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| PKD3促进非小细胞肺癌A549细胞迁移的作用和机制 | |
| 引用本文： | 邹志鹏,曾方银,冯丽,柯志勇,Wang Q Jane,邓凡.PKD3促进非小细胞肺癌A549细胞迁移的作用和机制[J].第三军医大学学报,2010,32(15). |
| 作者姓名： | 邹志鹏 曾方银 冯丽 柯志勇 Wang Q Jane 邓凡 |
| 作者单位： | 1. 南方医科大学,细胞生物学教研室,广州,510515 2. 南方医科大学,南方医院检验科,广州,510515 3. 15260,USA,Pennsylvania,Pittsburgh, University of Pittsburgh School of Medicine, Department of Pharmacology and Chemical Biology |
| 基金项目： | 国家自然科学基金，教育部留学回国人员科研启动基金 |
| 摘 要： | 目的 探讨PKD3在非小细胞肺癌细胞迁移和侵袭中的作用及机制.方法 以人非小细胞肺癌A549细胞为研究对象,通过重组腺病毒感染和siRNA转染的策略,采用HEK293细胞测定病毒滴度;荧光定量RT-PCR和Western blot检测转染对照siRNA与PKD3 siRNA的A549细胞中PKD3、MT1-MMP、MMP-2的mRNA和蛋白表达水平差异;创伤愈合实验检测PKD3过表达或siRNA转染后不同时间点各组细胞的迁移能力.结果 经HEK293细胞病毒滴度测定,3种重组腺病毒Ade-LacZ、Ade-PKD3和Ade-PKD3-DN滴度分别达2.4×109、3.0×109、3.2×109 PFU/ml;免疫印迹证实Ade-PKD3和Ade-PKD3-DN在A549中高效过表达;在感染细胞24 h后和细胞划痕后4 、20 h,与Ade-LacZ感染的对照细胞相比,感染Ade-PKD3的A549迁移能力显著增强(P<0.01),而细胞划痕后8 h和20 h后,与Ade-LacZ感染细胞相比,Ade-PKD3-DN的A549迁移能力显著减弱(P<0.01);同时,siRNA敲低A549细胞中内源性PKD3的表达不仅在细胞划痕后的16、24、36 h明显抑制细胞的迁移,而且在mRNA和蛋白水平下调肿瘤细胞迁移和侵袭相关基因MT1-MMP、MMP-2的表达.结论 PKD3的表达可显著增强A549的迁移能力,这种作用可能通过上调MT1-MMP、MMP-2的表达实现. |
| 关 键 词： | 非小细胞肺癌 蛋白激酶D3 腺病毒 细胞迁移 |
Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells |
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| Zou Zhipeng,Zeng Fangyin,Feng Li,Ke Zhiyong,Wang Q Jane,Deng Fan.Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells[J].Acta Academiae Medicinae Militaris Tertiae,2010,32(15). | |
| Authors: | Zou Zhipeng Zeng Fangyin Feng Li Ke Zhiyong Wang Q Jane Deng Fan |
| Institution: | Zou Zhipeng1,Zeng Fangyin2,Feng Li1,Ke Zhiyong1,Wang Q Jane3,Deng Fan1(1Department of Cell Biology,2Department of Clinical Laboratory,Nanfang Hospital,Southern Medical University,Guangzhou,510515,Guangdong Province,China,3Department of Pharmacology and Chemical Biology,University of Pittsburgh School of Medicine,Pittsburgh,Pennsylvania,15260,USA) |
| Abstract: | Objective To study the role and mechanism of protein kinase D3(PKD3) in promoting migration and invasion of non-small cell lung cancer(NSCLC) cells.Methods Human NSCLC A549 cells were infected with a high titer of Ade-LacZ,Ade-PKD3 or Ade-PKD3-DN adenovirus or transfected with siRNA of non-specific control(si-CTL) and siRNA of PKD3(si-PKD3),respectively.Expression of PKD3,MT1-MMP,MMP-2 mRNAs and proteins in NSCLC A549 cells was detected by real-time RT-PCR and Western blot analysis,respectively.Migration of... |
| Keywords: | siRNA |
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