TSP-1基因对骨肉瘤血管生成的影响及其作用机制

doi:10.11904/j.issn.1002-3070.2019.02.002

实用肿瘤学杂志 ›› 2019, Vol. 33 ›› Issue (2): 103-109.doi: 10.11904/j.issn.1002-3070.2019.02.002

TSP-1基因对骨肉瘤血管生成的影响及其作用机制

朱欢叶¹, 简月奎², 李波², 高淦², 田震², 舒嫚²

1.贵州医科大学(贵阳 550004);
2.贵州省人民医院骨科

收稿日期:2018-08-14 修回日期:2018-12-11 出版日期:2019-04-20 发布日期:2019-04-25
通讯作者: 李波,E-mail:libo3@medmail.com.cn
作者简介:朱欢叶,男,(1993-),硕士,住院医师,从事脊柱外科的研究
基金资助:
贵州省留学人员科技创新项目(编号:201502); 贵州省人民医院国家自然科学基金补助基金项目(编号:黔科合平台人才[2017]5724-5)

Effect of TSP-1 gene on angiogenesis of osteosarcoma and its mechanism

ZHU Huanye¹, JIAN Yuekui², LI Bo², GAO Gan², TIAN Zhen², SHU Man²

1.Guizhou Medical University, Guiyang 550004, China;
2 Department of Orthopedics, Guizhou Provincial People's Hospital

Received:2018-08-14 Revised:2018-12-11 Online:2019-04-20 Published:2019-04-25

摘要/Abstract

摘要： 目的本研究旨在探讨TSP-1基因对人骨肉瘤血管生成的影响及其作用机制。方法通过构建pBPLV-shRNA-TSP-1干扰载体和pBPLV-TSP-1表达载体,对MG-63细胞进行转染,利用CCK8检测细胞活力,并测定其侵袭能力,然后通过免疫荧光检测细胞内CD36等表达量,qRT-PCR和Western blot检测细胞内TSP-1、CD36、p38MAPK、VEGF、VEGFR-1、EGF和PDGF等基因和蛋白的表达情况。结果 CCK-8及细胞侵袭实验结果显示转染TSP-1过表达载体后细胞活力和侵袭能力相较于空载体组均有显著增强(P＜0.05),而转染TSP-1干扰载体后细胞活力和侵袭能力均有明显降低(P＜0.05);免疫荧光、qPCR、Western blot结果显示过表达TSP-1后TSP-1、EGF、P38、PDGF、VEGF、VEGFR-1的mRNA水平及蛋白表达均有明显升高(P＜0.05),而转染TSP-1干扰载体后相关基因和蛋白的表达均有显著降低(P＜0.05)。结论 TSP-1基因能够促进MG-63细胞的增殖和侵袭能力,促进人骨肉瘤血管形成,其机制与生长因子EGF、VEGF、PDGF表达升高及P38-MAPK通路激活有关。

关键词: 骨肉瘤, TSP-1基因, MG-63, P38-MAPK通路

Abstract: Objective The aim of this study was to investigate the effect of TSP-1 gene on angiogenesis in human osteosarcoma and its mechanism of action. Methods MG-63 cells were transfected with constructing pBPLV-shRNA-TSP-1 vector and pBPLV-TSP-1 expression vector.Cell viability was measured by CCK8,and its invasive ability was measured by Transwell assay.The expression of CD36 in intracells was detected by immunofluorescence.The expression levels of TSP-1,CD36,p38MAPK,VEGF,VEGFR-1,EGF and PDGF were detected in MG-63 cells by qRT-PCR and Western blot. Results The cell viability and invasion ability were significantly increased after transfected pBPLV-TSP-1 vector compared with the empty vector group(P＜0.05),and significantly decreased after transfected pBPLV-shRNA-TSP-1 vector(P＜0.05).The expression of TSP-1,EGF,P38,PDGF,VEGF and VEGFR-1 at mRNA and protein levels was significantly increased after transfection pBPLV-TSP-1(P＜0.05),and significantly decreased after transfection pBPLV-shRNA-TSP-1 vector(P＜0.05). Conclusion TSP-1 gene can promote the proliferation and invasion of MG-63 cells,and promote the formation of human osteosarcoma,indicating its mechanism related to the increase of growth factors EGF,VEGF,PDGF and activation of P38-MAPK pathway.

Key words: Osteosarcoma, TSP-1, MG-63, P38-MAPK pathway

中图分类号:

R73-37

朱欢叶, 简月奎, 李波, 高淦, 田震, 舒嫚. TSP-1基因对骨肉瘤血管生成的影响及其作用机制[J]. 实用肿瘤学杂志, 2019, 33(2): 103-109.

ZHU Huanye, JIAN Yuekui, LI Bo, GAO Gan, TIAN Zhen, SHU Man. Effect of TSP-1 gene on angiogenesis of osteosarcoma and its mechanism[J]. PRACTICAL ONCOLOGY JOURNAL, 2019, 33(2): 103-109.

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TSP-1基因对骨肉瘤血管生成的影响及其作用机制

Effect of TSP-1 gene on angiogenesis of osteosarcoma and its mechanism

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