探究PHA-L共诱导的CIK细胞的生物学特性及其联合苦参碱体外抗胃癌效应

Biological characteristics of PHA-L-induced CIK cells and its anti-gastric cancer efficacy in combination with matrine in vitro

目的探讨植物血凝素-L(PHA-L)共诱导制备细胞因子活化的杀伤细胞(CIK)的方法,同时探究新型PHA-CIK细胞联合苦参碱体外对胃癌MGC-803细胞的杀瘤活性效应。方法通过PHA-L共诱导制备新型CIK细胞即PHA-CIK细胞,并比较分析新型CIK细胞与普通CIK细胞的生物学特性;同时采用CCK-8法分别检测普通CIK细胞组、PHA-CIK细胞组、苦参碱组以及PHA-CIK细胞联合苦参碱组(联合组)体外对胃癌MGC-803细胞的杀瘤活性。结果制备新型CIK的方法更能促进细胞增殖(P<0.05),且细胞活率保持在90%以上。与普通方法相比,新方法在CD3^+CD8^+比例方面存在显著差异(P<0.05),且杀伤活性增强(P<0.05)。联合组对胃癌MGC-803细胞的杀伤率明显高于PHA-CIK组和苦参碱组,分别是1.6倍和2.3倍,联合组与PHA-CIK组及苦参碱组相比差异具有显著统计学意义(P<0.01)。结论新型PHA-CIK联合苦参碱可显著提高对胃癌细胞的杀伤活性,为中药联合过继性细胞免疫治疗综合治疗肿瘤提供一种新方法。

This study aimed to explore a method to preparing cytokine-induced killer cells （CIK） by phytohemagglutinin-L （PHA-L）, and to investigate the anti-tumor activity of new PHA-CIK cells combined with matrine against human gastric cancer line MGC-803. The new PHA-CIK cells were induced by PHA-L. The biological characteristics of PHA-CIK cells and CIK cells were analyzed. CCK-8 was used to detect the anti-tumor activity of the CIK cells, PHA-CIK cells, matrine and PHA-CIK cells combined with matrine （combined group） in vitro. Compared with the normal method, the new method could promote cell proliferation （P〈0.05）, the prepared cell viability was kept above 90%, and CD3＋CD8＋ cells was increased （P〈0.05）. In combined group the rate of killing MGC-803 cells was 1.6 times and 2.3 times higher than that of PHA-CIK group and matrine group, respectively （P〈 0.01）. Taken together, the new PHA-CIK combined with matrine has significantly improved anti-tumor activity against gastric cancer cells, and provide a new treatment method for traditional Chinese medicine combined with adoptive cellular immunotherapy.