文章摘要
姚秉彝 周倩 罗成凤 仲伟国 刘平 马成斌.卵巢上皮性癌细胞系来源肿瘤干细胞的免疫特性研究[J].,2017,17(2):205-209
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 DerivedCancer StemCells
  
DOI:
中文关键词: 肿瘤干细胞  卵巢癌  ALDH1  免疫抑制  肿瘤免疫  SKOV3
英文关键词: Cancer stemcells  Ovarian cancer  ALDH1  Immunosuppression  Tumor immunity  SKOV3
基金项目:上海市自然科学基金项目(16ZR1427300)
作者单位
姚秉彝 周倩 罗成凤 仲伟国 刘平 马成斌 同济大学医学院同济大学附属上海市第一妇婴保健院上海市长宁区妇幼保健院 
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中文摘要:
      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-酌、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-茁水平明显高于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。
英文摘要:
      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-r and IL-2 by T cells cocultured with SDC was significantly less than that of T cells cocultured with MDC (P<0.05). RT-PCR results showed that the expression of SDC gene, such as ArginaseII, IDO, IL-8 and TGF-beta,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 mechanismfromimmune cells supporting sustained growth of tumors and hampering immune therapeutic strategies.
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