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学科主题基础医学
Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis
Pan, Yan1,2; Zhong, Li-jun3; Zhou, Hong1,2; Wang, Xin1,2; Chen, Kui1,2; Yang, Hao-peng1,2; Xiaokaiti, Yilixiati1,2; Maimaiti, Aikebaier1,2; Jiang, Ling1,2; Li, Xue-jun1,2
关键词heparin cell growth proteome vimentin 14-3-3 zeta/delta alpha(v)-integrin
刊名ACTA PHARMACOLOGICA SINICA
2012-06-01
DOI10.1038/aps.2012.42
33期:6页:798-808
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Multidisciplinary ; Pharmacology & Pharmacy
研究领域[WOS]Chemistry ; Pharmacology & Pharmacy
关键词[WOS]MOLECULAR-WEIGHT HEPARIN ; LUNG-CANCER ; ADHESION
英文摘要

Aim: To investigate the inhibitory effects of heparin on PC-3M cells proliferation in vitro and B16-F10-luc-G5 cells metastasis in Balb/c nude mice and identify the protein expression patterns to elucidate the action mechanism of heparin.

Methods: Human prostate cancer PC-3M cells were incubated with heparin 0.5 to 125 mu g/mL for 24 h. The proliferation of PC-3M cells was assessed by MTS assay. BrdU incoporation and Ki67 expression were detected using a high content screening (HCS) assay. The cell cycle and apoptosis of PC-3M cells were tested by flow cytometry. B16-F10-luc-G5 cardinoma cells were injected into the lateral tail vein of 6-week old male Balb/c nude mice and heparin 30 mg/kg was administered iv 30 min before and 24 h after injection. The metasis of B16-F10-luc-G5 cells was detected by bioluminescence assay. Activated partial thromboplastin time (APTT) and hemorheological parameters were measured on d 14 after injection of B16-F10-luc-G5 carcinoma cells in Balb/c mice. The global protein changes in PC-3M cells and frozen lung tissues from mice burdened with B16-F10-luc-G5 cells were determined by 2-dimensional gel electrophoresis and image analysis. The protein expression of vimentin and 14-3-3 zeta/delta was measured by Western blot. The mRNA transcription of vimentin, transforming growth factor (TGF)-beta, E-cadherin, and alpha(v)-integrin was measured by RT-PCR.

Results: Heparin 25 and 125 mu g/mL significantly inhibited the proliferation, arrested the cells in G(1) phase, and suppressed BrdU incorporation and Ki67 expression in PC-3M cells compared with the model group. But it had no significant effect on apoptosis of PC-3M cells. Heparin 30 mg/kg markedly inhibits the metastasis of B16-F10-luc-G5 cells on day 8. Additionally, heparin administration maintained relatively normal red blood hematocrit but had no influence on APTT in nude mice burdened with B16-F10-luc-G5 cells. Thirty of down-regulated protein spots were identified after heparin treatment, many of which are related to tumor development, extracellular signaling, energy metabolism, and cellular proliferation. Vimentin and 14-3-3 zeta/delta were identified in common in PC-3M cells and the lungs of mice bearing B16-F10-luc-G5 carcinoma cells. Heparin 25 and 125 mu g/mL decreased the protein expression of vimentin and 14-3-3 zeta/delta and the mRNA expression of alpha(v)-integrin. Heparin 125 mu g/mL decreased vimentin and E-cadherin mRNA transcription while increased TGF-beta mRNA transcription in the PC-3M cells, but the differences were not significant. Transfection of vimentin-targeted siRNA for 48 h significantly decreased the BrdU incoporation and Ki67 expression in PC-3M cells.

Conclusion: Heparin inhibited PC-3M cell proliferation in vitro and B16-F10-luc-G5 cells metastasis in nude mice by inhibition of vimentin, 14-3-3 zeta/delta, and alpha(v)-integrin expression.

语种英语
WOS记录号WOS:000305034500011
项目编号81020108031 ; 30572202 ; 30973558 ; 30772571 ; 30901815 ; 30901803 ; 2009ZX09103-144 ; B07001
资助机构National Natural Science Foundation of China ; Ministry of Science and Technology in China ; Ministry of Education of China ; Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry
引用统计
被引频次:8[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/63398
专题北京大学基础医学院_药理学系
北京大学基础医学院
北京大学医药卫生分析中心_研发办公室
北京大学第三临床医学院_超声影像科
作者单位1.Peking Univ, State Key Lab Nat & Biomimet Drugs, Dept Pharmacol, Sch Basic Med Sci, Beijing 100191, Peoples R China
2.Peking Univ, Inst Syst Biomed, Beijing 100191, Peoples R China
3.Peking Univ, Ctr Med Anal, Beijing 100191, Peoples R China
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Pan, Yan,Zhong, Li-jun,Zhou, Hong,et al. Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis[J]. ACTA PHARMACOLOGICA SINICA,2012,33(6):798-808.
APA Pan, Yan.,Zhong, Li-jun.,Zhou, Hong.,Wang, Xin.,Chen, Kui.,...&Li, Xue-jun.(2012).Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis.ACTA PHARMACOLOGICA SINICA,33(6),798-808.
MLA Pan, Yan,et al."Roles of vimentin and 14-3-3 zeta/delta in the inhibitory effects of heparin on PC-3M cell proliferation and B16-F10-luc-G5 cells metastasis".ACTA PHARMACOLOGICA SINICA 33.6(2012):798-808.
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