Id1 enhances human ovarian cancer endothelial progenitor cell angiogenesis via PI3K/Akt and NF-κB/MMP-2 signaling pathways

Journal of Translational Medicine

Songbin Fu¹  Shiyun Peng³  Jialin Cui³  Ying Wang³  Lichen Teng³  Lingjuan Gao²  Yajuan Su³ 

[1] Laboratory of Medical Genetics, Harbin Medical University, Harbin, 150081, China;Department of clinical laboratory, Nanjing Maternity and Child Health Care hospital, Nanjing Medical University, Nanjing, China;Department of clinical laboratory, the affiliated tumor hospital, Harbin Medical University, Harbin, China

关键词: NF-κB/MMP-2;    PI3K/Akt;    Angiogenesis;    Endothelial progenitor cells;    Id1;   

Others  :  827510
DOI  :  10.1186/1479-5876-11-132

 received in 2013-03-11, accepted in 2013-05-22,  发布年份 2013

【 摘 要 】

Background

Endothelial progenitor cells (EPCs) contribute to tumor angiogenesis and growth. We previously reported that over-expression of an inhibitor of DNA binding/differentiation 1 (Id1) in EPCs can enhance EPC proliferation, migration, and adhesion. In this study, we investigated the role of Id1 in EPC angiogenesis in patients with ovarian cancer and the underlying signaling pathway.

Methods

Circulating EPCs from 22 patients with ovarian cancer and 15 healthy control subjects were cultured. Id1 and matrix metalloproteinase-2 (MMP-2) expression were analyzed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and western blot. EPC angiogenesis was detected by tube formation assays. Double-stranded DNA containing the interference sequences was synthesized according to the structure of a pGCSIL-GFP viral vector and then inserted into a linearized vector. Positive clones were identified as lentiviral vectors that expressed human Id1 short hairpin RNA (shRNA).

Results

Id1 and MMP-2 expression were increased in EPCs freshly isolated from ovarian cancer patients compared to those obtained from healthy subjects. shRNA-mediated Id1 down-regulation substantially reduced EPC angiogenesis and MMP-2 expression. Importantly, transfection of EPCs with Id1 in vitro induced phosphorylation of Akt (p-Akt) via phosphoinositide 3-kinase and increased the expression of MMP-2 via NF-κB. Blockage of both pathways by specific inhibitors (LY294002 and PDTC, respectively) abrogated Id1-enhanced EPC angiogenesis.

Conclusions

Id1 can enhance EPC angiogenesis in ovarian cancer, which is mainly mediated by the PI3K/Akt and NF-κB/MMP-2 signaling pathways. Id1 and its downstream effectors are potential targets for treatment of ovarian cancer because of their contribution to angiogenesis.

