HLA-G在卵巢癌组织中的表达及对NK细胞杀伤活性的影响  

HLA-G expression in human ovarian cancer and its roles in NK-92 cell cytotoxicity

作　　者：王斌梁[1] 章霞[1] 许惠惠[1] 颜卫华[1] 林爱芬[1] 

机构地区：[1]温州医学院附属浙江省台州医院心胸外科,临海317000

出　　处：《现代免疫学》2011年第6期456-461,共6页Current Immunology

基　　金：浙江省科技计划资助项目(2008C33013;2009C33147)

摘　　要：通过免疫组织化学法(IHC)检测卵巢癌组织中HLA-G的表达;克隆编码全长HLA-G1cDNA,将该基因通过真核表达载体pVITRO2-mcs转染至HLA-G阴性的HO-8910、OVCAR-3细胞株,采用RT-PCR、流式细胞术(FACS)及Westernblot鉴定、分析转染细胞中HLA-G的mRNA及蛋白质表达;LDH释放法检测卵巢癌细胞表达HLA-G后对NK细胞杀伤功能的影响。结果显示,66.7%(22/33)卵巢癌组织表达HLA-G分子,而正常组织未见其表达;基因转染后,HLA-G在HO-8910及OVCAR-3细胞表面获得稳定表达。细胞毒实验结果显示,HLA-G能显著抑制NK-92对卵巢癌细胞的杀伤活性(P<0.01);HLA-G特异性抗体87G阻断后,能明显恢复NK-92细胞对HO-8910-G、OVCAR-3-G的杀伤功能。本研究结果提示,卵巢癌细胞通过表达HLA-G分子抑制NK细胞的杀伤活性,在卵巢癌细胞逃避宿主的免疫监视中起重要作用。To analyze HLA-G expression in ovarian cancer tissues and to investigate its roles in NK-92 cell cytotoxicity with the ovarian cancer cell lines HO-8910 and OVCAR-3,HLA-G expression in ovarian cancer tissues was analyzed by immunohistochemistry.Full length HLA-G1 cDNA was cloned to the eukaryotic expression vector pVITRO2-mcs,and transfected to HLA-G negative HO-8910 and OVCAR-3 cells by Lipofectamine.HLA-G expression was confirmed by RT-PCR,flow cytometry and Western blot assay.NK cell cytotoxicity was performed with the LDH releasing method.It was found that HLA-G expression was detected in 22/33（66.7%） of primary tumor tissues,but it was absent in normal ovarian tissues.Cytotoxicity studies showed that HLA-G expression dramatically inhibited cell lysis by NK-92 cells（P0.01）,which could be restored by the anti-HLA-G conformational mAb 87G（P0.01）.HLA-G was expressed in a significant number of primary ovarian cancer tissues,and HLA-G expression in HO-8910 and OVCAR-3 could directly inhibit NK-92 cell cytotoxicity.Taken together,our results indicated that HLA-G expression plays an important role in the evasion of ovarian cancer cells from host immunosurveillance.

关 键 词：卵巢癌 HO-8910 OVCAR-3 HLA-G NK-92细胞 

分 类 号：Q343.1[生物学—遗传学]