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PKD3促进非小细胞肺癌A549细胞迁移的作用和机制 被引量：2

Role and mechanism of protein kinase D3 in promoting migration of non-small cell lung cancer A549 cells
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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Westernblot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、3.2×109PFU/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。 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-LaeZ, Ade-PKD3 or Ade-PKD3-DN adenovirns or transfeeted with siRNA of non-specific control （si-CTL） and siRNA of PKD3 （si-PKD3） , respectively. Expression of PKD3, MTI- 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 transfeeted 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 ×10^9, 3.0 ×10^9, and 3.2×10^9PFU/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-LaeZ in 4 and 20 h after cell scratching （P 〈 0. 01 ）, whereas dramatically reduced after infection with Ade-PKD3-DN in 8 and 20 h after cell scratching （ P 〈 0. 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 .
作者 邹志鹏 曾方银 冯丽 柯志勇 Wang Q Jane 邓凡
机构地区 南方医科大学:细胞生物学教研室 南方医科大学:南方医院检验科 University of Pittsburgh School of Medicine
出处 《第三军医大学学报》 CAS CSCD 北大核心 2010年第15期1609-1612,共4页 Journal of Third Military Medical University
基金 国家自然科学基金(30973014) 教育部留学回国人员科研启动基金(K1010353)~~
关键词 非小细胞肺癌 蛋白激酶D3 腺病毒 SIRNA 细胞迁移 non-small cell lung cancer protein kinase D3 adenovirus siRNA cell migration
分类号 R345.557 [医药卫生—基础医学] R730.23 [医药卫生—肿瘤]
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参考文献12

  • 1张明慧,胡义德,仲晨.含顺铂的4组联合化疗方案治疗晚期非小细胞肺癌的临床疗效比较[J].第三军医大学学报,2009,31(8):744-747. 被引量：11
  • 2Pallis A G, Serfass L, Dziadziusko R, et al. Targeted therapies in the treatment of advanced/metastatic NSCLC[J]. EurJ Cancer, 2009, 45 ( 14), 2473 -2487.
  • 3Gettinger S. Targeted therapy in advanced non-small-cell lung cancer [J]. Semin Respir Crit Care Med, 2008, 29(3) : 291 -301.
  • 4Ardizzoni A, Tiseo M. Novel targeted approaches in non-snmll cell lung cancer (NSCLC) [J]. Ann Oncol, 2006, 17( Suppl 5 ) : v91 - v93.
  • 5Chen J, Deng F, Singh S V, et al. Protein kinase D3 (PKD3) contributes to prostate cancer cell growth and survival thruugh a PKCepsi- Ion/PKD3 pathway downstream of Akt and ERK 1/2 [ J ]. Cancer Res, 2008, 68(10) : 3844-3853.
  • 6Lu G, Chen J, Espinoza L A, et al. Protein kinase D 3 is localized in vesicular structures and interacts with vesiele-assoeiated membrane protein 2[J]. Cell Signal, 2007, 19(4) : 867 -879.
  • 7Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C (t)) method [J]. Methods, 2001, 25(4): 402-408.
  • 8Anderson G, Chen J, Wang Q J. Individual C1 domains of PKD3 in phorbol ester-induced plasma membrane translocation of PKD3 in intact cells[ J]. Cell Signal, 2005, 17( 11 ) : 1397 - 1411.
  • 9Sahai E. Mechanisms of cancer cell invasion [ J ]. Curr Opin Genet Dev, 2005, 15(1): 87-96.
  • 10Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment [ J]. Cell, 2010, 141 ( 1 ) : 52 -67.

