sFasL及其受体在双阴性T细胞杀伤胰腺癌细胞中的作用及意义

The role and significance of sFasL and its receptor in killing pancreatic cancer cells by double negative T cells

目的探讨s Fas L(CD178)及其受体Fas(CD95,APO-1)在双阴性T细胞(DNT cell)杀伤胰腺癌细胞中的作用及意义。方法采用OKT3抗体吸附法培养DNT细胞。ELISA法检测DNT细胞上清液中的s Fas L的含量。Western blot和qRT-PCR法检测3株胰腺癌细胞中Fas的表达量。免疫组化法检测60例胰腺癌组织中Fas的表达情况,并分析其与临床病理参数之间的关系。结果 ELISA法显示DNT细胞上清液中s Fas L含量比对照组高(t=3.00,P<0.01)。Western blot法检测Fas在3株胰腺癌细胞中均有较高表达,从低到高依次为Capan-1、BXPC-3、Panc-1。qRT-PCR法检测Fas mRNA在3株胰腺癌细胞中均有表达,从低到高依次为Capan-1、BXPC-3、Panc-1,且两两之间的差异均有统计学意义(F=211.56,P<0.05)。免疫组化法检测Fas在癌旁组织的阳性表达率高于癌组织(χ~2=8.62,P<0.05)。结论s Fas L/Fas途径可能是DNT细胞杀伤胰腺癌细胞的机制之一。

Objective To investigate the role and significance of sFasL(CD178)and its receptor Fas(CD95/APO-1)in killing pancreatic cancer cells by double negative T cells(DNT cells).Methods DNT cells were cultured by OKT3 antibody adsorption method.ELISA method was used to detect the content of sFasL in DNT cell supernatant.Western blot and qRT-PCR were used to detect the expression of Fas in three pancreatic cancer cells.Immunohistochemistry was used to detect the expression of Fas in 60 cases of pancreatic cancer and its relationship with clinicopathological parameters was analyzed.Results ELISA method showed that the content of sFasL in the supernatant of DNT cells was higher than that of the control group(t=3.00,P<0.01).The results of Western blot suggested that Fas was highly expressed in all three pancreatic cancer cells,ranging from low to high were Capan-1,BXPC-3 and Panc-1.The expression of Fas mRNA was detected by qRT-PCR in all three pancreatic cancer cells,and the sequence was Capan-1,BXPC-3 and Panc-1 from low to high(F=211.56,P<0.05).Immunohistochemistry results showed that the expression of Fas was decreased in pancreatic tissue compared with paracancerous tissue(χ2=8.62,P<0.05).Conclusion The sFasL/Fas pathway may be one of the mechanisms by which DNT cells kill pancreatic cancer cells.