Y Chen1, W Hu, Y Lu, S Jiang, C Li, J Chen, D Tao, Y Liu, Y Yang, Y Ma. 1. Department of Medical Genetics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610000, China.
Abstract
OBJECTIVES: PIWIL2 is widely expressed in various tumours and implicated in playing a role in tumourigenesis. For a more thorough study of PIWIL2 functions in tumour cells, we aimed to establish PIWIL2-specific transcript knock-down/knockout HepG2 cell lines using transcription activator-like effector nuclease (TALEN) technology. Furthermore, we proposed to use the cell models to explore PIWIL2 functions in TGF-β signalling in HepG2 cells. HepG2s are human hepatocellular carcinoma cells. MATERIALS AND METHODS: We established PIWIL2 knock-down or knock-out cell lines in HepG2 using TALEN technology. Next, we sought to use the cell line models to investigate effects of full length PIWIL2-specific transcripts in cell proliferation induced by TGF-β. RESULTS: First, we established PIWIL2-specific transcript mono-allele and bi-allele knockout HepG2 cell lines. By using the cell line models, we found that specific transcript knockdown of full length PIWIL2 can suppress cell proliferation, while ectopic expression of PIWIL2 enhanced proliferation of HepG2 by suppressing the TGF-β pathway. Furthermore, we demonstrated that PIWIL2 can interact with HSP90 to prevent formation of HSP90-TβR complexes, which promote degradation of TβRs. CONCLUSIONS: Taken together, our study revealed critical negative regulation of TGF-β signalling by PIWIL2 in HepG2 tumour cells, and provided an effective strategy to study specific gene transcript functions in cells.
OBJECTIVES:PIWIL2 is widely expressed in various tumours and implicated in playing a role in tumourigenesis. For a more thorough study of PIWIL2 functions in tumour cells, we aimed to establish PIWIL2-specific transcript knock-down/knockout HepG2 cell lines using transcription activator-like effector nuclease (TALEN) technology. Furthermore, we proposed to use the cell models to explore PIWIL2 functions in TGF-β signalling in HepG2 cells. HepG2s are human hepatocellular carcinoma cells. MATERIALS AND METHODS: We established PIWIL2 knock-down or knock-out cell lines in HepG2 using TALEN technology. Next, we sought to use the cell line models to investigate effects of full length PIWIL2-specific transcripts in cell proliferation induced by TGF-β. RESULTS: First, we established PIWIL2-specific transcript mono-allele and bi-allele knockout HepG2 cell lines. By using the cell line models, we found that specific transcript knockdown of full length PIWIL2 can suppress cell proliferation, while ectopic expression of PIWIL2 enhanced proliferation of HepG2 by suppressing the TGF-β pathway. Furthermore, we demonstrated that PIWIL2 can interact with HSP90 to prevent formation of HSP90-TβR complexes, which promote degradation of TβRs. CONCLUSIONS: Taken together, our study revealed critical negative regulation of TGF-β signalling by PIWIL2 in HepG2 tumour cells, and provided an effective strategy to study specific gene transcript functions in cells.
Authors: James J Liu; Rulong Shen; Li Chen; Yin Ye; Gang He; Keding Hua; David Jarjoura; Toru Nakano; Ganju K Ramesh; Charles L Shapiro; Sanford H Barsky; Jian-Xin Gao Journal: Int J Clin Exp Pathol Date: 2010-03-20
Authors: Yin Ye; De-Tao Yin; Li Chen; Quansheng Zhou; Rulong Shen; Gang He; Qingtao Yan; Zhenyu Tong; Andrew C Issekutz; Charles L Shapiro; Sanford H Barsky; Haifan Lin; Jian-Jian Li; Jian-Xin Gao Journal: PLoS One Date: 2010-10-20 Impact factor: 3.240
Authors: Xiaoxin Mu; Shu Lin; Junhua Yang; Chen Chen; Yun Chen; Maryanne C Herzig; Kenneth Washburn; Glenn A Halff; Christi A Walter; Beicheng Sun; Lu-Zhe Sun Journal: PLoS One Date: 2013-05-21 Impact factor: 3.240