血小板衍生生长因子对脉络膜黑色素瘤细胞活性及侵袭能力的影响
Effect of platelet-derived growth factor on the activity and invasion ability of choroidal melanoma cells
姚宁宁 1袁莹莹 1马清悦 1易雯丹 1隋爱华 2罗文娟1
作者信息
- 1. 青岛大学附属医院眼科,山东青岛 266003
- 2. 青岛大学附属医院医学研究中心,山东青岛 266003
- 折叠
摘要
目的 探讨血小板衍生生长因子(PDGF)对脉络膜黑色素瘤(choroidal melanoma,CM)细胞活性及侵袭能力的影响.方法 将CM细胞分为A、B组,分别转染空载体和敲降PDGF的慢病毒,采用RT-qPCR技术检测两组细胞中PDGF mRNA相对表达量,采用XTT实验检测两组细胞活性,采用Transwell实验检测两组细胞侵袭能力,采用Western blot实验检测两组细胞上皮-间质转化(EMT)及基质金属蛋白酶(MMPs)相关标志物蛋白的相对表达量.GEPIA数据库分析PDGF水平与CM患者预后的关系.结果 与A组比较,B组细胞中PDGF RNA相对表达量下降(t=176.30,P＜0.05),细胞存活数目下降及侵袭能力减弱(F=57.21,t=14.10,P＜0.05).W estern blot实验结果显示,与A组相比,B组细胞中E-cadherin相对表达量升高,N-cadherin、Snail、Vimentin、MMP9 及 MT1 MMP 的相对表达量下降(t=4.13～14.14,P＜0.05).GEPIA 数据库分析显示,PDGF-A、PDGF-B高表达与CM患者的不良预后有关(P＜0.05).结论 PDGF具有增强CM细胞活性,促进CM细胞侵袭的作用,其机制可能与诱导细胞EMT的发生有关.
Abstract
Objective To investigate the effect of platelet-derived growth factor(PDGF)on the activity and invasion ability of choroidal melanoma(CM)cells.Methods CM cells were divided into groups A and B and were transfected with empty vector and PDGF knockdown lentivirus,respectively.RT-qPCR was used to measure the relative expression level of PDGF mR-NA;XTT assay was used to measure cell activity;Transwell assay was used to test the invasion ability of cells in the two groups;Western blot was used to measure the relative expression levels of epithelial-mesenchymal transition(EMT)marker proteins and matrix metalloproteinases(MMPs).GEPIA database was used to investigate the association between PDGF and the prognosis of CM patients.Results Compared with group A,group B had significant reductions in the relative expression level of PDGF RNA(t=176.30,P＜0.05),the number of cells which survived(F=57.21,P＜0.05),and the invasion ability of cells(F=57.21,t=14.10,P＜0.05).Western blot showed that compared with group A,group B had a significant increase in the relative ex-pression level of E-cadherin and significant reductions in the relative expression levels of N-cadherin,Snail,Vimentin,MMP9,and MT1 MMP(t=4.13-14.14,P＜0.05).The GEPIA database analysis showed that the high expression levels of PDGF-A and PDGF-B were associated with the poor prognosis of CM patients(P＜0.05).Conclusion PDGF can enhance the activity of CM cells and promote the invasion of CM cells,possibly by inducing EMT.
关键词
黑色素瘤/脉络膜肿瘤/血小板源性生长因子/细胞增殖/肿瘤浸润/上皮-间质转化/基质金属蛋白酶类Key words
Melanoma/Choroid neoplasms/Platelet-derived growth factor/Cell proliferation/Neoplasm invasiveness/Epithelial-mesenchymal transition/Matrix metalloproteinases引用本文复制引用
基金项目
山东省自然科学基金面上项目(82274585)
出版年
2024
国家科技期刊平台
NSTL
万方数据