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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
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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.
作者 姚秉彝 周倩 罗成凤 仲伟国 刘平 马成斌 YAO Bing-yi1,3, ZHOU Qian2, LUO Cheng-feng2, ZHONG Wei-guo3, LIU Ping3, MA Cheng-bin3 (1 Tongji University School of Medicine, Shanghai, 200092, China; 2 Shanghai First Maternity and Infant Hospital, Shanghai, 201204, China; 3 Shanghai Changning Maternity&Infant Health Hospital, Shanghai, 200051, China)
出处 《现代生物医学进展》 CAS 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)
关键词 肿瘤干细胞 卵巢癌 ALDH1 免疫抑制 肿瘤免疫 SKOV3 Cancer stem cells Ovarian cancer ALDH1 Immunosuppression Tumor immunity SKOV3
作者简介 姚秉彝(1981-),女,学士,主治医师,主要研究方向:妇科肿瘤,电话:13661903713,E-mail:13816852233@163.com 通讯作者:周倩,电话:13301762658,E—mail:shzhouqian@126.com
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