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 |
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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) |
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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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