表没食子儿茶素没食子酸酯对人骨肉瘤细胞143B的抑制作用及其机制探讨

发布时间：2018-05-22 08:18

本文选题：骨肉瘤 + Wnt/β-catenin信号通路　；参考：《肿瘤》2017年06期

【摘要】：目的:研究表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对人骨肉瘤细胞143B的抑制作用,并初步探讨其可能的分子机制。方法:分别用0~50μmol/L的EGCG处理骨肉瘤143B细胞,然后采用结晶紫染色法观察细胞增殖能力的变化,Hoechst染色法分析细胞凋亡情况,细胞划痕愈合实验和Transwell小室法分析细胞迁移能力,蛋白质印迹法检测细胞凋亡及迁移相关蛋白的表达水平。同时,采用荧光素酶报告基因系统分析细胞中Wnt/β-catenin信号通路的活化情况,再用蛋白质印迹法检测Wnt/β-catenin信号通路中关键因子β-catenin及其下游靶分子c-Myc和cyclin D1的表达情况。结果:EGCG处理后,骨肉瘤143B细胞的增殖和迁移能力均受到明显抑制(P值均0.05),而晚期凋亡细胞数明显增加(P0.05);细胞中凋亡蛋白caspase-3及其剪切体的表达水平均明显上调(P值均0.05),而Bcl-2、金属基质蛋白酶2(matrix metalloproteinase-2,MMP-2)、β-catenin、c-Myc和cyclin D1的蛋白表达水平均明显下调(P值均0.05)。此外,EGCG处理组的荧光素酶活性较未处理组明显下降(P0.05)。结论:EGCG能抑制骨肉瘤143B细胞的增殖和迁移,促进细胞凋亡;这一作用可能与其阻断Wnt/β-catenin信号通路有关。
[Abstract]:Aim: to investigate the inhibitory effect of epigallocatechin-3-gallateEGCGG on human osteosarcoma cell line 143B and its possible molecular mechanism. Methods: osteosarcoma 143B cells were treated with 0 ~ 50 渭 mol/L EGCG, and cell proliferation was observed by crystal violet staining. Cell apoptosis was analyzed by Hoechst staining. Cell migration was analyzed by cell scratch healing assay and Transwell chamber assay. The expression of apoptosis and migration related proteins was detected by Western blot. Meanwhile, luciferase reporter gene system was used to analyze the activation of Wnt/ 尾 -catenin signaling pathway, and the expression of 尾 -catenin and its downstream target molecules, c-Myc and cyclin D1, in Wnt/ 尾 -catenin signaling pathway was detected by Western blot. Results after being treated with 10% EGCG, The proliferation and migration ability of osteosarcoma 143B cells were significantly inhibited (P = 0.05), while the number of late apoptotic cells increased significantly (P 0.05), and the expression of apoptotic protein caspase-3 and its shearing bodies increased significantly (P < 0.05), while Bcl-2 and metal group increased significantly the expression level of apoptotic protein caspase-3 and its shearing bodies in osteosarcoma 143B cells. The protein expression levels of 2(matrix metalloproteinase-2, 尾 -cateninine c-Myc and cyclin D1 were all significantly down-regulated (P < 0.05). In addition, the luciferase activity of EGCG group was significantly lower than that of untreated group. ConclusionEGCG can inhibit the proliferation and migration of osteosarcoma 143B cells and promote apoptosis, which may be related to the blocking of Wnt/ 尾 -catenin signaling pathway.
【作者单位】：重庆医科大学附属儿童医院骨科儿童发育疾病研究教育部重点研究室儿童发育重大疾病国家国际科技合作基地儿科学重庆重点实验室;
【基金】：国家临床重点专科建设项目(国卫办医涵[2013]544号)~~
【分类号】：R738.1

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