摘要
目的:观察转染4-1BBL重组腺病毒载体的小鼠前列腺癌细胞瘤苗体外诱导抗肿瘤免疫作用及机制。方法:利用复制缺陷型腺病毒AdEasy-1系统,通过基因重组技术构建Ad-m4-1BBL及Ad-eGFP。将Ad-m4-1BBL和Ad-eGFP感染小鼠前列腺癌细胞RM-1,以丝裂霉素C(MMC)处理,制成肿瘤细胞疫苗(TCV,TCV-Ad-eG-FP,TCV-Ad-m4-1BBL),经体外与同系小鼠脾淋巴细胞共同培养,ELISA法检测脾细胞分泌细胞因子(IL-2,INF-γ)的影响,CCK-8试剂检测脾淋巴细胞特异性杀伤活性。结果:转染4-1BBL基因的小鼠前列腺癌细胞瘤苗TCV-Ad-m4-1BBL高表达4-1BBL蛋白,培养上清中细胞因子IL-2[(180.24±2.22)pg/ml]和INF-γ[(1 512.46±23.64)pg/ml]表达水平均明显高于TCV和TCV-Ad-eGFP组(P<0.05),TCV-Ad-m4-1BBL瘤苗诱导RM-1细胞特异性杀伤率[(34.24±2.64)%],相比TCV-Ad-eGFP[(14.65±3.21)%]和TCV[(9.82±1.48)%],差异有显著性(P<0.05)。但针对Hepal-6细胞,3种瘤苗均无效。结论:表达4-1BBL的小鼠前列腺癌细胞瘤苗能诱导有效的抗肿瘤免疫反应。
Objective：To explore the anti-tumor immunity in vitro induced by prostate cancer cell vaccine transfected with recombinant adenovirus encoding 4-1BBL in mice.Methods：The replication-deficient adenovirus AdEasy-1 system was used to construct recombinant adenovirus Ad-m4-1BBL and Ad-eGFP.The prostate cancer cell RM-1 of mice was transfected with Ad-m4-1BBL and Ad-eGFP,and treated with mitomycin（MMC） to produce TCV,TCV-Ad-eGFP and TCV-Ad-m4-1BBL,followed by co-culture with syngeneic murine spleen cells.Then the cytotoxic activity of the lymphocytes against RM-1 cells was analyzed with CCK-8 solution,and IL-2 and INF-γ were detected by ELISA.Results：The 4-1BBL protein was highly expressed in the TCV-Ad-m4-1BBL of the 4-1BBL-transfected mice.TCV-Ad-m4-1BBL significantly increased the expressions of IL-2（[180.24 ± 2.22] pg/ml） and INF-γ（[1 512.46 ± 23.64] pg/ml） as compared with TCV and TCV-Ad-eGFP（P〈0.05）,and induced higher RM-1 cell specific cytotoxicity（[34.24 ± 2.64]%） than the latter two（[9.82 ± 1.48]%） and（[14.65 ± 3.21]%）,（P〈0.05）.But none of them exhibited significant cytotoxicity against hepatocellular carcinoma Hepal-6.Conclusion：The m4-1BBL-expressing prostate cancer cell vaccine can effectively induce anti-tumor immune responses.Natl J Androl,2010,16（9）：773-777
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2010年第9期773-777,共5页
National Journal of Andrology
基金
国家自然科学基金(30672107)~~
