發(fā)布時(shí)間：2018-09-05 07:13

【摘要】：目的探討脂多糖介導(dǎo)的Toll樣受體(TLR)4信號(hào)通路活化在膀胱癌(BC)T24細(xì)胞系免疫逃逸中的作用及其機(jī)制。方法取對(duì)數(shù)生長(zhǎng)期T24細(xì)胞,使用1μg/ml的脂多糖分別刺激該細(xì)胞0、6、12、24 h,采用流式細(xì)胞儀檢測(cè)細(xì)胞表面TLR4的表達(dá)量,采用RT-PCR檢測(cè)細(xì)胞中程序性死亡配體-1(PD-1,又稱B7-H1)mRNA的表達(dá),采用酶聯(lián)免疫吸附法(ELISA)檢測(cè)細(xì)胞中B7-H1蛋白的表達(dá),分析TLR4表達(dá)與B7-H1 mRNA及其蛋白的相關(guān)性。結(jié)果 TLR4陽(yáng)性率隨刺激時(shí)間的增長(zhǎng)而上調(diào),且在12 h時(shí)達(dá)到最高值,24 h時(shí)表達(dá)略有降低,但仍較高。6、12、24 h時(shí)TLR4陽(yáng)性率與0 h時(shí)比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)。RT-PCR及ELISA結(jié)果顯示B7-H1 mRNA及蛋白的表達(dá)隨刺激時(shí)間的增長(zhǎng)而上調(diào),且在12 h時(shí)達(dá)到最高值,24 h時(shí)表達(dá)略有降低,但仍較高。與0 h時(shí)比較,6 h和24 h T24細(xì)胞B7-H1 mRNA及蛋白表達(dá)量增高(P0.05),12 h顯著增高(P0.01)。TLR4與B7-H1 mRNA呈正相關(guān)(r=0.785,P=0.002),TLR4與B7-H1蛋白呈正相關(guān)(r=0.825,P=0.012)。結(jié)論脂多糖能活化TLR4信號(hào)通路,并上調(diào)B7-H1 mRNA及其蛋白的表達(dá),可能參與BC細(xì)胞免疫逃逸,為BC的靶向治療提供新思路。
[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.
【作者單位】： 江西省腫瘤醫(yī)院泌尿外科;
【基金】：江西省自然科學(xué)基金資助項(xiàng)目(No.20161BAB205271)
【分類號(hào)】：R737.14

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