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目的:探讨卵巢上皮性癌细胞系(SKOV3)来源肿瘤干细胞的免疫特性。方法:通过干细胞特异性标记物乙醛脱氢酶1ALDH1表达鉴定富集肿瘤干细胞(CSCs)的球状细胞(SDC)。将富集CSCs的SDC与相应的贴壁培养的普通卵巢癌细胞(MDC)进行比较。通过Transwell法(细胞侵袭试验)检测SDC和MDC对于T细胞增殖以及分泌功能的影响。RT-PCR检测SDC以及MDC免疫抑制基因表达情况。结果:以球体细胞形成方法获得的CSC富集SDC群比MDC表现出更高比例的ALDH1表达。将T细胞分别与SDC和MDC共培养,SDC组T细胞增殖率显著低于MDC组。MDC共培养的T细胞分泌IFN-γ、IL-2水平明显高于与SDC共培养的T细胞(P<0.05)。SDC表达免疫抑制基因ArginaseⅡ、IDO、IL-8、TGF-β水平明显高于MDC,以ArginaseⅡ、IL-8的上调最为明显(P<0.05)。结论:球体细胞形成试验是获得卵巢肿瘤干细胞的可靠方法。在卵巢上皮性癌细胞系(SKOV3)中,CSCs比普通肿瘤细胞表现出更强大的免疫抑制性,这可能是妨碍免疫治疗的免疫逃逸机制。
Objective: To investigate the immunological characteristics of cancer stem cells derived from ovarian epithelial carcinoma cell line (SKOV3). METHODS: Splenocytes (SDCs) enriched in tumor stem cells (CSCs) were identified by 1ALDH1 expression of the stem cell specific marker aldehyde dehydrogenase. The CSCs-enriched SDCs were compared with the corresponding adherent cultured normal ovarian cancer cells (MDCs). The effects of SDC and MDC on T cell proliferation and secretion were tested by Transwell method (cell invasion assay). RT-PCR detection of SDC and MDC immunosuppressive gene expression. Results: The CSC-enriched SDC population obtained by the somatic cell formation method showed a higher proportion of ALDH1 expression than MDC. T cells were co-cultured with SDC and MDC respectively. The proliferation rate of T cells in SDC group was significantly lower than that in MDC group. The levels of IFN-γ and IL-2 secreted by MDC co-cultured T cells were significantly higher than those co-cultured with SDC (P <0.05). The expression of ArginaseⅡ, IDO, IL-8 and TGF-β in SDC was significantly higher than that in MDC, and the upregulation of ArginaseⅡ and IL-8 was the most significant (P <0.05). CONCLUSIONS: Spheroblast formation assays are a reliable method of obtaining ovarian cancer stem cells. In ovarian epithelial carcinoma cell line (SKOV3), CSCs showed more potent immunosuppression than normal tumor cells, which may be an immune escape mechanism that hinders the immunotherapy.