Construction of lentivirus producing PLC-gamma1 siRNA and its effect on apoptosis of human colorectal carcinomas cell lines.
- Author:
Li TAN
1
;
Shen-qiu LUO
;
Jun LIN
Author Information
- Publication Type:Journal Article
- MeSH: Antimetabolites, Antineoplastic; pharmacology; Apoptosis; drug effects; genetics; Blotting, Western; Cell Line, Tumor; Colorectal Neoplasms; genetics; pathology; DNA, Recombinant; genetics; Fluorouracil; pharmacology; Genetic Vectors; Humans; Lentivirus; genetics; Phospholipase C gamma; genetics; metabolism; RNA Interference; RNA, Messenger; biosynthesis; genetics; RNA, Small Interfering; genetics; Reverse Transcriptase Polymerase Chain Reaction; Transfection
- From: Journal of Southern Medical University 2007;27(3):255-258
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct recombinant lentivirus that stably suppresses phospholipase C (PLC) gamma1 expression in human colorectal carcinoma LoVo cells to obtain LoVo cell lines deficient in PLC gamma1 for investigation of the role of PLC gamma1 gene.
METHODSRecombinant lentivirus producing PLC gamma1 siRNA were constructed to infect LoVo cells, and the stably transduced cells were selected with blasticidin. The protein and mRNA expression of PLC gamma1 was examined by Western blotting and RT-PCR, and the effect of the lentivirus on cell apoptosis was analyzed by flow cytometry.
RESULTS AND CONCLUSIONPLC gamma1 siRNA significantly suppressed PLC gamma1 expression in LoVo cells, suggesting high efficiency of gene silencing induced by the siRNA produced by the recombinant lentivirus. Concomitantly, cell apoptosis induced by 5-FU was significantly increased.
