Antitumour effects on human colorectal carcinomas cells by stable silencing of phospholipase C-gamma 1 with lentivirus-delivered siRNA

In most colorectal carcinomas, the level of phospholipase C (PLC)-gamma 1 expression is greatly elevated. Increased expression of PLC-gamma 1 may play an important role in colon carcinogenesis, but the mechanism is not well known. The aim of this study was to evaluate the role of PLC-gamma 1 in colo...

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Published in:Chinese medical journal 2007-05, Vol.120 (9), p.749-754
Main Authors: Tan, Li, Xiao, Bing-xiang, Zeng, Wei-sen, Lin, Jun, Zou, Zhi-peng, Xu, Ai-min, Luo, Shen-qiu
Format: Article
Language:English
Subjects:
Apoptosis - drug effects
Cell Adhesion
Cell Line, Tumor
Colorectal Neoplasms - pathology
Colorectal Neoplasms - therapy
Fluorouracil - pharmacology
Humans
Laminin - antagonists & inhibitors
Laminin - genetics
Lentivirus - genetics
Phospholipase C gamma - antagonists & inhibitors
Phospholipase C gamma - genetics
Phospholipase C gamma - physiology
RNA, Small Interfering - therapeutic use
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Summary:In most colorectal carcinomas, the level of phospholipase C (PLC)-gamma 1 expression is greatly elevated. Increased expression of PLC-gamma 1 may play an important role in colon carcinogenesis, but the mechanism is not well known. The aim of this study was to evaluate the role of PLC-gamma 1 in colon carcinogenesis by using recombinant lentivirus that stably suppressed the PLC-gamma 1 expression in human colorectal carcinoma LoVo cells. Recombinant lentivirus producing PLC-gamma 1 siRNA were prepared. After LoVo cells were transduced by each lentivirus, stably transduced cells were selected by Blasticidin. The protein and mRNA expression of PLC-gamma 1 were examined by Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis, and the effects of the lentivirus on the cell adhesion, migration and apoptosis were analyzed. Stable LoVo cell line deficient in PLC-gamma 1, was established. Notably, PLC-gamma 1 was silenced without affecting the levels of other subtypes of PLC so that the role of PLC-gamma 1 in colon carcinogenesis could be examined. Silencing of endogenous PLC-gamma 1 resulted in efficient inhibition of the adhesion and migration of LoVo cells in vitro and a great increase of 5-fluorouracil induced apoptosis (30%-40%) of LoVo cells. PLC-gamma 1 may play an important role in metastasis and anti-apoptosis in human colorectal carcinomas.
ISSN:0366-6999
DOI:10.1097/00029330-200705010-00003