PSMA-CAR-CIK转基因细胞体外杀伤三种前列腺癌细胞株效率的研究

Efficacy of PSMA-CAR-CIK transgenic cells in killing three prostate cancer cell lines in vitro

目的:本研究通过检测前列腺特异性膜抗原(prostate specific membrane antigen,PSMA)-肿瘤特异性嵌合抗原受体(chimeric antigen receptor,CAR)-细胞因子诱导的杀伤细胞(cytokine induced killer cell,CIK)转基因细胞杀伤三种前列腺癌细胞株的效率,鉴定其用于后期治疗前列腺癌的可行性。方法:通过已经构建的二代PSMA-CAR慢病毒表达载体转染CIK细胞,构建PSMA-CAR-CIK转基因细胞;通过CCK8法检测CIK细胞及PSMA-CAR-CIK转基因细胞杀伤三种前列腺癌细胞株PC3、LNCaP、DU145的效率及效靶比。结果:转基因PSMA-CAR-CIK细胞构建成功;3 h~3.5 h为检测效应细胞对靶细胞细胞毒活性时与CCK8试剂孵育的最佳时间;两种效应细胞均在效靶比为10∶1、15∶1、20∶1时,杀伤率逐步增高,无统计学意义(P>0.05),但与其它组相比差异显著(P<0.05)。结论:两种效应细胞对于前列腺癌细胞的杀伤,只要效靶比达到10∶1的比例,就能起到一个较好的杀瘤效果,并且随着效应细胞的增加,其抗肿瘤能力逐步加强;转基因PSMA-CAR-CIK细胞的构建为CAR技术治疗前列腺癌的临床应用奠定了基础。

Objective:To examine the killing efficiency of three prostate cancer cell lines by PSMA-CAR-CIK transgenic cells,and evaluate the feasibility of PSMA-CAR-CIK for later treatment of prostate cancer.Methods:CIK cells were transfected with the second-generation PSMA-CAR lentivirus expression vector,and PSMA-CAR-CIK transgenic cells were constructed.The efficiency and effect-target ratio of CIK cells and PSMA-CAR-CIK transgenic cells kill PC3,LNCaP and DU145 prostate cancer cells were detected by CCK8 assay.Results:The transgenic PSMA-CAR-CIK cells were successfully constructed.3 h~3.5 h was the best incubation time with CCK8 reagent for detecting the cytotoxic activity of effector cells against target cells.The killing rate of the two effector cells increased gradually at the effect-target ratio of 10∶1,15∶1 and 20∶1,with no statistical significance(P>0.05),but the difference was significant compared with other groups(P<0.05).Conclusion:As long as the ratio of effect to target reach 10∶1,the two kinds of effector cells could have a good anticancer effect on prostate cancer cells,and with the increase of effector cells,their anti-tumor ability is gradually enhance.The construction of transgenic PSMA-CAR-CIK cells lays the foundation for the clinical application of CAR technology in the treatment of prostate cancer.