【 授权许可】

   
2013 Su et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]Kopp HG, Ramos CA, Rafii S: Contribution of endothelial progenitors and proangiogenic hematopoietic cells to vascularization of tumor and ischemic tissue. Curr Opin Hematol 2006, 13:175-181.
• [2]Asahara T, Takahashi T, Masuda H: VEGF contributes to postnatal neovascularization by mobilizing bone marrow derived endothelial progenitor cells. Embo J 1999, 18:3964-3972.
• [3]Urbich C, Dimmeler S: Endothelial progenitor cells: characterization and role in vascular biology. Circ Res 2004, 95:343-353.
• [4]Gao D, Nolan DJ, Mellick AS: EPCs control the angiogenic switch in mouse lung metastasis. Science 2008, 319:195-198.
• [5]Maw MK, Fujimoto J, Tamaya T: Overexpression of inhibitor of DNA-binding (ID)-1 protein related to angiogenesis in tumor advancement of ovarian cancers. BMC Cancer 2009, 9:430. BioMed Central Full Text
• [6]Lyden D, Young AZ, Zagzag D: Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts. Nature 2007, 104:1260-1265.
• [7]Jankovic V, Ciarrocchi A, Boccuni P: Id1 restrains myeloid commitment, maintaining the self-renewal capacity of hematopoietic stem cells. Proc Natl Acad Sci U S A 2007, 104:1260-1265.
• [8]Shaked Y, Ciarrocchi A, Franco M, Lee CR, Man S, Cheung AM, Hicklin DJ, Chaplin D, Foster FS, Benezra R, Kerbel RS: Therapy-induced acute recruitment of circulating endothelial progenitor cells to tumors. Science 2006, 313(5794):1785-1787.
• [9]Su YJ, Zheng L, Wang Q, Bao J, Zh C, Ailan L: The PI3K/ Akt pathway upregulates Id1 and integrin α4 to enhance recruitment of human ovarian cancer endothelial progenitor cells. BMC Cancer 2010, 10:459. BioMed Central Full Text
• [10]Su Y, Zheng L, Wang Q, Li W, Cai Z, Xiong S, Bao J: Quantity and clinical relevance of circulating endothelial progenitor cells in human ovarian cancer. J Exp Clin Cancer Res 2010, 29(1):27. BioMed Central Full Text
• [11]Tsuchiya K, Kim Y, Ondrey FG, Lin J: Characterization of a temper-ature-sensitive mouse middle ear epithelial cell line. Acta Otolaryngol 2005, 125:823-829.
• [12]Gao DC, Nolan D, McDonnell K, Vahdat L, Benezra R: Bone marrow-derived endothelial progenitor cells contribute to the angiogenic switch in tumor growth and metastatic progression. Biochim Biophys Acta 2009, 1796:33-40.
• [13]Nolan DJ, Ciarrocchi A, Mellick AS, Jaggi JS, Bambino K: Bone marrow-derived endothelial progenitor cells are a major determinant of nascent tumor neovascularization. Genes Dev 2007, 21:1546-1558.
• [14]Peters BA, Diaz LA, Polyak K: Contribution of bone marrow-derived endothelial cells to human tumor vasculature. Nat Med 2005, 11:261-262.
• [15]Duda DG, Cohen KS, Kozin SV: Evidence for incorporation of bone marrow-derived endothelial cells into perfused blood vessels in tumors. Blood 2006, 107:2774-2776.
• [16]Mieno S, Boodhwani M, Robich MP, Clements RT, Sodha NR, Sellke FW: Effects of diabetes mellitus on VEGF-induced proliferation response in bone marrow derived endothelial progenitor cells. J Card Surg 2010, 25(5):618-625.
• [17]Spring H, Schuler T, Arnold B, Hammerling GJ, Ganss R: Chemo-kines direct endothelia l progenitors int o tumor neovessels. Proc Natl Acad Sci USA 2005, 102:18111-18116.
• [18]Du R, Lu KV, Petritsch C, Liu P, Ganss R, Passegue E, Song H, Vandenberg S, Johnson RS, Werb Z: HIF1 α induces the recruitment of bone marrow–derived vascular modulatory cells to regulate tumor angiogenesis and invasion. Cancer Cell 2008, 13:206-220.
• [19]Li B, Sharpe EE, Maupin AB, Teleron AA, Pyle AL, Carmeliet P, Young PP: VEGF and PlGF promote adult vasculogenesis by enhancing EPC re-cruitment and vessel formation at the site of tumor neovascularization. FASEB J 2006, 20:1495-1497.
• [20]Yang L, Huang J, Ren X, Gorska AE, Chytil A, Aakre M, Carbone DP, Matrisian LM, Richmond A, Lin PC: Abrogation of TGFβ signal-ing in mammary carcinomas recruits Gr-1+CD11b+myeloid cells that promote metastasis. Cancer Cell 2008, 3:23-35.
• [21]Dutta S, Wang FQ, Wu HS, Mukherjee TJ, Fishman DA: The NF-κB pathway mediates lysophosphatidic acid (LPA)-induced VEGF signaling and cell invasion in epithelial ovarian cancer (EOC). Gynecol Oncol 2011, 23(1):129-137.
• [22]Lin J, Guan Z, Wang C, Feng L, Zheng Y, Caicedo E, Bearth E, Peng JR, Gaffney P, Ondrey FG: Inhibitor of differentiation 1 contributes to head and neck squamous cell carcinoma survival via the NF-kappaB/survivin and phosphoinositide 3-kinase/Akt signaling pathways. Clin Cancer Res 2010, 1;16(1):77-87.
• [23]Mern DS, Hoppe-Seyler K, Hoppe-Seyler F, Hasskarl J, Burwinkel B: Targeting Id1 and Id3 by a specific peptide aptamer induces E-box promoter activity, cell cycle arrest, and apoptosis in breast cancer cells. Breast Cancer Res Treat 2010, 124(3):623-633.