Fas转染及抗体处理对舌鳞癌细胞Tca8113移植瘤形成和增殖的影响

Effect of Fas Gene Transfection on Tumourigenicity and Proliferation of Transplanted Tumor of Oral Squamous Cell Carcinoma Cell Line Tca8113

目的 研究Fas转染和抗Fas单克隆抗体对舌鳞癌细胞系Tca8113裸鼠体内移植瘤形成和增殖的影响 ,探讨相关机制。方法 脂质体法以真核表达重组质粒pBK_fas转染舌鳞癌细胞系Tca8113。部分转染细胞行抗Fas单克隆抗体处理。未转染细胞、Fas转染细胞、Fas转染抗体处理细胞分别接种裸鼠皮下。观测移植瘤生长 ,绘制生长曲线。RT_PCR检测肿瘤细胞FasmRNA表达。流式细胞术 (FCM)检测肿瘤细胞凋亡、增殖及Fas蛋白表达。结果 Fas转染和抗体处理延迟肿瘤形成 ,抑制肿瘤增殖。Fas转染上调移植瘤FasmRNA表达 ,提高Fas蛋白表达强度和凋亡指数 ,但不影响Fas蛋白阳性表达率和增殖指数。抗体处理不影响FasmRNA和蛋白表达 ,但可提高凋亡指数 ,降低增殖指数。结论 Fas转染抑瘤效应是上调Fas转录和表达、促进凋亡的结果。抗Fas单克隆抗体抑瘤效应则与激活凋亡和抑制增殖有关。

Objective To investigate the effect of fas gene transfection and monoclonal anti-fas antibody on tumorigenicity and proliferation of transplanted tumor of Tca8113 cell.Methods Plasmid including fas gene was transfected into Tca8113 cell by lipofectamine kit. Some transfected cells were treated by monoclonal anti-fas antibody after 48 hours since transfection. Untransfected cell (control), fas-tansfected cell and fas-transfected cell treated with antibody were transplanted to nude mice subcutaneously. Growth of transplanted tumor was observed and recorded regularly. Animals were sacrificed and tumor samples were harvested at the end of experiment. Fas expression in each neoplasm was assessed by RT-PCR. Apoptosis, proliferation and expression of fas protein in tumor tissue were measured by flow cytometry (FCM).Results Tumor occurred much later in fas-transfected group and fas-transfected plus antibody treated group. Growth arrest was found in them. RT-PCR and FCM suggested that fas-transfection up-regulated the expression of fas mRNA and protein, increased apoptosis index (AI). But no effect on proliferation index (PI) was observed. Monoclonal anti-fas antibody did not effect the expression of fas mRNA and protein, but increased AI and decreased PI.Conclusion Fas-transfection suppressing tumorigenesis of Tca8113 cell transplanted in nude mice might be caused by up-regulation of expression of fas gene and enhancement of apoptosis. However, anti-fas antibody suppressing tumorigenesis might be associated with activation of apoptosis and repression of proliferation.