Targeting Apollon via siRNA Induces Apoptosis in Different Human Malignancies

Thumbnail Image

Date

2014

Journal Title

Journal ISSN

Volume Title

Type

Article

Publisher

IDOSI Publications

Series Info

Academic Journal of Cancer Research;7 (4): 233-238

Scientific Journal Rankings

Abstract

Apollon, also called Baculoviral IAP Repeat-Containing Protein 6 (BIRC6) or Baculoviral IAP Repeat-containing Ubiquitin Conjugating Enzyme (BRUCE), is an anti-apoptotic protein belonging to the IAP family, which consists of eight members. The genes of this family render cancer cells insensitive to apoptotic stimulation. The aim of the present study was to investigate and assess the role of small interference RNA (siRNA) in the regulation of Apollon gene expression in four different human cancerous cell lines; breast cancer (MCF-7), cervical cancer (HeLa), colon cancer (CaCo-2) and hepatocellular carcinoma (HepG-2). Lipofection was carried out to introduce the Apollon-specific siRNA into the cancerous cells and the Apollon expression levels were determined using RT-PCR. Trypan blue assay was conducted to assess the integrity of the cell membranes after being transfected. 3-(4, 5-dimethylthiazol-2-yl)-2-5- Diphenyl tetrazolium bromide (MTT) assay was also implemented to assess the cell viability through the mitochondrial reductase enzymes activity. The obtained results concluded that transfecting the malignant cells with Apollon-specific siRNA have led to the down regulation of Apollon expression compared to the control non-transfected cells. RNA interference targeting the anti-apoptotic genes such as Apollon could be considered as a promising approach and may help as a future therapeutic tool for many types of human cancers.

Description

MSA Google Scholar

Keywords

Apollon, SiRNA, Down-regulation, Apoptosis, MCF-7 HeLa CaCo-2 HepG-2

Citation

CAPMAS, 2011. Central Agency for Public required for normal placenta development and Mobilization and Statistics. Egyptian population mouse survival. Molecular and Cellular Biology, p y r a m i d . http://www.capmas.gov.eg/ 24(21): 9339-9350. show_cens.aspx. 24. Sabit, H., M.M.M. Ibrahim and N.S. Awad, 2013. 12. CIAUS, 2012. Central Intelligence Agency, US. The SiRNA-Mediated Down-Regulation of Livin world fact book. https://www.cia.gov/ Expression in Breast Cancer Cells. Academic Journal library/publications. of Cancer Research, 6(2): 69-73. 13. Fujiyama, S., M. Tanaka, S. Maeda, H. Ashihara, 25. Zhang, K., D. Chen, X. Wang, S. Zhang, J. Wang, R. Hirata and K. Tomita, 2002. Tumor markers in Y. Gao and B. Yan, 2011. RNA Interference Targeting early diagnosis, follow-up and management of Slug Increases Cholangiocarcinoma Cell Sensitivityto patients with hepatocellular carcinoma. Oncology, Cisplatin via up regulating PUMA. Int. J. Mol. Sci, 62(1): 57-63. 12: 385-400. 14. Marrero, J.A. and A.S. Lok, 2004. Newer markers 26. Palianopoulou, M., V. Papanikolaou, N. Stefanou and for hepatocellular carcinoma. Gastroenterology, A. Tsezou, 2011. The activation of leptin-mediated 127(5 Suppl 1): S113-S119. survivin is limited by the inducible suppressor SOCS15. Witjes, C.D., R.A. De Man, F.A. Eskens, 3 in MCF-7 cells. Exp Biol Med (Maywood), 236, 70-6. R.S. Dwarkasing, P.E. Zondervan, C. Verhoef and 27. Raj, D., T. Liu, G. Samadashwily, F. Li and D. J.N. Ijzermans, 2010. Hepatocellular carcinoma: the Grossman, 2008. Survivin repression by p53, Rb and significance of cirrhosis for treatment and prognosis E2F2 in normal human melanocytes. Carcinogenesis, retrospective study. Ned Tijdschr Geneeskd, 29(1): 194-201. 154: A1747. 28. Sabit, H., H.B. Al-Sharkawy, M. Al-Zaidy, M., Abdel16. Bauer, J.H. and S.L. Hefand, 2006. New tricks of an Baseer, N. S. Awad, W. N. Al-Mazny, N.S. Al-Saud, old molecule: lifespan regulation by p53. Aging Cell, M. M. Elsaadani, M.M.M. Ibrahim and O.A.M. Said, 5: 437-440. 2014. Small Interference RNA-Mediated Down 17. Kumar, V., A.K. Abbas, N. Fausto and J.C. Aster, Regulation of Survivin Expression in Breast and 2010. Robins and Cotran: pathologic basis of disease. Hepatocellular Carcinoma Cell Lines. World Journal Philadelphia: Saunders Elsevier, pp: 25-32. of Medical Sciences, 10(4): 421-426. 18. Ding, Z., G. Liu, B. Olsson and X. Sun, 2013. 29. Lopergolo, A., M. Pennati, P. Gandellini, N.I. Orlotti, Upregulation of the antiapoptotic factor Livins P. Poma and M.G. Daidone, 2009. Apollon gene contributes to cisplatin resistance in colon cancer silencing induces apoptosis in breast cancer cells cells. Tumor Biol., 34: 683-693. through p53 stabilization and caspase-3 activation. 19. Pepita, G.B., T. Avelina and P.T. Ricardo, 2009. British Journal of Cancer, 100: 739-746. Overcoming drug resistance by enhancing apoptosis 30. Lechler, P., T. Renkawitz, V. Campean, S. of tumor cells. Curr. Cancer Drug Targets, 9: 320-340. Balakrishnan, M. Tingart, J. Grifka and J. 20. Nachmias, B., Y. Ashhab and D. Ben-Yehuda, 2004. Schaumburger, 2011. The antiapoptotic gene survivin The inhibitor of apoptosis protein family (IAPs): an is highly expressed in human chondrosarcoma and emerging therapeutic target in cancer. Semin Cancer promotes drug resistance in chondrosarcoma cells in Biol., 14: 231-243. vitro. BMC Cancer, 11: 120. 21. Hauser, H.P., M. Bardroff, G. Pyrowolakis and 31. Chu, L., J. Gu, L. Sun, Q. Qian, C. Qian and X. Liu, S. Jentsch, 1998. A giant ubiquitin conjugating 2008. Oncolytic Adenovirus-Mediated Shrna Against enzyme related to IAP apoptosis inhibitors. Journal Apollon Inhibits Tumor Cell Growth And Enhances of Cell Biology, 141(6): 1415-1422. Antitumor Effect Of 5-Fluorouracil. Gene Ther., 22. Ren, J., M. Shi, R. Liu, Q. Yang, T. Johnson and 15: 484-94. W.C. Skarnes, 2005. The Birc6 (Bruce) gene regulates 32. Hendruschk, S., R. Wiedemuth, A. Aigner, K. Topfer, p53 and the mitochondrial pathway of apoptosis and M. Cartellieri, D. Martin, M. Kirsch, C. Ikonomidou, is essential for mouse embryonic development. G. Schackert and A. Temme, 2011. RNA Interference Proceedings of the National Academy of Sciences of Targeting Survivin Exerts Antitumoral Effects In Vitro the United States of America, 102(3): 565-570. And In Established Glioma Xenografts In Vivo. 23. Lotz, K., G. Pyrowolakis and S. Jentsch, 2004. Neuro Oncol., 13: 1074-89. BRUCE, a giant E2/E3 ubiquitin ligase and inhibitor of apoptosis protein of the trans-golgi network, is

Full Text link