卵巢癌患者肿瘤浸润淋巴细胞的抗肿瘤活性观察

发布时间：2019-08-15 17:11

【摘要】：目的观察卵巢癌(OC)患者肿瘤浸润淋巴细胞(TILs)的抗肿瘤活性。方法采用联合酶消化分离并提纯OC组织中的TILs和OC细胞,以IL-2体外诱导活化扩增TILs。活化前及活化第14天,采用流式细胞仪检测TILs免疫表型(CD+3、CD+4、CD+8、CD+4/CD+8);活化前及活化第3、7、14天,采用MTT法检测TILs对自体OC细胞、人白血病细胞株K562、人B细胞淋巴瘤细胞株Raji的杀伤活力;收集经活化第14天TILs作用前后的OC细胞培养上清液,ELISA法检测肿瘤分子标志物CA-125、CA-153、血管内皮生长因子(VEGF)、基质金属蛋白酶9(MMP-9)。结果与活化前比较,活化第14天TILs中CD+3、CD+4、CD+8细胞比例增加,CD+4/CD+8细胞比值降低;随着活化时间延长,TILs对三种靶细胞的杀伤率增高(P均0.05),且对OC细胞的杀伤率高于其余2种细胞(P均0.05);与处理前相比较,活化第14天TILs作用的OC细胞培养上清液中CA-125、VEGF、MMP-9水平均显著降低(P均0.05)。结论卵巢癌患者的TILs具有抗肿瘤活性,低浓度IL-2可增强其特异性和非特异性抗瘤活性。
[Abstract]:Objective to observe the antitumor activity of tumor infiltrating lymphocytes (TILs) in patients with ovarian cancer (OC). Methods TILs and OC cells in OC tissue were digested and purified by combined enzyme digestion and purification. TILs. was activated and amplified by IL-2 in vitro. The immunophenotypes of TILs (CD 3, CD 4, CD 8, CD 4/CD 8) were detected by flow cytometry before activation and 14 days after activation, and the killing activity of TILs against autologous OC cells, human leukemia cell line K562 and human B cell lymphoma cell line Raji was detected by MTT assay before activation and on the 3rd, 7th and 14th day of activation. The culture media of OC cells before and after activation of TILs on the 14th day were collected and the tumor molecular marker CA-125,CA-153, vascular endothelial growth factor (VEGF), matrix metalloprotease 9 (MMP-9) was detected by ELISA assay. Results on the 14th day after activation, the proportion of CD 3, CD 4, CD 8 cells and the ratio of CD 4/CD 8 cells in TILs increased, and the killing rate of TILs to three kinds of target cells increased with the prolongation of activation time, and the killing rate to OC cells was higher than that of the other two kinds of cells. Compared with those before treatment, the level of CA-125,VEGF,MMP-9 in the culture medium of OC cells treated with TILs on the 14th day was significantly decreased (P < 0.05). Conclusion TILs has antitumor activity in patients with ovarian cancer. Low concentration of IL-2 can enhance its specific and nonspecific antitumor activity.
【作者单位】：南方医科大学南方医院;南方医科大学第三附属医院;
【基金】：广东省科技计划资助项目(2011B031800042)
【分类号】：R737.31

【参考文献】