CD4^+CD25^+FoxP3^+调节性T细胞在小鼠H22原位肝癌模型中的表达和意义

Expression and Its Significance of CD4^+CD25^+FoxP3^+T Regulatory Lymphocytes in H22 Orthotopic Tumor Model of Hepatocellular Carcinoma in Mice

目的检测小鼠原位肝癌模型中CD4+CD25+FoxP3+调节性T细胞的表达,探讨其在肝癌免疫中的作用。方法 Balb/C小鼠随机分为肝癌模型组、生理盐水对照组、空白对照组各10只,采用细胞悬液直接注射法制成原位肝癌模型。模型制作后第30天处死动物:流式细胞术检测分离纯化后的各组新鲜肝组织和脾脏CD4+CD25+T细胞的相对比例;实时RTPCR检测各组肝组织FoxP3基因表达水平。结果肝癌模型组肿瘤组织CD4+CD25+T/CD4+T细胞的比例增高,与肝癌模型组癌旁远端组织、生理盐水对照组肝组织、空白对照组肝组织比较,差异有统计学意义(P<0.05),而各对照组之间的比例差异无统计学意义(P>0.05)。肝癌模型组脾脏CD4+CD25+T/CD4+T细胞的比例增高,与生理盐水对照组脾脏、空白对照组脾脏比较,差异有统计学意义(P<0.01),而各对照组之间的比例差异无统计学意义(P>0.05)。肝癌模型组肿瘤组织FoxP3mRNA的表达水平高于肝癌模型组癌旁远端组织、生理盐水对照组肝脏组织和空白对照组肝脏组织,差异有统计学意义(P<0.01),而各对照组之间FoxP3mRNA的表达水平差异无统计学意义(P>0.05)。结论 Treg细胞在小鼠原位肝癌模型中表达升高,且对细胞免疫起抑制作用。

Objective To detect the expression of CD4 CD25 ＋ FoxP3 T regulatory lymphocytes in orthotopic tumor models of hep- atocellular carcinoma in mice, and investigate the role of it in hepatocellular carcinoma immunization. Methods Balb/C mice were divided into groups： tumor model of hepatocellular carcinoma group, normal sodium control group,blank group, with 10 in every group. Orthotopic tumor models of hepatoeellular carcinoma in mice were constructed by direct injection. Thirty days after the models, being made all mice were sacrificed. Fresh liver tissue and spleen tissue were separated and purified, Then CD4 ＋ CD25 ＋ T cells of the tissues were detected by using flow eytometry. FoxP3 genic expressoion of the liver tissue was detected by reahime - PCR. Results CD4 CD25 ＊ T/CD4 ＋T cells of the cancer tissues in the tumor models of hepatocellular carcinoma group increased. Compared with control group, the difference had statistical significance（ P 〈 0.05） , But differences in the relative proportions of the alls the liver tissue far from cancer, in the live tissue of normal sodium control group, and in the liver tissue of black group were not significant （P 〉 0.05）. Relative proportions of CD4 ＋ CD25 T/CD4 T cells in the spleen tissue in the tumor models of hepatoeellular carcinoma group rose up. Compared with control group, the differences had significant statistical significance（ P 〈 0.01 ） , But differences in the relative proportions between the spleen tissue of normal sodium control group and the spleen tissue of black group were not significant （ P 〉 0.05 ）. FoxP3 genic expression of the cancer tissues was higher than the other control group, and the differences had significant statistical significance （P 〈 0.01 ）. But FoxP3 genie expression differences among the liver tissues of black group, of normal sodium control group and the liver tissues around cancer were not significant （P 〉 0.05 ）. Conclusion The expression of Treg cells in orthotopic tumor models of hepatocellular carcinoma in mice increases, and has the function of suppressing the cellular immune.