抗癌中药复方协同顺铂对Fas/FasL介导SKOV3细胞免疫逃逸的逆转作用

Compound herbal medicine prescription and cisplatin reversed Fas/FasL-mediated SKOV3 cell immunogic escape

目的研究抗癌中药复方(CHMP)药血清及其协同顺铂(DDP)对Fas/FasL介导SKOV3细胞免疫逃逸的逆转作用。方法根据前期研究结论制备抗癌中药复方(CHMP)1及CHMP2复方颗粒药最佳时相及剂量的药血清;观察CHMP与顺铂之间的协同作用;流式细胞术(FCM)分析对照组(A组)、CHMP1组(B组)、CHMP2组(C组)、顺铂组(D组)、CHMP1+顺铂(E组)、CHMP2+顺铂组(F组)等各组药血清对各实验组细胞SKOV3表面Fas/FasL分子及各实验组SKOV3细胞对JurkatT细胞凋亡的诱导。结果各组药血清处理72h后,SKOV3细胞表面Fas抗原表达率较对照组明显升高(P<0.01),SKOV3细胞表面FasL抗原表达率较对照组也显著降低(P<0.01)。但顺铂处理的SKOV3细胞FasL抗原表达与对照组相比显著提高(P<0.05)。SKOV3与JurkatT细胞共同培养时各组细胞凋亡率与对照组相比明显降低(P<0.05),联合用药与顺铂相比可减少JurkatT细胞的凋亡。结论 SKOV3细胞表面的FasL可能与JurkatT细胞表面的Fas结合,通过Fas/FasL途径诱导T淋巴细胞凋亡来逃避免疫系统的攻击。联合用药与顺铂相比可减少JurkatT细胞的凋亡,从而增加其对肿瘤细胞的杀伤活性。与化疗药顺铂具有协同作用,解毒化瘀复方与顺铂的协同作用优于补益复方。

Objective To investigate the effects of serum containing anti-tumorigenic compound herbal medicine prescription（CHMP） in combination with cisplatin（DDP） on reversing Fas/FasL-mediated immunologic escape in human ovarian carcinoma cell line（SKOV3）.Methods Serum was obtained at the optimal time and dosage of tonifying herbal granule preparation（CHMP1） and detoxicating and blood circulation activating herbal granule preparation（CHMP2） based on the literatures previously,for determination of the synergetic effect of CHMP and DDP.Flow cytometry was employed to analyze the effects of serum in control group（group A）,CHMP1 group（group B）,CHMP2 group（group C）,DDP group（group D）,CHMP1＋DDP group（group E） and CHMP2＋DDP group（group F） on SKOV3 surface Fas/FasL and subsequent induction of Jurkat T cell apoptosis.Results Higher Fas and FasL antigen expression on SKOV3 cell surface was noted at hour 72 after treatment in all groups with exception of group A（all P〈0.01）.DDP-treated SKOV3 cells were found as having considerably higher expression rate of FasL antigen than those in group A（P〈0.05）.Coculturing of SKOV3 and Jurkat T cells yielded the apoptotic rates of（34.2±2.2）%,（10.2±1.4）%,（13.0±1.7）%,（21.1±3.2）%,（15.2±3.7）% and（14.2±4.2）% in group A,B,C,D,E and F,respectively.In contrast,group A was associated with significantly lower apoptotic rate（all P〈0.05）.Combination therapy,but not DDP alone,resulted in minor Jurkat T cell apoptosis.Conclusion SKOV3 surface FasL may escape immunologic attack by integrating with Jurkat T surface Fas and subsequent induction of T lymphocyte apoptosis via the Fas/FasL pathway.Compared with DDP,combination therapy achieves minor Jurkat T cell apoptosis and higher anti-tumorigenic activity and could have synergistic effect with DDP combined.Additionally,superior synergetic effect may be associated with detoxicating and blood circulation activating,but not toniying preparation.