靶向PSCA的CAR-NK-92细胞对宫颈癌的抗瘤作用

Antitumor activity of chimeric antigen receptor NK-92 cells targeting PSCA anainst cervical cancer

目的:构建并验证靶向前列腺干细胞抗原(prostate stem cell antigen,PSCA)的嵌合抗原受体(chimeric antigen receptor,CAR)修饰的NK-92细胞对宫颈癌的抗瘤活性。方法:构建PSCA靶向的CAR慢病毒表达载体,利用慢病毒感染法获得PSCA CAR-NK-92细胞。用流式细胞术及Western blotting实验检测宫颈癌细胞中PSCA的表达水平。通过体外效靶细胞共孵育实验以及体内裸鼠移植瘤模型中的治疗情况,验证PSCA CAR-NK-92细胞对宫颈癌Hela及MS751细胞杀伤能力及其裸鼠移植瘤生长的抑制能力。结果:成功构建PSCA CAR-NK-92细胞,PSCA在宫颈癌细胞中高表达(均P<0.01)。体外共孵育结果显示,PSCA CAR-NK-92细胞可以以剂量依赖的方式裂解PSCA+的宫颈癌细胞;体内抗肿瘤数据表明,PSCA CAR-NK-92细胞较转染空载体的NK-92细胞显著抑制宫颈癌移植瘤的生长(P<0.01),并且可以有效地浸润肿瘤组织并高水平分泌TNF-α和IFN-γ(均P<0.01)。结论:靶向PSCA的CAR-NK-92细胞在体内外都显示出了良好的对PSCA+肿瘤细胞的杀伤能力,有潜力成为一种针对宫颈癌的潜在治疗策略。

Objective:To construct and verify the anti-tumor activity of chimeric antigen receptor(CAR)modified NK-92 cells(CAR-NK-92 cells)targeting prostate stem cell antigen(PSCA)in cervical cancer.Methods:Lentiviral vector expressing CAR targeting PSCA was constructed,and PSCA CAR-NK-92 cells were obtained by lentivirus transfection.The expression of PSCA in human cervical cancer cells was determined by Flow cytometry and Western blotting.The killing effect of PSCA CAR-NK-92 cells against cervical cancer cells was verified by co-incubation of effector and target cells in vitro,and the tumor inhibitory ability of PSCA CAR-NK-92 cells was verified with the nude mice xenograft model in vivo.Results:PSCA CAR-NK-92 cells were successfully constructed.PSCA was highly expressed in human cervical cancer Hela and MS751 cells(all P<0.01).In vitro co-incubation results showed that PSCA CAR-NK-92 cells could lyse PSCA+cervical cancer transplanted tumor in a dose-dependent manner.In vivo antitumor data showed that PSCA CAR-NK-92 cells significantly inhibited the growth of cervical cancer cells compared with NK-92 cells transfected with vehicle vectors(P<0.01).In addition,PSCA CAR-NK-92 cells could effectively infiltrate tumor tissues and promote the secretion of anti-tumor cytokines TNF-αand IFN-γ(all P<0.01).Conclusion:The CAR-NK-92 targeting PSCA shows good antitumor effect on PSCA+tumor cells both in vitro and in vivo,and has potential to be a therapeutic strategy for cervical cancer.