【摘要】 为了探讨肝癌细胞反击肿瘤浸润淋巴细胞(TIL)的机制,在体外进行肝癌细胞和TIL混合培养后,检测两种细胞FasL、Fas、caspase-8基因的表达情况,以及肿瘤细胞反击时TIL凋亡比例的变化。将肝癌细胞与TIL按照不同的比例共培养后,流式细胞术检测TIL凋亡率;实时荧光定量PCR检测肝癌细胞与TILFasL、Fas和caspase-8基因的表达情况;以及Western印迹检测FasL、caspase-8的表达情况。不同浓度的肝癌细胞与TIL共同培养48h后,随着肝癌细胞接种浓度的增加,TIL凋亡率明显增加(P<0.01)。与正常人肝细胞相比,人肝癌细胞FasLmRNA表达含量明显增高(P<0.01)。与人肝癌细胞共同培养24h后,TILFas、caspase-8基因mRNA的表达也明显升高;TILcaspase-8的表达也明显升高。结果表明,肝癌细胞可以通过Fas系统诱导TIL发生凋亡,这为肝癌的免疫逃逸和肿瘤反击机制提供了依据。更多还原

【Abstract】 To investigate the mechanisms of hepatic cancer cells inducing apoptosis of tumor-infiltrating lymphocytes (TILs) in vitro, hepatic cancer cells were co-cultured with TILs in different effector-taget ratio. Apoptotic ratios of TILs were screened by flow cytometry (FCM). Expression levels of mRNA of FasL, Fas and caspase-8 in hepatic cancer cells and TILs were tested by real time PCR. FasL and caspase-8 expression were screened by immunoblotting. After co-culturing with hepatic cancer cells, apoptotic ratios of TILs were increased with the ascending number of hepatic cancer cells (P<0.01). Compared with the normal hepatic cells from healthy donors, FasL mRNA expression level of hepatic cancer cells was significantly increased (P<0.01). After 24 hour’s co-culturing with hepatic cancer cells, Fas and caspase-8 mRNA expression levels of TILs were significantly higher than before co-culturing (P<0.05). After co-culturing with hepatic cancer cells, caspase-8 expression of TILs was significantly higher than before. The results suggested that hepatic cancer cells could induce apoptosis of TILs through Fas system, providing a evidence for the metastasis and counterattack of hepatic cancer cells.更多还原