二级参考文献13

  • 1Carney D N. Lung cancer-time to move on from chemotherapy[J]. N Engl J Med, 2002, 346(2) : 126 -128.
  • 2Raez L E, Lilenbaum R. New developments in chemotherapy for advanced non-small cell lung cancer[J]. Curr Opin Oncol, 2006, 18 (2) : 156 - 161.
  • 3Laskin J J, Sandler A B. First-line treatment for advanced non-small- cell lung cancer [ J ]. Oncology ( Williston-Park ), 2005, 19 ( 13 ) : 1671 - 1676, 1678 - 1680.
  • 4Einhorn L H. First-line chemotherapy for non-small-cell lung cancer: is there a superior regimen based on histology? [ J]. J Clin Oncol, 2008, 26(21) : 3485 -3486.
  • 5Einhorn L H. Will we ever have personalized medicine for non-small cell lung cancer? [J]. J Thorac Oncol, 2006, 1 (7) : 737 -739.
  • 6Hsu D S, Balakumaran B S, Acharya C R, et al. Pharmacogenomic strategies provide a rational approach to the treatment of cisplatin-resistant patients with advanced cancer[ J]. J Clin Oncol, 2007, 25 (28) : 4350 - 4357.
  • 7Ruckdeschel J C, Finkelstein D M, Ettinger D S, et al. A randomized trial of the four most active regimens for metastatic non-small-cell lung cancer [J]. J Clin Oncol, 1986, 4(1): 14-22.
  • 8Sandier A, Gray R, Perry M C, et al. Paclitaxel-carboplatin alone or with bevacizumab for non-small-cell lung cancer [ J]. N Engl J Med, 2006, 355 (24) : 2542 - 2550.
  • 9Scagliotti G V, Parikh P, von-Pawel J, et al. Phase Ⅲ study comparing cisplatin plus gemeitabine with cisplatin plus pernetrexed in chemotherapy-naive patients with advanced-stage non-small-cell lung cancer[J]. J Clin Oncol, 2008, 26(21): 3543-3551.
  • 10Schiller J H, Harrington D, Belani C P, et al. Comparison of four chemotherapy regimens for advanced non-small-cell lung cancer [J]. N Engl J Med, 2002, 346(2) : 92 -98.

共引文献10

同被引文献16

  • 1Pienta K J, Bradley D. Mechanisms underlying the development of androgen-independent prostate cancer[J]. Clin Cancer Res, 2006, 12 (6) : 1665 -1671.
  • 2Sumitomo M, Ohba M, Asakuma J, et al. Protein kinase Cdelta amplifies ceramide formation via mitochondrial signaling in prostate cancer cells[ J]. J Clin Invest, 2002, 109 (6) : 827 - 836.
  • 3Salido M, Gonzalez J L, Vilches J. Loss of mitochondrial membrane potential is inhibited by bombesin in etoposide-induced apoptosis in PC-3 prostate carcinoma cells [ J ]. Mol Cancer Ther, 2007, 6 (4) : 1292 - 1299.
  • 4Uzzo R G, Haas N B, Crispen P L, et al. Mechanisms of apoptosis resistance and treatment strategies to overcome them in hormnone-refractmy prostate cancer[J]. Cancer, 2008, 112(8): 1660-1671.
  • 5Albihn A, Loven J, Ohlsson J,et al. c-Myc-dependent etoposide-induced apoptosis involves activation of Bax and caspases, and PKCdeha signaling[J]. J Cell Bioehem, 2006,98(6) : 1597 -1614.
  • 6Salido M, Larran J, Lopez A, et al. Etoposide sensitivity of human prostatic cancer cell lines PC-3, DU-145 and LNCaP[J]. Histol Histopathol, 1999, 14(1): 125-134.
  • 7Liu S, Yamauchi H. Etoposide induces growth arrest and disrupts androgen receptor signaling in prostate cancer cells [ J ]. Oncol Rep, 2010, 23(1) : 165 -170.
  • 8Wang Q J. PKD at the crossroads of DAG and PKC signaling [J].Trends Pharmacol Sei, 2006, 27 (6) : 317 - 323.
  • 9Rozengurt E, Rey O, Waldron R T. Protein kinase D signaling[J]. J Biol Chem, 2005, 280(14) : 13205 - 13208.
  • 10Chen J, Deng F, Singh S V, et al. Protein kinase D3 (PKD3) contributes to prostate cancer cell growth and survival through a PKCepsilon/PKD3 pathway downstream of Akt and ERK 1/2 [J]. Cancer Res, 2008, 68(10) : 3844 -3853.

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第三军医大学学报
2010年 第15期

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