title: Downregulation of GRHL2 inhibits the proliferation of colorectal cancer cells by targeting ZEB1.
abstract: Previous reports have associated GRHL2 with tumor progression. However, the biological role of GRHL2 in human colorectal cancer (CRC) has not been explored. We examined the expression of GRHL2 in 75 CRC samples, as well as the paired non-tumor tissues, by immunohistochemistry, qRT-PCR, and western blot analysis. The association between GRHL2 expression and various clinicopathological parameters including Ki-67, a marker of proliferative activity, was also evaluated. We performed lentivirus-mediated shRNA transfection to knock down GRHL2 gene expression in HT29 and HCT116 CRC cells. Cell proliferation was examined by the CCK-8 (Cell Counting Kit-8) assay, colony formation, and cell cycle assay in vitro. Tumorigenesis in vivo was assessed using a mouse xenograft model. Moreover, we transiently silenced ZEB1 expression in GRHL2-knockdown CRC cells using specific shRNA, and then examined the effects on GRHL2 and E-cadherin expression, as well as cell proliferation. Herein, we demonstrated that enhanced GRHL2 expression was detected in CRC, and correlated with higher levels of Ki-67 staining, larger tumor size, and advanced clinical stage. Knocking down GRHL2 in HT29 and HCT116 CRC cells significantly inhibited cell proliferation by decreasing the number of cells in S phase and increasing that in the G 0/G 1 phaseof the cell cycle. This resulted in inhibition of tumorigenesis in vivo, as well as increased expression of ZEB1. Furthermore, transient ZEB1 knockdown dramatically enhanced cell proliferation and increased GRHL2 and E-cadherin expression. Collectively, our study has identified ZEB1 as a target of GRHL2 and suggested a reciprocal GRHL2-ZEB1 repressive relationship, providing a novel mechanism through which proliferation may be modulated in CRC cells.
authors.affiliation: Department of Surgery; Ruijin Hospital; Shanghai Jiao Tong University School of Medicine; Shanghai, PR China; Shanghai Institute of Digestive Surgery; Shanghai, PR China.
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Quan
authors.affiliation: Department of Surgery; Ruijin Hospital; Shanghai Jiao Tong University School of Medicine; Shanghai, PR China.
authors.forename: Runsen
authors.initials: R
authors.lastname: Jin
authors.affiliation: Shanghai Institute of Digestive Surgery; Shanghai, PR China.
authors.forename: Ao
authors.initials: A
authors.lastname: Huang
authors.affiliation: Shanghai Institute of Digestive Surgery; Shanghai, PR China.
authors.forename: Hongchao
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Surgery; Ruijin Hospital; Shanghai Jiao Tong University School of Medicine; Shanghai, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Feng
authors.affiliation: Department of Surgery; Ruijin Hospital; Shanghai Jiao Tong University School of Medicine; Shanghai, PR China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zang
authors.affiliation: Department of Surgery; Ruijin Hospital; Shanghai Jiao Tong University School of Medicine; Shanghai, PR China; Shanghai Minhang District Central Hospital; Shanghai, PR China.
authors.forename: Minhua
authors.initials: M
authors.lastname: Zheng
mesh_terms: D000368:Aged; D000818:Animals; D015820:Cadherins / Q000235:genetics; D002453:Cell Cycle; D049109:Cell Proliferation; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018450:Disease Progression; D015536:Down-Regulation; D005260:Female; D045325:HCT116 Cells; D019073:HT29 Cells; D064593:Heterografts; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D009368:Neoplasm Transplantation; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D015820:Cadherins; D004268:DNA-Binding Proteins; C470830:GRHL2 protein, human; D018398:Homeodomain Proteins; D014157:Transcription Factors; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1
keywords: E-cadherin; GRHL2; ZEB1; colorectal cancer; progression; proliferation; tumorigenesis
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