title: Lentiviral vector-mediated doxycycline-inducible USP39 shRNA or cDNA expression in triple-negative breast cancer cells.
abstract: Triple-negative breast cancer (TNBC), characterized by distinct biological and clinicopathological features, has a poor prognosis due to lack of effective therapeutic targets. Our previous data revealed that high levels of USP39 were selectively present in TNBC samples compared with their normal breast tissue samples and USP39 was also expressed at different levels in cultured TNBC cells and normal breast cells. Yet, the underlying cellular and molecular mechanisms of USP39 remain unclear. In the present study, we describe a doxycycline (DOX)-regulated lentiviral vector system expressing shRNA or cDNA of the USP39 gene in the TNBC cell line MDA-MB-231. USP39 expression was knocked down by the miR-30-based inducible lentiviral short hairpin RNA (shRNA) delivery system or overexpressed by the inducible cDNA system. The inducible shRNA-mediated downregulation of USP39 expression markedly reduced the proliferation and colony-forming ability of MDA-MB-231 cells, while overexpression of USP39 by the inducible system did not promote cancer cell proliferation. The lentiviral vector-mediated Tet-on system demonstrated efficient and inducible knockdown of USP39 or overexpression of USP39 in TNBC cells, facilitating a wide variety of applications for gene knockdown and overexpression experiments in gene functional studies in vitro and in vivo.
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Shihai
authors: S
authors: Liu
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Xiangping
authors: X
authors: Liu
authors: Center of Diagnosis and Treatment of Breast Disease, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Haibo
authors: H
authors: Wang
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Quan
authors: Q
authors: Zhou
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Ye
authors: Y
authors: Liang
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Aihua
authors: A
authors: Sui
authors: Medical Research Center, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Ruyong
authors: R
authors: Yao
authors: Center of Diagnosis and Treatment of Breast Disease, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Bin
authors: B
authors: Zhao
authors: Center of Diagnosis and Treatment of Breast Disease, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
authors: Ming
authors: M
authors: Sun
mesh_terms: D001940:Breast / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D004318:Doxycycline / Q000494:pharmacology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D055785:Gene Knockdown Techniques / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D064726:Triple Negative Breast Neoplasms / Q000235:genetics*; D064570:Ubiquitin-Specific Proteases / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D035683:MicroRNAs; D034741:RNA, Small Interfering; C579218:USP39 protein, human; D064570:Ubiquitin-Specific Proteases; D004318:Doxycycline