树突状细胞共培养因子诱导的杀伤细胞联合化疗治疗中晚期非小细胞肺癌的临床疗效

Clinical Efficacy of Dendritic Cells Co-cultured with Cytokine Induced Killer Cells Combined with Chemotherapy for Middle and Advanced Non-small Cell Lung Cancer

目的探讨树突状细胞与细胞因子诱导的杀伤细胞共培养对细胞因子诱导的杀伤细胞的作用及两者共培养后联合化疗药物奥沙利铂对中晚期非小细胞肺癌的体外杀伤效果。方法取非小细胞癌患者的恶性胸腔积液并采取密度梯度离心法获得树突状细胞,体外诱导获得成熟的树突状细胞;分离健康人的单核细胞,体外培养获取杀伤细胞;两者共培养后流式细胞鉴定表型,采用MTT法检测中晚期非小细胞肺癌的体外杀伤作用。结果恶性胸腔积液内树突前体细胞经过体外培养之后获得了成熟的树突细胞,培养9天以后获得树突细胞表明标志物CD83、CD80、CD86和MHC-II相关分子HLA-DR比培养前显著增高(P<0.05);培养2周之后树突细胞和杀伤细胞共培养与杀伤细胞单独培养比较CD3+、CD4+、CD8+数量明显增加,组间比较具有统计学差异(P<0.05)。树突细胞和杀伤细胞对中晚期非小细胞肺癌有着抑制作用,杀伤力大于单纯使用杀伤细胞组,组间比较具有统计学差异(P<0.05)。树突细胞和杀伤细胞共培养联合化疗药物奥沙利铂对中晚期非小细胞肺癌的体外杀伤具有联合增效的作用,组间比较具有统计学差异(P<0.05)。结论恶性胸腔积液来源的树突前体细胞可以诱导培养获得成熟的树突细胞,成熟的树突细胞联合杀伤细胞可以促进杀伤细胞的增殖及杀伤力,两者共培养联合化疗药物奥沙利铂具有协同增强中晚期非小细胞肺癌的体外杀伤作用。

Objective To investigate the effect of co-culture of dendritic cells and cytokine-induced killer cells on cytokine-induced killer cells,and the 2 co-cultured cells combined with oxaliplatin for advanced non-small cell lung cancer. Methods Patients with non-small cell lung cancer and malignant pleural effusions take density gradient centrifugation dendritic cells in vitro to obtain mature dendritic cells; from healthy patients monocytes in vitro obtained killer cells; 2 cells were identified by flow cytometry after co-culture phenotypes,in vitro cytotoxicity was analyzed by MTT assay in advanced non-small cell lung cancer. Results Malignant pleural effusion dendritic precursor cells were obtained after in vitro maturation of dendritic cells,dendritic cells cultured 9 days and showed markers CD83,CD80,CD86 and MHC-II related HLA-DR molecules than former culture were significantly higher( P < 0. 05); after 2 weeks in dendritic cells co-cultured with killer cells and killer cells cultured alone,CD3 +,CD4 +,CD8 + number increased significantly,there had statistically significant difference( P < 0. 05) between groups. Dendritic cells-killer cells in patients with advanced non-small cell lung cancer had inhibited the use of lethal than simply killer cells,there had statistical difference( P < 0. 05). Dendritic cells co-cultured with killer cells in combination with oxaliplatin for advanced non-small cell lung cancer has the effect of killing in vitro,there had statistically difference( P < 0. 05). Conclusion Malignant pleural effusion derived dendritic cell precursors can induce mature dendritic cells cultured,mature dendritic cells combined with killer cells can promote the proliferation and lethal of killer cells,combined with oxaliplatin can enhancement killing effect of advanced non-small cell lung cancer.