authors.affiliation: 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. zhangrong@sysucc.org.cn.
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.affiliation: College of Life Science and Technology, Jinan University, 510632, Guangzhou, P.R. China. twang-yf@163.com.|Key laboratory of Bioengineering medicine of Guangdong Province, Jinan University, 510632, Guangzhou, P.R. China. twang-yf@163.com.
authors.forename: Yi-Fei
authors.initials: YF
authors.lastname: Wang
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001665:Binding Sites; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D000074749:Deubiquitinating Enzyme CYLD; D004195:Disease Models, Animal; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D034622:RNA Interference; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
chemical_list: D020413:3' Untranslated Regions; C000608460:MIRN-181 microRNA, human; D035683:MicroRNAs; D050796:STAT3 Transcription Factor; D025521:Tumor Suppressor Proteins; C409984:CYLD protein, human; D000074749:Deubiquitinating Enzyme CYLD
keywords: CYLD; Esophageal cancer stem-like cells; Proliferation; STAT3; Sphere formation cells; miR-181b