卡介苗诱导膀胱癌T24细胞表达钟声蛋白样受体和产生细胞因子的研究

Study on Toll-like receptors expression and cytokine production induced by bacillus Calmette-Guerin in human bladder cancer cell

目的 探讨卡介苗 (BCG)刺激对膀胱癌T2 4细胞钟声蛋白样受体 (TLR)表达和细胞因子产生水平的影响。方法 人膀胱癌细胞株T2 4用含 10 %FBS的RPMI16 4 0液培养 ,加入不同剂量(菌数 /细胞数比值 )梯度的BCG刺激。 1h后以RT PCR检测细胞TLR 2和TLR 4的表达水平 ,12h后ELISA法测定细胞上清液中IL 4和IL 12浓度。结果 BCG刺激后 ,T2 4细胞的TLR 2和TLR 4表达水平以及IL 12的分泌量随BCG剂量增加而升高 ,在菌数 /细胞数比值为 10时达最高 ;而IL 4分泌量与未刺激组相比无明显差别。结论 BCG刺激可使膀胱癌T2 4细胞TLR 2和TLR 4表达增强、IL 12水平升高 ,并呈剂量 效应关系 ;IL 12水平升高可能与TLR 2和TLR

Objective To investigate the effect of bacillus Calmette Guerin (BCG) on Toll like receptors (TLRs) expression and cytokine production in human bladder cancer cell line T24. Methods Human transiti onal carcinoma cell line T24 was cultured in RPMI1640 medium with 10% FBS, BCG was added into the cell culture with various doses in bacteria cell ratio. After T24 cells were stimulated by BCG for 1 hour, total RNA of cells was extracted. RT PCR procedure was conducted with the primer of TLR 2 and TLR 4, and the products were analyzed with agarose gels electrophoresis. Then the expression of TLR 2 and TLR 4 in T24 cells was assessed by the analyzing system of gel imaging. After the stimulation culture for 12 hours, the supernatant of cells was collected. The levels of IL 4 and IL 12 in each sample were assayed by ELISA method. Results The expression level of either TLR 2 or TLR 4 was increased by the stimulation of BCG in a dose dependent manner, the effects reached the maximal level at the dose of BCG as 10 bacilli per cell. The production of IL 12 in T24 cells was also increased gradually by the stimulation of BCG in a dose dependent manner, and the dose of BCG obtained maximal effect was 10 bacilli per cell, which is coincident with the results observed in the expression of TLRs. There was no difference showed on the production of IL 4 between T24 cells stimulated with BCG and control. Conclusions The stimulation of BCG not only up regulated the expression of TLR 2 and TLR 4, but also increased the production of IL 12 in bladder cancer T24 cell line. The expression of TLRs and the production of cytokine in bladder cancer cells may be related to the BCG induced immunol response to human bladder cancer.