IDO基因转染宫颈癌细胞SiHa在体外抑制NK细胞杀伤作用的研究

摘要

目的 通过IDO基因转染,研究IDO在体外对NK细胞杀伤能力的抑制作用.方法 IDO蛋白表达和空白质粒分别稳定转染至人宫颈癌细胞SiHa,应用Western blot检测IDO在SiHa、SiHa/Mock和SiHa/IDO细胞中的表达情况,用MTT试剂盒检测3组肿瘤细胞体外生长速度和对NK细胞杀伤作用的敏感性.结果 IDO蛋白在SiHa/IDO细胞中表达阳性,在SiHa和NHa/Mock细胞中无表达.3组肿瘤细胞体外生长曲线无统计学差异,P＞0.05.SiHa/IDO细胞存活的百分比高于其他2组对照(SiHa和SiHa/Mock)细胞,P＜0.05.结论 DO对宫颈癌细胞体外生长速度无影响,但可抑制宫颈癌细胞SiHa对NK细胞杀伤作用的敏感性.因此,IDO不仅可以作为宫颈癌判断预后的指标,同时也可以作为宫颈癌基因治疗的潜在新靶点.%Objective To investigate the effect of Indoleamine 2,3-dioxygenase (IDO J expression on the sensitivity of cervical cancer cells against natural killer ( NK) cells after IDO gene transfer. Methods SiHa cells were stably transfect-ed with IDO expression vector and Mock vector. Western blot analysis was used to detect IDO expression in SiHa, SiHa/ Mock and SiHa/IDO cells. Cell growth curves were performed using MTT kit. Sensitivity of SiHa, SiHa/Mock and SiHa/ IDO cells to NK cells were investigated by colorimetric assay using MTT. Results Only SiHa/IDO could express IDO. There was no intergroup difference in doubling time in vitro (P >0.05). The percent survival of SiHa/IDO cells was significantly higher than those of the control (SiHa and SiHa/Mock) cells (P<0.05). Conclusions IDO expression did not influence the cell growth of SiHa cells in vitro. IDO expression inhibited the cytotoxic activity of NK cells against cervical cancer cells. Therefore, IDO acts not only as a prognostic factor of cervical cancer but also as a potential therapeutic gene target against cervical cancer.