TCRVβ基因亚家族的优势取用和PTK信号传导途径的激活及体外对肝癌细胞凋亡的诱导

摘要

Objective: To study the expression of TCRVβ subfamily which specially recognize the hepatoma cell antigen and the apoptosis of hepatoma cell induced by McAb costimulated PBLs.Methods: The change of the phenotype of PBLs was studied by flow cell cytometry and the level of the expression of TCRVβ was studied by RT-PCR and Southern blot,the PTX by westem blot. The hypermicroscopic ultrastructure was ob served through transmission electron microscope. Results: The level of CD3 and CD8 of PBLs was significantly increased after acted with hepatoma cells, while there was no change in CD4. The expression of TCRVβ7of PBLs was dramaticly increased and peaked at 4 days. PTK in creased correspondently, to 58% compared with 11% in control. Besides anti-CD3 McAb induced lymphocyte apoptosis, the mediated apoptosis of hepatoma cells was found in the other three groups. Conclusion: TCRVβ7was the tumor antigen specific T cell receptor, and it activate the PTK signal pathway.The McAb activated lymphocytes initiated apoptosis in hepatoma cells.%目的：研究特异识别肝癌细胞株肿瘤相关抗原的TCRVβ基因亚家族的优势取用及对肝癌细胞凋亡的诱导。方法：流式细胞术检测淋巴细胞表型，RT-PCR和Southem印迹分析TCRVβ基因亚家族表达水平，Western blot检测PIX含量。透射电镜观察凋亡细胞超微结构。结果：McAb共刺激T细胞与肝癌细胞混合培养后，T细胞表面CD3和CD8分子表达量明显升高，而CD4无明显变化。TCRVβ4选择性扩增，表达水平由5％升高至13％～25％，且在第4天达到高峰。同时相应的PTK信号传导途径被激活，其含量由11％升至58％，亦在第4,5天达到高峰。以抗CD3+CD28，抗CD28+CD80，抗CD2+CD58共刺激的淋巴细胞均在体外诱导了肝癌细胞的凋亡。结论：TCRVβ7为特异的肿瘤抗原识别受体，TCR-CD4复合物与抗原结合后激活PTK信号传导途径，并诱导了肝癌细胞的凋亡。