TCR Vβ7基因转染PBLs的表达及抗肝癌的作用

Expression and anti-hepatoma effect of TCR Vβ7 after being transfected into PBLs

目的 :将TCRVβ7基因转化到淋巴细胞后研究肝癌细胞的生物学作用。 方法 :用RT PCR的方法扩增出TCRVβ7基因 ,克隆到表达载体pLXSN上 ,用脂质体转染的方法 ,将重组载体导入淋巴细胞 ,然后将淋巴细胞作用于肝癌细胞 ,淋巴细胞用流式细胞仪测表型和用激光共聚焦检测TCRVβ7 1基因的表达 ,肝癌细胞作超微结构分析。 结果 :TCRVβ7基因表达的跨膜蛋白显著增多 ,而且淋巴细胞增多 (P <0 0 5 ) ,肝癌细胞出现凋亡。结论 :TCRVβ7亚家族具有识别肝癌抗原的作用并能激活T细胞的杀伤肝癌细胞的功能。

Objective:To study the biological effects of TCR on hepatoma cells by transfecting Vβ7 into lymphocytes.Methods:TCR Vβ7 gene was amplified by RT PCR and cloned to expression vector pLXSN. The recombinant was transferred into lymphocytes by Lipofectin Reagent transfection, then the lymphocytes were co cultured with hepatoma cells. The phenotype of lymphocytes was detected on the Flow Cytometry, the expression of TCR Vβ7.1 gene was detected by Laser Scanning Confocal Microscope ( LSCM) and the ultrastructure of the hepatoma cells was showed by electronic microscope.Results:The amount of the transmembrane protein expressed by TCR Vβ7.1 gene was increased significantly, and so was the amount of lymphocytes (P<0 05). Apoptosis appeared in the hepatoma cells. Conclusion:TCR Vβ7 subfamily can recognize hepatoma antigen and activate T cell to kill hepatoma cells. [