脂多糖介导的TLR4信号通路在膀胱癌免疫逃逸中的作用及其机制

发布时间：2018-09-05 07:13

【摘要】：目的探讨脂多糖介导的Toll样受体(TLR)4信号通路活化在膀胱癌(BC)T24细胞系免疫逃逸中的作用及其机制。方法取对数生长期T24细胞,使用1μg/ml的脂多糖分别刺激该细胞0、6、12、24 h,采用流式细胞仪检测细胞表面TLR4的表达量,采用RT-PCR检测细胞中程序性死亡配体-1(PD-1,又称B7-H1)mRNA的表达,采用酶联免疫吸附法(ELISA)检测细胞中B7-H1蛋白的表达,分析TLR4表达与B7-H1 mRNA及其蛋白的相关性。结果 TLR4阳性率随刺激时间的增长而上调,且在12 h时达到最高值,24 h时表达略有降低,但仍较高。6、12、24 h时TLR4阳性率与0 h时比较差异有统计学意义(P0.05)。RT-PCR及ELISA结果显示B7-H1 mRNA及蛋白的表达随刺激时间的增长而上调,且在12 h时达到最高值,24 h时表达略有降低,但仍较高。与0 h时比较,6 h和24 h T24细胞B7-H1 mRNA及蛋白表达量增高(P0.05),12 h显著增高(P0.01)。TLR4与B7-H1 mRNA呈正相关(r=0.785,P=0.002),TLR4与B7-H1蛋白呈正相关(r=0.825,P=0.012)。结论脂多糖能活化TLR4信号通路,并上调B7-H1 mRNA及其蛋白的表达,可能参与BC细胞免疫逃逸,为BC的靶向治疗提供新思路。
[Abstract]:Objective to investigate the role and mechanism of lipopolysaccharide mediated activation of (TLR) 4 signaling pathway in bladder cancer (BC) T 24 cell line. Methods T24 cells in logarithmic growth stage were stimulated with 1 渭 g/ml lipopolysaccharide for 1224 h. The expression of TLR4 on the cell surface was detected by flow cytometry, and the expression of PD-1, ligand 1 (B7-H1) mRNA was detected by RT-PCR. The expression of B7-H1 protein was detected by enzyme linked immunosorbent assay (ELISA), and the correlation between TLR4 expression and B7-H1 mRNA and its protein was analyzed. Results the positive rate of TLR4 was up-regulated with the increase of stimulation time, and the expression of TLR4 decreased slightly at 12 h after reaching the highest value at 24 h. The results of RT-PCR and ELISA showed that the expression of B7-H1 mRNA and protein was up-regulated with the increase of stimulation time, and the expression of B7-H1 mRNA and protein decreased slightly at 12 h and reached the peak at 24 h, but it was still higher than that at 0 h. Compared with 0 h, the expression of B7-H1 mRNA and protein in T24 cells at 6 h and 24 h were significantly increased (P0.05). TLR4 was positively correlated with B7-H1 mRNA (r = 0.785P ~ (0.002) and B7-H1 protein (r ~ (0.825) P ~ (0.012). Conclusion lipopolysaccharide can activate TLR4 signaling pathway and up-regulate the expression of B7-H1 mRNA and its protein, which may participate in the escape of BC cells and provide a new idea for the targeted treatment of BC.
【作者单位】：江西省肿瘤医院泌尿外科;
【基金】：江西省自然科学基金资助项目(No.20161BAB205271)
【分类号】：R737.14

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