靶向沉默膀胱癌T24细胞系VEGF对树突状细胞分化成熟及免疫功能的影响

Influence of targeted gene VEGF silencing of bladder cancer cell line T24 on differentiation,maturation and function of dendritic cells

目的观察靶向沉默膀胱癌T24细胞系VEGF对树突状细胞(DC)分化成熟及免疫功能的影响。方法构建VEGF慢病毒干扰载体LV-VEGFA-RNAi(实验组)及慢病毒阴性对照载体LV-CON(阴性对照组),分别感染各组T24细胞,空白对照组不采取任何干预措施。分别用RT-PCR和ELISA检测各组T24细胞VEGF mRNA和蛋白的表达。各组T24上清分别与PBMC来源的未成熟DC共培养,用流式细胞仪检测DC分化成熟指标CD1a、CD83及免疫功能指标CD86。结果与空白对照组和阴性对照组相比,实验组T24细胞VEGF mRNA及蛋白的表达均受到显著抑制(P<0.05)。实验组DC表型CD1a、CD83和CD86较空白对照组明显增高(P<0.05),较阴性对照组亦明显增高(P<0.05)。结论靶向沉默膀胱癌T24细胞VEGF的表达,有利于微环境中DC的成熟及其免疫功能的发挥,可能通过修复DC被损害的免疫监视功能,增加DC的抗肿瘤能力。

Objective To observe the influence of targeted gene VEGF silencing of bladder cancer cell line T24 on differentiation,maturation and function of dendritic cells. Methods A lentiviral vector named LV-VEGFA-RNAi（ experimental group） for gene silencing targeting VEGF and a lentiviral vector named LV-CON（ negative control group） without any valid sequences were constructed. The blank control group accepted no intervention measures.The expression of VEGF＇s mRNA and protein of T24 cells from each group were detected by RT-PCR and ELISA respectively. Then the immature DCs were co-cultured respectively with the supernatant of all the groups as mentioned above. CD1 a,CD83 as the maturation marker and CD86 as the immunity marker of the DCs were detected by flow cytometry. Results The expression of VEGF＇s mRNA and protein of T24 cells in the experimental group were obviously inhibited（ P 0. 05） as compared with that in the negative control group and the blank control group. DCs of the experimental group had an obviously increased（ P 0. 05） expression of CD1 a,CD83 and CD86 compared with the negative control group and the blank control group. Conclusions Targeted gene VEGFsilencing by RNAi has advantages to the growth and immunity of DCs,which may strengthen the anti-tumor capacity of the DCs by repairing their damaged immune monitoring function.