Downregulation of KDR expression induces apoptosis in breast cancer cells

• Autorzy: Zhang X. , Ge Y.-L. , Zhang S.-P. , Yan P. , Tian R.-H.
• Języki publikacji: EN

Abstrakty

EN

Angiogenesis plays a crucial role in the growth, invasion and metastasis of breast cancer. Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are the key regulators of tumor angiogenesis. VEGFR-2, known as the kinase insert domain receptor (KDR), is a key receptor involved in malignant angiogenesis. We previously showed that knocking down KDR with short interference RNA (KDR-siRNA) markedly decreased KDR expression and suppressed tumor growth in a xenograft model. However, the mechanisms underlying the anti-cancer effects of KDR-siRNA are not clearly understood. This study aimed to elucidate the molecular mechanisms that induce apoptosis in human breast cancer MCF-7 cells after transfection with KDR-siRNA. We studied the effects of KDR-siRNA on proliferation, apoptosis, antiapoptotic and pro-apoptotic proteins, mitochondrial membrane permeability, cytochrome c release and caspase-3 activity. The results indicated that KDR-siRNA treatment significantly inhibited the proliferation and induced the apoptosis of MCF-7 cells, reduced the levels of the anti-apoptotic proteins, Bcl-2 and Bcl-xl, and increased the level of the pro-apoptotic protein Bax, resulting in a decreased Bcl-2/Bax ratio. KDR-siRNA also enhanced the mitochondrial membrane permeability, induced cytochrome c release from the mitochondria, upregulated apoptotic protease-activating factor-1 (Apaf-1), cleaved caspase-3, and increased caspase-3 activity in MCF-7 cells. Furthermore, KDR-siRNA-induced apoptosis in MCF-7 cells was blocked by the caspase inhibitor Z-VAD-FMK, suggesting a role of caspase activation in the induction of apoptosis. These results indicate that the Bcl-2 family proteins and caspase-related mitochondrial pathways are primarily involved in KDR-siRNAinduced apoptosis in MCF-7 cells and that KDR might be a potential therapeutic target for human breast cancer treatments.

Słowa kluczowe

EN

angiogenesis; kinase domain receptor; apoptosis; breast cancer cell; cancer cell; bovine serum albumin; vascular endothelial growth factor; vascular endothelial growth factor receptor

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2014
• Tom: 19
• Numer: 4
• Opis fizyczny: p.527-541,fig.,ref.

Twórcy

autor

Zhang X.

• Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, 38 Dengzhou Road, Qingdao, China, 266021

autor

Ge Y.-L.

• Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, 38 Dengzhou Road, Qingdao, China, 266021

autor

Zhang S.-P.

• Department of Obstetrics, Qingdao Municipal Hospital, 5 Donghai middle Road, Qingdao, China, 266011

autor

Yan P.

• Department of Oral, Zichuan Hospital, 591 Zicheng Road, Zibo, China, 255100

autor

Tian R.-H.

• Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, 38 Dengzhou Road, Qingdao, China, 266021

