摘要
目的 :用肿瘤坏死因子 (Tum or necrosis factor- α,TNF- α)基因转染人肝癌肿瘤浸润淋巴细胞 (Tumor infiltratinglym phocyte,TIL) ,构建转基因 TIL,对其生物学特性进行研究 ,为肝癌基因治疗奠定基础。方法 :用逆转录病毒载体将 TNF- α基因导入人肝癌肿瘤浸润淋巴细胞 ,构建一种新型的抗肿瘤效应细胞 -转基因 TIL。对转基因 TIL 的增殖能力和细胞表型进行检测 ,用 RT- PCR检测目的基因的表达 ,TNF- α试剂盒检测 TNF- α分泌量 ,用 MTT法检测转基因 TIL 体外抗肿瘤活性。结果 :转基因 TIL 的增殖活性及细胞表型与 IL- 2活化的 TIL 相似 ,转基因 TIL 可持续表达 TNF- α,其分泌量每 2 4 h达 370 pg/5× 10 5cells。转基因 TIL 对肝癌细胞株 BEL74 0 4具有较高的杀伤活性。结论 :转基因 TIL 维持较高的稳定性 ,可持续表达TNF- α,具有良好的体外抗肿瘤作用 。
Objective: Transduced TIL was constructed based on tumor infiltrating lymphocytes (TIL) transduced with tumor necrosis factor α (TNF α) gene The biological characters of transduced TIL was studied, and the base of gene therapy in hepatocellular carcinoma was established Methods: To construct transduced TIL, retroviral vector was used to introduce TNF α into TIL which was isolated from human hepatocellular carcinoma tissue The proliferation activity and the cell phenotype of transduced TIL was detect The expression of aim gene was detected by RT PCR, and TNF α detecting kit was utilized to detected the secretion of TNF α The cytotoxic activity of transduced TIL was detected by MTT method Results: The proliferation activity and cell phenotype of transduced TIL was similar to TIL which activated by IL 2 Transduced TIL could secret TNF α continuously, its expression dose was 370 pg/5×10\+5 cells/24 h Transduced TIL had higher cytotoxic activity against human hepatoma cell line BEL\-\{7404\} Conclusion: Transduced TIL maintain highly stability, and can secret TNF α continuously The anti tumor cytotoxic activity of transduced TIL was better in vitro, and the application for treatment of hepatocelluar carcinoma has a good future
出处
《广西医科大学学报》
CAS
2002年第2期151-154,共4页
Journal of Guangxi Medical University
基金
广西教育厅重点科研资助项目 (19982 -8)
关键词
生物学
肿瘤浸润淋巴细胞
肿瘤坏死因子
肝癌
基因治疗
tumor infiltrating lymphocyte
tumor necrosis factor
gene therapy
hepatocellular carcinoma
