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目的:增强人肝癌细胞的免疫原性以激活宿主免疫细胞识别杀伤相应的肝癌细胞。方法:以脂质体介导小鼠H-2Kb基因转染肝癌细胞株,并检测H-2Kb抗原表达情况,然后用H-2Kb基因转染后的肝癌细胞体外激活效应细胞杀伤活性,再进行裸鼠动物实验。结果:H-2Kb基因转染人肝癌细胞株HepG2后,Southem印迹杂交结果显示,H-2Kb基因整合于肿瘤细胞染色体中,RNA点杂交结果显示,H-2KbDNA已转录成mRNA。免疫组化及流式细胞仪检测显示,H-2Kb抗原已在肝癌细胞胞膜上表达。转染后肝癌细胞在体外能强烈诱发效应细胞毒性,这种细胞毒性现象在裸鼠实验中得到进一步验证。结论:异源MHG-I类基因H-2Kb转染肝癌细胞后可增强其免疫原性并激活免疫效应细胞杀伤活性。
Objective: To enhance the immunogenicity of human hepatocellular carcinoma cells to activate the host immune cells to identify and kill the corresponding hepatoma cells. Methods: The H-2Kb gene was transfected into hepatocellular carcinoma cell line with liposome and the expression of H-2Kb antigen was detected. Then, the H-2Kb gene was used to infect the hepatocarcinoma cells in vitro. Nude mice experiment. Results: The H-2Kb gene was transfected into human hepatocellular carcinoma cell line HepG2. Southem blot showed that the H-2Kb gene was integrated into the chromosome of tumor cells. RNA hybridization showed that H-2KbDNA was transcribed into mRNA. Immunohistochemistry and flow cytometry showed that H-2Kb antigen was expressed on the membrane of hepatocellular carcinoma cells. After transfection, hepatocellular carcinoma cells can strongly induce effector cytotoxicity in vitro. This cytotoxicity has been further verified in nude mice. CONCLUSION: H-2Kb transfected into HepG2 cells can enhance its immunogenicity and activate immune effector cell killing activity.