[Roles of hMMS2 gene in reversing the oxaliplatin tolerance of human colon carcinoma cells].
In this study, the roles of hMMS2 (human methyl methanesulfonate sensitive mutant 2) gene encoding the human ubiquitin-conjugating enzyme E2 variant 2 in the drug resistance in human colon carcinoma were investigated by using a well-differentiated human colorectal carcinoma L-OHP-resistant cell line, THC8307/L-OHP. THC8307/L-OHP cells were transfected via liposome along with plasmid pcDNA6.2-GW/EmGFP-miR-MMS2 expressing both miRNA against hMMS2 and GFP, followed by real-time fluorescent quantitative PCR and immunofluorescence to select stable transfectants with significantly reduced hMMS2 expression. Compared with untransfected or pcDNA6.2-GW/EmGFP vector-transfected cells, the hMMS2-depleted cells displayed significantly (P<0.05) reduced half inhibition concentration(IC50) resistance index (RI) and colony-forming efficiency (CFE) upon treatment with oxaliplatin (L-OHP), while its relative reverse efficiency(RRE) was significantly higher (P<0.05) than the control cells, indicating compromised ability of cell proliferation. Indeed, Rho-damine 123 staining and flow cytometry analyses revealed an increased rate of apoptosis in hMMS2-depleted cells while no difference in cell proliferation or apoptosis was observed between the two control cell lines. The above observations collec-tively indicate that suppression of hMMS2 reverses L-OHP tolerance in differentiated human colorectal carcinoma cells by promoting apoptosis.
Ningxia Key Laboratory of Reproduction and Genetics, College of Inspection, Ningxia Medical University, Yinchuan 750004, China.
Lei
L
Zhang
Center for Translational Medicine, School of Medicine, Tongji University, Shanghai 200072, China.
Yu
Y
Sui
Ningxia Key Laboratory of Reproduction and Genetics, College of Inspection, Ningxia Medical University, Yinchuan 750004, China.
Ting
T
Wang
Ningxia Key Laboratory of Reproduction and Genetics, College of Inspection, Ningxia Medical University, Yinchuan 750004, China.
Lijian
L
Li
Ningxia Key Laboratory of Reproduction and Genetics, College of Inspection, Ningxia Medical University, Yinchuan 750004, China.
Yuanjie
Y
Li
Center for Translational Medicine, School of Medicine, Tongji University, Shanghai 200072, China.
Caixia
C
Jin
Ningxia Key Laboratory of Reproduction and Genetics, College of Inspection, Ningxia Medical University, Yinchuan 750004, China.
Fang
F
Xu
D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing; D006801:Humans; D008025:Ligases / Q000172:deficiency / Q000235:genetics*; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D034741:RNA, Small Interfering / Q000235:genetics; D017423:Sequence Analysis, RNA; D044763:Ubiquitin-Conjugating Enzymes
D009944:Organoplatinum Compounds; D034741:RNA, Small Interfering; D000077150:Oxaliplatin; C114715:UBE2V2 protein, human; D044763:Ubiquitin-Conjugating Enzymes; D008025:Ligases