Sonic hedgehog阻断抗体增强外周血单个核细胞抗宫颈癌HeLa细胞的作用

Anti-Sonic hedgehog blocking antibody enhances killing effect of PBMCs on cervical carcinoma HeLa cells

目的:探讨Sonic hedgehog(Shh)阻断抗体对外周血单个核细胞(PBMCs)抗宫颈癌HeLa细胞作用的影响。方法:免疫细胞化学技术和RT-PCR法检测Shh及其信号分子在HeLa细胞中的表达;Ficoll密度梯度离心法分离正常人PBMCs,并与HeLa细胞建立共培养体系;于共培养体系中加入Shh阻断抗体,流式细胞术检测CD3、CD69和CD71分子表达;ELISA法检测细胞因子IFN-γ、IL-10和IL-4分泌;显微镜观察PBMCs对HeLa细胞的杀伤。结果:HeLa细胞表达Shh及其信号分子,HeLa细胞传代6次对Shh的表达水平没有影响;共培养体系中CD3+细胞增多;Shh阻断抗体可以促进CD69、CD71表达和IFN-γ的分泌,但抑制IL-10分泌,对IL-4分泌没有显著影响;Shh阻断抗体增强PBMCs对HeLa细胞的杀伤。结论:Shh阻断抗体可以促进PBMCs活化,增强PBMCs抗宫颈癌HeLa细胞的作用。

AIM： To investigate the effect of anti-Sonic hedgehog（Shh） blocking antibody on the killing effect of peripheral blood mononuclear cells（PBMCs） on cervical carcinoma HeLa cells.METHODS： The expression levels of Shh and Shh signaling molecules in HeLa cells were detected by immunocytochemistry and RT-PCR.PBMCs from health peoples were isolated by the method of Ficoll density gradient centrifugation,and then co-cultured with HeLa cells in vitro.The expression of CD3,CD69 and CD71 was assayed by flew cytometry.The concentrations of IFN-γ,IL-10 and IL-4 in culture supernatants were detected by ELISA.The killing effect of PBMCs on HeLa cells was observed under microscope.RESULTS： Shh and Shh signaling molecules were expressed in HeLa cells.The level of Shh expression didn＇t change significantly in the 6th passage of HeLa cells.CD3＋ cells were increased in the co-culture system.The expression of CD69 and CD 71,and the secretion of IFN-γ were increased,while the secretion of IL-10 was decreased in the co-culture system treated with anti-Shh blocking antibody.Anti-Shh blocking antibody has no effect on the secretion of IL-4.The killing effect of PBMCs on HeLa cells was strengthened by anti-Shh blocking antibody.CONCLUSION： Anti-Shh blocking antibody promotes the activation of PBMCs and enhances the killing effect of PBMCs on cervical carcinoma HeLa cells.