摘要
目的探讨miR-374b在增强细胞因子诱导杀细胞(CIK)抗肿瘤中的作用及机制。方法采用传统方法培养CIK,用流式细胞确定CIK细胞群且分选出CIK;流式分析Hep G2、PLC、H1299、A549的程序性坏死蛋白(PD)-L1表达;采用双荧光素酶系统检测miR-374b在PD-1上的结合,siRNA技术干扰降低miR-374b基因的表达;酶联免疫吸附(ELISA)检测CIK的γ干扰素(IFN-γ)表达;乳酸脱氢酶(LDH)释放法检测CIK体外杀伤肿瘤细胞的能力;过继回输检测CIK体内杀伤肿瘤细胞的能力。结果双荧光素酶系统显示miR-374b可以直接作用于PD-1基因上;Hep G2、PLC、H1299、A549肿瘤细胞系都有比较高的PD-L1表达;miR-374b抑制剂会降低CIK分泌IFNr的水平,抑制CIK在体内和体外对肿瘤细胞的杀伤能力;PD-1 siRNA会提高CIK分泌IFN-γ的水平,增强CIK在体内和体外对肿瘤细胞的杀伤能力。结论 miR-374b可以直接作用于PD-1基因的表达,抑制miR-374b可以增强CIK杀伤肿瘤能力,干扰PD-1表达可以提高CIK抑制肿瘤能力。
引文
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