Bibliografia

• 1.Jemal, A., Bray, F., Center, M.M., Ferlay, J., Ward, E. and Forman, D. Global cancer statistics. CA Cancer J. Clin. 61 (2011) 69–90.
• 2. Linos, E., Spanos, D. , Rosner, B.A., Linos, K., Hesketh, T., Qu, J.D., Gao, Y.T., Zheng, W. and Colditz, G.A. Effects of reproductive and demographic changes on breast cancer incidence in China: a modeling analysis. J. Natl. Cancer Inst. 100 (2008) 1352–1360.
• 3. Dai, M., Ren, J.S., Li, N., Li, Q., Yang, L. and Chen, Y.H. Estimation and prediction on cancer related incidence and mortality in China, 2008. Zhonghua Liu. Xing. Bing. Xue. Za. Zhi. 33 (2012) 57–61.
• 4. Hyder, S.M. Sex-steroid regulation of vascular endothelial growth factor in breast cancer. Endocr. Relat. Cancer. 13 (2006) 667–687.
• 5. Takekoshi, K., Isobe, K., Yashiro, T., Hara, H., Ishii, K., Kawakami, Y., Nakai, T. and Okuda, Y. Expression of vascular endothelial growth factor (VEGF) and its cognate receptors in human pheochromocytomas. Life Sci. 74 (2004) 863–871.
• 6. Roskoski, R. Jr. Vascular endothelial growth factor (VEGF) signaling in tumor progression. Crit. Rev. Oncol. Hematol. 62 (2007) 179–213.
• 7. Vieira, J.M., Santos, S.C., Espadinha, C., Correia, I., Vag, T., Casalou, C., Cavaco, B.M., Catarino, A.L., Dias, S. and Leite, V. Expression of vascular endothelial growth factor (VEGF) and its receptors in thyroid carcinomas of follicular origin: a potential autocrine loop. Eur. J. Endocrinol. 153 (2005) 701–709.
• 8. Guo, S., Colbert, L.S., Fuller, M., Zhang, Y. and Gonzalez-Perez, R.R. Vascular endothelial growth factor receptor-2 in breast cancer. Biochim. Biophys. Acta 1806 (2010) 108–121.
• 9. Takahashi, Y., Kitadai, Y., Bucana, C.D., Cleary, K.R. and Ellis, L.M. Expression of vascular endothelial growth factor and its receptor, KDR, correlates with vascularity, metastasis and proliferation of human colon cancer. Cancer Res. 55 (1995) 3964–3968.
• 10. Ryden, L., Linderholm, B., Nielsen, N.H., Emdin, S., Jonsson, P.E. and Landberg, G. Tumor specific VEGF-A and VEGFR2/KDR protein are coexpressed in breast cancer. Breast Cancer Res. Treat. 82 (2003) 147–154.
• 11. Spratlin, J. Ramucirumab (IMC-1121B): Monoclonal antibody inhibition of vascular endothelial growth factor receptor-2. Curr. Oncol. Rep. 13 (2011) 97–102.
• 12. Hammond, S.M., Bexnstein, E., Beach. D. and Hannon, G.J. An RNAdirected nuclease mediates post-transcriptional gene silencing in Dxosophila cells. Nature 404 (2000) 293.
• 13. Ge, Y.L., Zhang, J.Y., Zhang, X., Hou, L., Li, Q. and Xue, M.L. Chemically modified siRNA directed against the KDR gene inhibits the proliferation of breast cancer cells. Mol. Med. Rep. 2 (2009) 121–127.
• 14. Kim, E.J., Park, S.Y., Lee, J.Y. and Park, J.H. Fucoidan present in brown algae induces apoptosis of human colon cancer cells. BMC Gastroenterol. 10 (2010) 96.
• 15. Minamino, M., Sakaguchi, I., Naka, T., Iked, N., Katoa, Y., Tomiyasu, I., Yano, I. and Kobayashi, K. Bacterial ceramides and sphingophospholipids induce apoptosis of human leukaemic cells. Microbiology 149 (2003) 2071– 2081.
• 16. Chen, Q., Li, L., Tu, Y., Zheng, L.L., Liu, W., Zuo, X.Y., He, Y.M., Zhang, S.Y., Zhu, W., Cao. J.P., Cui, F.M. and Hou, J. MiR-34a regulates apoptosis in liver cells by targeting the KLF4 gene. Cell. Mol. Biol. Lett. 19 (2014) 52–64.
• 17. Westphal, S. and Kalthoff, H. Apoptosis: targets in pancreatic cancer. Mol. Cancer 2 (2003) 6.
• 18. Kuo, P.C., Liu, H.F. and Chao, J.I. Survivin and p53 modulate quercetininduced cell growth inhibition and apoptosis in human lung carcinoma cells. J. Biol. Chem. 279 (2004) 55875–55885.
• 19. Katiyar, S.K., Roy, A.M. and Baliga, M.S. Silymarin induces apoptosis primarily through a p53-dependent pathway involving Bcl-2/Bax, cytochrome c release and caspase activation. Mol. Cancer Ther. 4 (2005) 207–216.
• 20. Jin, Z. and El-Deiry, W.S. Overview of cell death signaling pathways. Cancer Biol. Ther. 4 (2005) 139–163.
• 21. Ghobrial, I.M., Witzig, T.E. and Adjei, A.A. Targeting apoptosis pathways in cancer therapy. CA Cancer J. Clin. 55 (2005) 178–194.
• 22. Jagani, H., Rao, J.V., Palanimuthu, V.R., Hariharapura, R.C. and Gang, S. A nanoformulation of siRNA and its role in cancer therapy: in vitro and in vivo evaluation. Cell. Mol. Biol. Lett. 18 (2013) 120–136.
• 23. Reed, J.C. Regulation of apoptosis by Bcl-2 family proteins and its role in cancer and chemoresistance. Curr. Opin. Oncol. 7 (1995) 541–546.
• 24. Adams, J.M. and Cory, S. Bcl-2-regulated apoptosis: mechanism and therapeutic potential. Curr. Opin. Immunol. 19 (2007) 488–496.
• 25. Kumari, A. and Kakkar, P. Lupeol prevents acetaminophen-induced in vivo hepatotoxicity by altering the Bax/Bcl-2- and oxidative stress-mediated mitochondrial signaling cascade. Life Sci. 90 (2012) 561–570.
• 26. Chen, Y.C., Shen, S.C., Lee, W.R., Hsu, F.L., Lin, H.Y., Ko, C.H. and Tseng, S.W. Emodin induces apoptosis in human promyelokukemic HL-60 cells accompanied by activation of caspase-3 cascade but independent of reactive oxygen species production. Biochem. Pharmacol. 64 (2002) 1713– 1724.

Identyfikatory

DOI

10.2478/s11658-014-0210-8