脂多糖激活Toll样受体4介导自噬促进食管鳞癌细胞增殖与细胞周期S期阻滞的研究

发布时间：2018-06-13 20:53

本文选题：脂多糖 + Toll样受体　；参考：《中国临床药理学杂志》2017年24期

【摘要】：目的检测Toll样受体4(TLR4)在食管鳞癌(ESCC)组织、癌旁组织、ESCC细胞系表达,脂多糖(LPS)在ESCC细胞系中对TLR4是否上调、以及对自噬、增殖、细胞周期等效应的影响。方法选取50例ESCC组织标本(ESCC组)和50例配对癌旁正常组织标本(癌旁正常组),用免疫组化法检测组织中TLR4表达特征和阳性表达情况。在ESCC细胞系Eca-109和TE-1中,1μg·mL~(-1)LPS处理作为实验组,不加药物刺激作为对照组,加入siRNA-TLR4作为si-TLR4组,加入siRNA-NC作为si-NC组,检测LPS对TLR4表达的影响,LPS、TLR4对自噬相关蛋白LC-3、ATG5、ATG7表达的影响;进一步用CCK8和流式细胞术分析siRNA干扰TLR4表达后对ESCC细胞增殖和细胞周期的影响。结果 TLR4在ESCC组和癌旁正常组的阳性表达率分别为68.00%和36.00%,差异有统计学意义(P0.01)。实验组和对照组的TLR4蛋白相对表达量在Eca-109细胞中分别为(0.11±0.05)和(0.35±0.12),在TE-1细胞中分别为(0.07±0.02)和(0.41±0.10),差异均有统计学意义(均P0.05)。LPS激活TLR4表达的同时能明显上调ESCC细胞系自噬相关蛋白LC-3、ATG5、ATG7的表达,用si-RNA敲降TLR4表达后,ESCC细胞系自噬相关蛋白LC-3、ATG5、ATG7的表达显著下调。敲降TLR4表达引起ESCC细胞周期S期停滞,si-NC组和si-TLR4组第5天细胞生存率在Eca-109细胞中分别为(1.35±0.02)和(0.92±0.04),在TE-1细胞中分别为(1.35±0.02)和(0.87±0.04),差异均有统计学意义(均P0.05)。结论 TLR4蛋白在ESCC组织过表达,LPS激活TLR4通路促进ESCC细胞自噬效应、进而促进ESCC发生发展。
[Abstract]:Objective to detect the expression of Toll-like receptor 4 (TLR4) in esophageal squamous cell carcinoma (ESCC) and its adjacent tissues. Whether lipopolysaccharide (LPS4) upregulated TLR4 and its effects on autophagy, proliferation and cell cycle in ESCC cells. Methods 50 ESCC specimens and 50 matched normal tissues (paracancerous normal tissues) were selected to detect the expression of TLR4 and the expression of TLR4 by immunohistochemical method. In ESCC cell lines Eca-109 and TE-1, 1 渭 g mLLP-1 lipopolysaccharide was treated as experimental group, without drug stimulation as control group, siRNA-TLR4 as si-TLR4 group and siRNA-NC as si-NC group. The effects of LPS on the expression of TLR4 and the expression of ATG5ATG7 in macrophage associated protein LC-3ATLR4 were detected. The effects of siRNA interference TLR4 expression on the proliferation and cell cycle of ESCC cells were further analyzed by CCK8 and flow cytometry. Results the positive expression rates of TLR4 in ESCC group and adjacent normal group were 68.00% and 36.00%, respectively. The difference was statistically significant (P 0.01). The relative expression of TLR4 protein in Eca-109 cells was 0.11 卤0.05) and 0.35 卤0.12 in Eca-109 cells, and 0.07 卤0.02 in TE-1 cells and 0.41 卤0.10 in TE-1 cells, respectively. The difference was statistically significant (all P0.05. LPS activated TLR4 expression in ESCC cell lines was significantly up-regulated at the same time. Expression of terminal protein LC-3, ATG5, ATG7, The expression of ATG7 in ESCC cell line was significantly down-regulated after TLR4 expression was reduced by si-RNA knockout. The 5th day survival rate of Eca-109 cells was 1.35 卤0.02 and 0.92 卤0.04, respectively, and that of TE-1 cells was 1.35 卤0.02 and 0.87 卤0.04, respectively. Conclusion LPS-activated TLR4 pathway in ESCC promotes autophagy of ESCC cells and promotes the development of ESCC.
【作者单位】：武汉大学人民医院老年病科;华中科技大学同济医学院附属同济医院胸外科;
【基金】：湖北省自然科学基金面上基金资助项目(2014CKB503)
【分类号】：R735.1

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