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


METHODS
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.


RESULTS
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.


CONCLUSIONS
PLC-gamma 1 may play an important role in metastasis and anti-apoptosis in human colorectal carcinomas.
Tan
Li
L
Department of Cell Biology, Southern Medical University, and Zhujiang Hospital, Guangzhou 510515, China.
Xiao
Bing-xiang
BX
Zeng
Wei-sen
WS
Lin
Jun
J
Zou
Zhi-peng
ZP
Xu
Ai-min
AM
Luo
Shen-qiu
SQ
D017209:Apoptosis / Q000187:drug effects; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000473:pathology / Q000628:therapy*; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000037:antagonists & inhibitors / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D051966:Phospholipase C gamma / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology; D034741:RNA, Small Interfering / Q000627:therapeutic use*
D007797:Laminin; D034741:RNA, Small Interfering; C084098:laminin A; D051966:Phospholipase C gamma; D005472:Fluorouracil