人LIGHT和HSV-TK-EGFP基因共转染的MSCs抗肿瘤免疫的体内研究  

Anti-tumor Effect of Mesenchymal Stem Cells Modified by LIGHT and HSV-TK-EGFP Genes in Vivo

作　　者：陈芬[1,2] 江千秋[1,2] 焦兰[3] 唐澍[1] CHEN Fen;JIANG Qianqiu;JIAO Lan(Laboratory for Regenerative Medicine of Nanhua University,Chenzhou,423000)

机构地区：[1]南华大学转化医学研究所,423000 [2]湖南省郴州市第一人民医院,423000 [3]南华大学附属第一医院,421001

出　　处：《实用癌症杂志》2024年第11期1744-1748,共5页The Practical Journal of Cancer

摘　　要：目的研究与单纯疱疹病毒的糖蛋白D竞争结合单纯疱疹病毒进入介导物(herpes virus entry mediator,HVEM)的淋巴毒素类似物(homologous to lymphotoxins,exhibits inducible expression,and competes with HSV glycoprotein D for HVEM,a receptor expressed by T lymphocytes,LIGHT)基因和单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,HSV-TK)基因共转染的骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体内的抗肿瘤免疫功能。方法将pIRES2-LIGHT基因和HSV-TK-EGFP基因共转染小鼠骨髓间充质干细胞(MSCs/LT组),以转染空载体和转染HSV-TK-EGFP基因的骨髓间充质干细胞作对照。流式细胞仪检测LIGHT分子和HSV-TK-EGFP分子在稳定转染的骨髓间充质干细胞上的表达。体内迁移实验观察MSCs/LT在小鼠体内迁移情况。观察更昔洛韦注射前后MSCs/LT对荷瘤小鼠体内肿瘤的治疗作用。ELISA法检测小鼠肿瘤组织中IFN-γ,IL-2和IL-10的水平。结果流式细胞仪检测发现,MSCs/LT能稳定高表达LIGHT分子。MSCs/LT有特异地向肿瘤组织趋化的特性。MSCs/LT和MSCs/T有较好的抑制肿瘤生长的能力,但在更昔洛韦诱导后,MSCs/LT的抗肿瘤效应下降甚至消失。同时,MSCs/LT可促使T细胞进入肿瘤组织,并促进T细胞分泌IL-2、IFN-γ,抑制IL-10分泌(P<0.05)。结论共转染人LIGHT和HSV-TK-EGFP基因的骨髓间充质干细胞能稳定高表达LIGHT分子,能特异性地向荷瘤小鼠体内肿瘤组织趋化并抑制肿瘤的生长,这种体内抗肿瘤功能可能与促进T淋巴细胞IL-2、IFN-γ等细胞因子的分泌,改善局部免疫抑制环境有关。Objective To investigate a potential gene therapy approach by mesenchymal stem cells(MSCs)modified by LIGHT(homologous to lymphotoxins)gene and herpes simplex virus thymidine kinase(HSV-TK)gene in vivo.Methods pIRES2-LIGHT and HSV-TK-EGFP genes were co-transfected to mesenchymal stem cells(MSCs/LT),and mesenchymal stem cells transfected with empty vector(MSCs/mock)and HSV-TK-EGFP(MSCs/T)were used as controls.The expressions of LIGHT and HSV-TK-EGFP in MSCs were analyzed.In vivo migration assays were applied to observe the migration of MSCs/LT to tumor.The therapeutic effects of MSCs/LT were observed in breast tumor cell-based animal model.The cytokines secretion of T lymphocytes in tumor tissues were detected by ELISA.Results MSCs/LT highly expressed LIGHT.We found that MSCs/LT displayed a strong tropism for tumor tissues through in vivo migration assays.MSCs/LT treatment could effectively inhibit tumor growth in vivo,but after MSCs/LT apoptosis induced by ganciclovir for about 4 weeks,the tumor grew aggressively again.Compared with MSCs/mock and MSCs/T,MSCs/LT significantly increased the secretion of interferon-gamma(IFN-γ)and interleukin-2(IL-2),whereas IL-10 cytokines decreased in tumor tissues(all P<0.05).Conclusion Mesenchymal stem cells transfected by LIGHT and HSV-TK-EGFP genes exert anti-tumor effect through modulating T cell function.Together,our findings indicate that MSCs/LT can home into and deliver immune stimulating molecules to tumor tissues effectively,thus reversing the immune-suppressive environment and exerting anti-tumor immune effect.

关 键 词：抗肿瘤免疫 间充质干细胞 转染 LIGHT基因 HSV-TK-EGFP基因 

分 类 号：R730.3[医药卫生—肿瘤]