Open AccessA TALEN ‐based specific transcript knock‐down of PIWIL 2 suppresses cell growth in HepG2 tumor cell
Author(s) -
Chen Y.,
Hu W.,
Lu Y.,
Jiang S.,
Li C.,
Chen J.,
Tao D.,
Liu Y.,
Yang Y.,
Ma Y.
Publication year - 2014
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/cpr.12120
Subject(s) - transcription activator like effector nuclease , gene knockdown , biology , cell growth , ectopic expression , microbiology and biotechnology , cell culture , gene knockout , cell , nuclease , effector , gene , genetics , genome editing , crispr
Objectives PIWIL 2 is widely expressed in various tumours and implicated in playing a role in tumourigenesis. For a more thorough study of PIWIL 2 functions in tumour cells, we aimed to establish PIWIL 2‐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 PIWIL 2 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 PIWIL 2‐specific transcripts in cell proliferation induced by TGF ‐β. Results First, we established PIWIL 2‐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 PIWIL 2 can suppress cell proliferation, while ectopic expression of PIWIL 2 enhanced proliferation of HepG2 by suppressing the TGF ‐β pathway. Furthermore, we demonstrated that PIWIL 2 can interact with HSP 90 to prevent formation of HSP 90–TβR complexes, which promote degradation of TβRs. Conclusions Taken together, our study revealed critical negative regulation of TGF ‐β signalling by PIWIL 2 in HepG2 tumour cells, and provided an effective strategy to study specific gene transcript functions in cells.