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组蛋白去乙酰化酶抑制剂在胰腺癌细胞免疫治疗中的意义

, PP. 2389-2394

Keywords: 组蛋白去乙酰化酶(HDAC)抑制剂,胰腺癌,生物治疗,CIK,MICA/B

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Abstract:

目的研究组蛋白去乙酰化酶(histone-deacetylase,HDAC)抑制剂增强细胞免疫疗法治疗胰腺癌的机制和意义。方法将PANC-1、BXPC-3、CFPAC-1胰腺癌细胞株分别设定不加药物组,HDAC抑制剂[曲古抑菌素A(TSA)]加药组。使用流式细胞计数、免疫荧光染色计数、Westernblot分别测定两组胰腺癌细胞株的模型MICA/B表达。测定不同浓度、不同作用时间TSA对3种胰腺癌细胞株MICA/B表达的影响。找到TSA调高胰腺癌细胞株MICA/B表达的最佳浓度为200nmol/L,最佳作用时间为24h。乳酸脱氢酶释放计量法测定CIK细胞∶肿瘤细胞=10∶1的情况下CIK细胞对两组胰腺癌细胞株的杀伤率。双变量相关性分析CIK细胞对胰腺癌细胞株杀伤率与胰腺癌细胞株MICA/B的表达的关系。结果流式细胞仪检测的PANC-1、BXPC-3、CFPAC-1的MICA/B表达率分别为2.2%、61.2%、31.2%。200nmol/LTSA处理24h后,表达率分别为13.8%、86.6%、76.1%(P<0.01)。免疫荧光染色显示TSA处理后的细胞染色明显增强;Westernblot检测TSA处理后的胰腺癌细胞株MICA/B膜型蛋白表达增强。在CIK细胞∶肿瘤细胞=10∶1的情况下,CIK细胞对PANC-1、BXPC-3、CFPAC-1的杀伤率分别为7.66%、12.03%、12.15%。200nmol/LTSA处理24h后,CIK对以上3种细胞的杀伤率为24.32%、58.63%、49.25%(P<0.01)。CIK细胞对PANC-1、BXPC-3、CFPAC-1细胞的杀伤率与细胞表面膜型MICA/B的表达的相关性系数分别为0.959、0.964、0.972(P<0.01)。表明与CIK细胞对胰腺癌细胞株的杀伤率与胰腺癌细胞株的细胞表面膜型MICA/B的表达呈正相关。结论HDAC抑制剂通过调高胰腺癌细胞株的MICA/B表达来增强CIK细胞对胰腺癌细胞株的杀伤,从而提高胰腺癌细胞免疫治疗的效果。

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