The effect of Fas gene transfection in combination with cisplatin on rectal carcinoma cells in vitro
Cai Hui-yu
Abstract
Cai Hui-yu
Abstract
Objective To study the effect of Fas gene transfection on rectal carcinoma cells in vitro. Methods By using RT-PCR technique, a full length of Fas gene 1007 bp was cloned from actived peripheral mononuclear cells of healthy donors. The fragment was ligated with the pGEM-T Easy and sequenced. The constructed vector was transfected into 8348 cells with lipofectin, the change in expression of Fas gene was determined by RT-PCR. The apoptosis and proliferation of rectal carcinoma cells pre- and posttransfection induced by cisplatin were analysed by ladder and MTT methods. Results Transfection of Fas gene significantly upregulates the expression of Fas in human rectal carcinoma 8348 cells. With the concentration of cisplatin at the level of 1, 5, 10, 20 and 40 mg/L , respectively, the suppression rates of Fas transfection group and control group were 47.2%51.8%57.2%65.4%71.0% and 29.6%33.0%37.8%41.4%47.0% respectively(t=15.33, P0.01).Conclusion Transfection of Fas gene by lipofectin significantly upregulates the expression of Fas in target cells. A strong suppression of rectal carcinoma cells was observed by combined Fas gene transfection and cisplatin.
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Objective To study the effect of Fas gene transfection on rectal carcinoma cells in vitro. Methods By using RT-PCR technique, a full length of Fas gene 1007 bp was cloned from actived peripheral mononuclear cells of healthy donors. The fragment was ligated with the pGEM-T Easy and sequenced. The constructed vector was transfected into 8348 cells with lipofectin, the change in expression of Fas gene was determined by RT-PCR. The apoptosis and proliferation of rectal carcinoma cells pre- and posttransfection induced by cisplatin were analysed by ladder and MTT methods. Results Transfection of Fas gene significantly upregulates the expression of Fas in human rectal carcinoma 8348 cells. With the concentration of cisplatin at the level of 1, 5, 10, 20 and 40 mg/L , respectively, the suppression rates of Fas transfection group and control group were 47.2%51.8%57.2%65.4%71.0% and 29.6%33.0%37.8%41.4%47.0% respectively(t=15.33, P0.01).Conclusion Transfection of Fas gene by lipofectin significantly upregulates the expression of Fas in target cells. A strong suppression of rectal carcinoma cells was observed by combined Fas gene transfection and cisplatin.
Key concepts: Transfection, Cisplatin, Apoptosis, Molecular biology, Peripheral blood mononuclear cell, In vitro, Medicine, Carcinoma