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

• Published in: Chinese medical journal Vol. 120; no. 9; pp. 749 - 754
• Main Authors: Tan, Li, Xiao, Bing-xiang, Zeng, Wei-sen, Lin, Jun, Zou, Zhi-peng, Xu, Ai-min, Luo, Shen-qiu
• Format: Journal Article
• Language: English
• Published: China Department of Cell Biology, Southern Medical University, Guangzhou 510515, China%Department of Cell Biology,Zhujiang Hospital, Southern Medical University, Guangzhou 510515, China%Graduate School of Medicine, Hong Kong University, Hong Kong 999077, China 05-05-2007
• 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; migration; apoptosis; siRNA; adhesion; phospholipase C gamma; lentivirus
• ISSN: 0366-6999
• DOI: 10.1097/00029330-200705010-00003

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.