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目的通过将趋化因子Fractalkine(FK)基因转染小鼠肝癌细胞株MM45T.Li,探讨FK用于诱导抗肝癌主动免疫的可行性。方法用脂质体将小鼠FK基因导入小鼠肝癌细胞株MM45T.Li,G418筛选抗性克隆。逆转录聚合酶链反应检测FK的表达,体内实验观察野生型及FK基因修饰肿瘤细胞的致瘤性, 组织病理学检测肿瘤中免疫细胞的浸润,流式细胞仪检测外周血中CD4+、CD8+ T淋巴细胞水平。结果逆转录聚合酶链反应检测发现G418筛选得到的转染FK的抗性克隆MM45T.Li-FK表达FK,而野生型MM45T.Li不表达FK。体内成瘤实验显示,与野生型瘤细胞相比,MM45T.Li-FK的成瘤率显著下降。MM45T.Li-FK所形成肿瘤中有明显的免疫细胞浸润,对照组肿瘤中免疫细胞的浸润较少。接种MM45T.Li- FK瘤细胞的小鼠,外周血中CD4+、CD8+ T淋巴细胞水平明显增高,与对照组相比差异均有统计学意义(P <0.01)。结论转染FK基因能促进机体的抗肝癌主动免疫反应。
Objective To investigate the feasibility of using FK to induce active immunity against hepatocellular carcinoma (HCC) by transfecting mouse hepatoma cell line MM45T.Li with the gene of fractalkine (FK). Methods Mouse FK gene was transfected into mouse hepatoma cell line MM45T.Li by lipofectamine. The resistant clone was screened by G418. The expression of FK was detected by RT-PCR. The tumorigenicity of wild-type and FK-modified tumor cells was observed in vivo. The infiltration of immune cells in tumor was detected by histopathology. The levels of CD4 +, CD8 + T Lymphocyte levels. Results Reverse transcriptase-polymerase chain reaction assay showed that the F4-resistant F45-resistant clone clone MM45T.Li-FK expressed FK, whereas wild-type MM45T.Li did not express FK. In vivo tumorigenicity assays showed a significant decrease in the tumorigenicity of MM45T.Li-FK compared to wild-type tumor cells. The tumors formed by MM45T.Li-FK had obvious immune cell infiltration, while the immune cells in the control group had less infiltration. The mice inoculated with MM45T.Li-FK cells had significantly increased levels of CD4 + and CD8 + T lymphocytes in peripheral blood, which were significantly different from those in control group (P <0.01). Conclusion Transfection of FK gene can promote the body’s active immune response against liver cancer.