Research Article | Volume: 4, Issue: 2, March-April, 2016

Down-regulation of Id1 and Id3 genes affects growth and survival of Human Umbilical Vein Endothelial Cells (HUVECs)

Foster Kyei Du-Bois Asante Juliet Ama Mawusi Edekor Esther Sarpong Edem Gavor Daniels Konja   

Open Access   

Published:  Apr 21, 2016

DOI: 10.7324/JABB.2016.40204
Abstract

Inhibitors of DNA (Id) proteins have been implicated in cell growth, cell cycle control, apoptosis and angiogenesis. Studies reveal that Id1 and Id3 may be required for angiogenesis in adult tissues. This work aimed at investigating the consequences of loss-of-function of Id protein on growth and apoptosis in an in vitro model using Human Umbilical Vein Endothelial Cells (HUVECs). Using automated image analysis software (CellProfiler Analyst), cell cycle profiles of cell populations following siRNA down-regulation of Id proteins were investigated. Also, cell viability analysis with 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was evaluated. The study also employed flow cytometry using mitochondrial membrane-potential marker (3, 3’-Dihexyloxacarbocyanine Iodide) (DiOC6) and Propidium Iodide (PI) as a marker of non-viable necrotic cells. Down-regulation of Id1 and Id3 reduced viable HUVECs populations. In addition, knock down of Id1 and Id3 showed cell cycle arrest. Furthermore, cell death analysis following the silencing of Id1 and Id3 induced apoptosis. These findings suggest that the inhibition of Id1 and Id3 may impair endothelial cell activation and angiogenesis and may also provide an attractive target for the development of new therapies for angiogenic disorders.


Keyword:     Apoptosis Angiogenesis S-phase Down-regulation Human umbilical vein endothelial cells Knock-down.


Citation:

Kyei F, Asante DB, Edekor JAM, Sarpong E, Gavor E, Konja D. Down-regulation of Id1 and Id3 genes affects growth and survival of Human Umbilical Vein Endothelial Cells (HUVECs). J App Biol Biotech. 2016; 4 (02): 023-029. DOI: 10.7324/JABB.2016.40204

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

HTML Full Text
Reference

1. Norton JD. Id helix-loop-helix proteins in cell growth, differentiation and tumourigenesis. Journal of Cell Science. 2000; 113:3897-3905.

2. Hasmat A, Skider HS, Meghann K. Id proteins in cell growth and tumourigenesis. Cancer Cell. 2003; 6:525-530.

3. Massari ME, Murre C. Helix-loop-helix proteins: regulators of transcription in eukaryotic organisms. Molecular Cellular Biology. 2000; 20:429-440.

4. Lin CQ, Singh J, Murata K, Itahana Y, Parrinello S, Liang SH, et al. A role for Id1 in the aggressive phenotype and steroid hormone response of human breast cancer cells. Cancer Research. 2000; 60:1332–1340.

5. Iavarone A, Lasorella A. Id proteins as targets in cancer and tools in neurobiology. Trends in Molecular Medicine. 2006; 12:588-594.

6. Qiu J, Wang G, Peng Q, Hu J, Luo X, Zheng Y, et al. Id1 induces tubulogenesis by regulating endothelial cell adhesion and cytoskeletal organization through β1-integrin and Rho-kinase signalling. Cancer Letters. 2011; 307:191-199.

7. Sakurai D, Tsuchiya N, Yamaguchi A, Okaji Y, Tsuno NH, Kobata T, et al. Crucial Role of inhibitor of DNA binding/differentiation in the vascular endothelial growth factor induced activation and angiogenic processes of human endothelial cells. Journal of Immunology. 2004; 173:5801-5809.

8. Israel MA, Hernandez MC, Florio M, Andres-Barquin P, Mantani A, Carter JH, et al. Id gene expression as a key mediator of tumour cell biology. Cancer Research. 1999; 59:1726-1730.

9. Koch AE. Angiogenesis: implications for rheumatoid arthritis. Arthritis and Rheumatology. 1998; 41:951-962.

10. Lyden D, Young AZ, Zagzag D, Yan W, Gerald W, O'Reilly R, et al. Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts. Nature. 1999; 40:670-677.

11. Nishiyama K, Takaji K, Kataoka K, Kurihara Y, Yoshimura M, Kato A, et al. Id1 gene transfer confers angiogenic property on fully differentiated endothelial cells and contributes to therapeutic angiogenesis. Circulation. 2005; 112:2840–2850.

12. Ruzinova MB, Schoer RA, Gerald W, Egan JE, Pandolfi PP, Rafii S, et al. Effect of angiogenesis inhibition by Id loss and the contribution of bone marrow derived endothelial precuresor cells in spontaneous murine tumors. Cancer Cell. 2003; 4:277–289.

13. Kyei F. Knock-down of Id1 and Id3 proteins induces apoptosis in human colon carcinoma (HCT116) cells. Journal of Biology and Life Science. 2015; 6(1):194-208.

14. Hara E, Zebedee Z. Id proteins in cell cycle control and cellular senescence. Oncogene. 2001; 20:8317–8325.

15. Askew DS, Ashmun RA, Simmons BC, Clevaland JL. Constitutive c-myc expression in an IL-3-dependent myeloid cell line suppresses cell cycle arrest and accelerates apoptosis. Oncogene. 1991; 6:1915-1922.

16. Wagner AJ, Kokontis MJ, Hay N. Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21Cip1/Waf1. Genes and Development. 1994; 8:2817-2830.

17. Fong S, Debs RJ, Desprez PY. Id genes and proteins as promising targets in cancer therapy. Trends in Molecular Medicine. 2004; 10:387-92.

18. Tsuchiya T, Okaji Y, Tsuno NH, Sakurai D, Tsuchiya N, Kawai K, et al. Targeting Id1 and Id3 inhibit peritoneal metastasis of gastric cancer. Cancer Science. 2005; 96:784-790.

19. Hiebert SW, Packam G, Strom DK, Haffner R, Oren M, Zambetti G, et al. E2F-1: DP-1 induces p53 and overrides survival factors to trigger apoptosis. Molecular and Cellular Biology. 1995; 11:6864-68774.

20. Prabhu S, Ignatova A, Park ST, Sun XH. Regulation of expression of the cyclin dependent kinase inhibitor p21 by E2A and Id proteins. Molecular and Cellular Biology. 1997; 17:5888-5896.

21. Chen X, Ko JL, Jayaraman L, Prives C. P53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumour cells. Genes and Development. 1996; 11:2438-2451.

22. Florio M, Hernandez MC, Yang H, Shu HK, Cleveland JL, Israel MA. Id2 promotes apoptosis by a novel mechanism independent of dimerisation to basic helix-loop-helix factors. Molecular and Cellular Biology. 1998; 18:2371-2381.

23. Norton JD, Atherton GT. Coupling of cell growth control and apoptosis functions of ID proteins. Molecular and Cellular Biology. 1998; 18:2371-2381.

24. Yokota Y. Id and development. Oncogene. 2001; 20:8290-8298.

Article Metrics
86 Views 39 Downloads 125 Total

Year

Month

Related Search

By author names

Similar Articles