卵巢上皮性癌细胞系来源肿瘤干细胞的免疫特性研究  

Research on the Immune Response of Ovarian Cancer Cell Line Derived Cancer Stem Cells

作　　者：姚秉彝 周倩[3] 罗成凤[3] 仲伟国 刘平 马成斌 

机构地区：[1]同济大学医学院,上海200092 [2]上海市长宁区妇幼保健院,上海200051 [3]同济大学附属上海市第一妇婴保健院,上海201204

出　　处：《现代生物医学进展》2017年第2期205-209,共5页Progress in Modern Biomedicine

基　　金：上海市自然科学基金项目(16ZR1427300)

摘　　要：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： The dimensional cultures（spheroids） were generated from Ovarian cancer cell lines（SKOV3） in order to enrich for CSCs. Methods： Stem cell markers ALDH1 were compared between CSC-enriched spheroid culture-derived cells（SDC） and the corresponding monolayer culture-derived cells（MDC）. We evaluated the effect on proliferation and function of SDC or MDC on T cells in transwell assays. To compared the immune suppression gene expression of the SDC and MDC in RT-PCR assays. Results： We show that CSC-enriched SDC populations exhibit a higher proportion of ALDH1 expressing cells as compared with corresponding MDCs. The proliferation rate of T cells cocultured with SDC was significantly less than that of T cells cocultured with MDC. The expression of cytokine production of IFN-γ and IL-2 by T cells cocultured with SDC was significantly less than that of T cells cocultured with MDC（P0.05）. RT-PCR results showed that the expression of SDC gene, such as ArginaseⅡ, IDO, IL-8 and TGF-β,was significantly higher than that of MDC,specially in ArginaseⅡ and IL-8. Conclusion： Spheroid culture is a reliable method for obtaining ovarian tumor stem cells. Consistently, we have found in ovarian cell lines（SKOV3） a strong immunosuppressive activity exerted by CSC. All T cell functions investigated were suppressed upon co-culture with SDC significantly stronger than by MDC. This maybe an t escape mechanism from immune cells supporting sustained growth of tumors and hampering immune therapeutic strategies.

关 键 词：肿瘤干细胞 卵巢癌 ALDH1 免疫抑制 肿瘤免疫 SKOV3 

分 类 号：R33[医药卫生—人体生理学] R737.31[医药卫生—基础医学]