發(fā)布時間：2018-09-05 07:13

【摘要】：目的探討脂多糖介導的Toll樣受體(TLR)4信號通路活化在膀胱癌(BC)T24細胞系免疫逃逸中的作用及其機制。方法取對數(shù)生長期T24細胞,使用1μg/ml的脂多糖分別刺激該細胞0、6、12、24 h,采用流式細胞儀檢測細胞表面TLR4的表達量,采用RT-PCR檢測細胞中程序性死亡配體-1(PD-1,又稱B7-H1)mRNA的表達,采用酶聯(lián)免疫吸附法(ELISA)檢測細胞中B7-H1蛋白的表達,分析TLR4表達與B7-H1 mRNA及其蛋白的相關性。結果 TLR4陽性率隨刺激時間的增長而上調(diào),且在12 h時達到最高值,24 h時表達略有降低,但仍較高。6、12、24 h時TLR4陽性率與0 h時比較差異有統(tǒng)計學意義(P0.05)。RT-PCR及ELISA結果顯示B7-H1 mRNA及蛋白的表達隨刺激時間的增長而上調(diào),且在12 h時達到最高值,24 h時表達略有降低,但仍較高。與0 h時比較,6 h和24 h T24細胞B7-H1 mRNA及蛋白表達量增高(P0.05),12 h顯著增高(P0.01)。TLR4與B7-H1 mRNA呈正相關(r=0.785,P=0.002),TLR4與B7-H1蛋白呈正相關(r=0.825,P=0.012)。結論脂多糖能活化TLR4信號通路,并上調(diào)B7-H1 mRNA及其蛋白的表達,可能參與BC細胞免疫逃逸,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】：江西省自然科學基金資助項目(No.20161BAB205271)
【分類號】：R737.14

【相似文獻】

相關期刊論文 前5條

1 聶祥智;朱忠華;劉建設;楊曉;;脂多糖促進人近端腎小管上皮細胞TLR4的表達[J];武漢大學學報(醫(yī)學版);2010年02期

2 余曉東;朱平宇;魯棟梁;唐鐵龍;楊雪松;;TLR4蛋白在腎缺血再灌注損傷中的表達及意義[J];川北醫(yī)學院學報;2009年06期

3 李映川;細胞毒T細胞功能缺陷與腎癌的免疫逃逸[J];國外醫(yī)學.泌尿系統(tǒng)分冊;2003年04期

4 伍宏亮;張青川;;對TLR4的認識及其與前列腺癌研究進展[J];中華全科醫(yī)學;2013年03期

5 廖新惠;謝輝;陳麗君;;TLR4在前列腺癌中的表達及臨床意義[J];臨床泌尿外科雜志;2011年10期

相關會議論文 前1條

1 徐凱;郭劍明;張沂南;劉宇軍;徐志兵;朱延軍;王國民;;冬凌草甲素對雄激素非依賴性前列腺癌裸鼠移植瘤免疫逃逸相關蛋白的影響[A];第十五屆全國泌尿外科學術會議論文集[C];2008年

相關博士學位論文 前2條

1 張新濤;前列腺干細胞抗原（PSCA）基因rs2294008多態(tài)性與膀胱癌易感性的關聯(lián)研究[D];安徽醫(yī)科大學;2017年

2 華東;TLR4信號通路介導的前列腺癌侵襲轉移的研究[D];蘇州大學;2011年

相關碩士學位論文 前3條

1 白靜;氯沙坦對大鼠腎缺血再灌注損傷中TLR4表達的影響[D];山西醫(yī)科大學;2014年

2 紀傳彪;從T細胞免疫應答的初始階段研究TLR-4和B7-H1在膀胱癌免疫逃逸中的作用及臨床意義[D];青島大學;2013年

3 張鑫;軟脂酸對腎小管上皮細胞TLR4表達的影響[D];山西醫(yī)科大學;2014年

，

本文編號：2223563