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Background In most colorectal carcinomas,the level of phospholipase C (PLC)-gamma 1 expression is greatlyelevated.Increased expression of PLC-gamma 1 may play an important role in colon carcinogenesis,but the mechanismis not well known.The aim of this study was to evaluate the role of PLC-gamma 1 in colon carcinogenesis by usingrecombinant 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 byeach lentivirus,stably transduced cells were selected by Blasticidin.The protein and mRNA expression of PLC-gamma 1were examined by Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis,and the effectsof 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 withoutaffecting the levels of other subtypes of PLC so that the role of PLC-gamma 1 in colon carcinogenesis could be examinedSilencing of endogenous PLC-gamma 1 resulted in efficient inhibition of the adhesion and migration of LoVo cells in vitroand 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 colorectalcarcinomas.
Background In most colorectal carcinomas, the level of phospholipase C (PLC) -gamma 1 expression is greatly enhanced. 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 lentivirus , stably transduced cells were selected by Blasticidin. The protein and mRNA expression of PLC-gamma 1were 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.