Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells

邹志鹏; 曾方银; 冯丽; 柯志勇; Wang Q Jane; 邓凡

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Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells

更新时间：2026-01-09

- Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells
- Journal of Army Medical University Vol. 32, Issue 15, Pages: 1609-1612(2010)
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  CLC： R345.557;R730.23;R734.2
- Published：2010
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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].陆军军医大学学报(原第三军医大学学报),2010,32(15):1609-1612.

　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].J Amry Med Univ (J Third Mil Med Univ),2010,32(15):1609-1612.
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].陆军军医大学学报(原第三军医大学学报),2010,32(15):1609-1612. DOI：

　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].J Amry Med Univ (J Third Mil Med Univ),2010,32(15):1609-1612. DOI：

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2.4×10

、3.0×10

、3.2×10

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（P0.01），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（P0.01）；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。

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. Exp

ression 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 A549 cells infected with adenovirus or transfected with siRNA was detected at different time points by wound healing assay. Results The titer of Ade-LacZ

Ade-PKD3 or Ade-PKD3-DN adenovirus reached 2.4×10

3.0×10

and 3.2×10

PFU/ml

respectively. Western blotting showed that the PKD3 or dominant negative mutant PKD3 (PKD3-DN) was over-expressed. The migration level of A549 cells infected with Ade-PKD3 was significantly higher than that of those infected with Ade-LacZ in 4 and 20 h after cell scratching (P0.01)

whereas dramatically reduced after infection with Ade-PKD3-DN in 8 and 20 h after cell scratching (P0.01). Furthermore

PKD3 knocked down by siRNA transfection not only inhibited the migration of A549 cells in 16

24 and 36 h after cell scratch

but also down-regulated the expression of MT1-MMP

MMP-2 at mRNA and protein levels. Conclusion Expression of PKD3 can significantly increase the migration of NSCLC A549 cells by up-regulating the expressions of MT1-MMP and MMP-2 .

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