Inhibitory Effects of Curcumin and 2-Methoxyestradiaul on Anaphase Promoting Complex in Hela Cancer Cells

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

2 Professor, Department of Pharmaceutics, School of Pharmacy AND Pharmaceutical Science Research Center, Tehran University of Medical Sciences, Tehran, Iran

3 Associate Professor, Department of Biotechnology, National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

4 Assistant Professor, Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

5 Assistant Professor, Department of Life Sciences, Barcelona Supercomputing Center (BSC), Barcelona, Spain

Abstract

Background: Anaphase promoting complex (APC) plays a critical role in cell division and mitotic exit. This protein complex may have a pivotal role in the cell cycle control affecting pathological conditions such as cancer. APC is recommended as the target of many anti-cancer agents due to its importance in cancer pathogenesis. This study aimed to evaluate the inhibitory effects of curcumin and 2-methoxyestradiaul on APC. Methods: Hela cells were treated with various concentrations of curcumin and 2-methoxyestradiaul, and their cytotoxic effects were investigated after 24, 48, 72 and 96 hours by MTT assay. Expression of securin, cyclin B and the APC substrates were investigated using immune-blotting. Finally, cell cycle analysis was performed using DRAQ5 staining and flowcytometry.Findings: Respectively, 20 and 30 percent of cell-death after 24-hour treatment with curcumin and 2-methoxyestradiaul was seen. Also, securin and cyclin levels were raised after 24 hours of treatment. Furthermore, the levels of the substrates (securin and cyclin B) increased 48 hours after treatment. Results of fluorescence-activated cell sorting (FACS) analysis showed that treated cells were arrested in G2 phase of cell cycle which revealed cell arrest in G2 phase of cell cycle. 2-methoxyestradiaul showed a better APC inhibition effect than curcumin (P = 0.02). Conclusions: Our results revealed that APC is a suitable target for cancer treatment and curcumin and 2-methoxyestradiaul have inhibitory effects on the activity of this complex.

Keywords

References

  1. Siegel R, Ward E, Brawley O, Jemal A. Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin 2011; 61(4): 212-36.
  2. Ministry of Health and Medical Education, Department of Health. Iranian annual cancer registration report 2003. Tehran, Iran: Kelke-Dirin; 2005. [In Persian].
  3. Tessema M, Lehmann U, Kreipe H. Cell cycle and no end. Virchows Arch 2004; 444(4): 313-23.
  4. Ingolia N. Cell cycle: bistability is needed for robust cycling. Curr Biol 2005; 15(23): R961-R963.
  5. Nakayama KI, Nakayama K. Ubiquitin ligases: cell-cycle control and cancer. Nat Rev Cancer 2006; 6(5): 369-81.
  6. Deshaies RJ. SCF and Cullin/Ring H2-based ubiquitin ligases. Annu Rev Cell Dev Biol 1999; 15: 435-67.
  7. Irniger S, Nasmyth K. The anaphase-promoting complex is required in G1 arrested yeast cells to inhibit B-type cyclin accumulation and to prevent uncontrolled entry into S-phase. J Cell Sci 1997; 110 ( Pt 13): 1523-31.
  8. Narbonne-Reveau K, Senger S, Pal M, Herr A, Richardson HE, Asano M, et al. APC/CFzr/Cdh1 promotes cell cycle progression during the Drosophila endocycle. Development 2008; 135(8): 1451-61.
  9. Turnell AS, Stewart GS, Grand RJ, Rookes SM, Martin A, Yamano H, et al. The APC/C and CBP/p300 cooperate to regulate transcription and cell-cycle progression. Nature 2005; 438(7068): 690-5.
  10. Herzog F, Primorac I, Dube P, Lenart P, Sander B, Mechtler K, et al. Structure of the anaphase-promoting complex/cyclosome interacting with a mitotic checkpoint complex. Science 2009; 323(5920): 1477-81.
  11. Wickliffe K, Williamson A, Jin L, Rape M. The multiple layers of ubiquitin-dependent cell cycle control. Chem Rev 2009; 109(4): 1537-48.
  12. Lehman NL, Tibshirani R, Hsu JY, Natkunam Y, Harris BT, West RB, et al. Oncogenic regulators and substrates of the anaphase promoting complex/cyclosome are frequently overexpressed in malignant tumors. Am J Pathol 2007; 170(5): 1793-805.
  13. Mondal G, Sengupta S, Panda CK, Gollin SM, Saunders WS, Roychoudhury S. Overexpression of Cdc20 leads to impairment of the spindle assembly checkpoint and aneuploidization in oral cancer. Carcinogenesis 2007; 28(1): 81-92.
  14. Gutgemann I, Lehman NL, Jackson PK, Longacre TA. Emi1 protein accumulation implicates misregulation of the anaphase promoting complex/cyclosome pathway in ovarian clear cell carcinoma. Mod Pathol 2008; 21(4): 445-54.
  15. Kang Y, Kim JH, Lee TH, Kim TS, Jung WH, Chung HC, et al. Expression of anaphase-promoting complex7 in fibroadenomas and phyllodes tumors of breast. Hum Pathol 2009; 40(1): 98-107.
  16. Fujita T, Liu W, Doihara H, Date H, Wan Y. Dissection of the APCCdh1-Skp2 cascade in breast cancer. Clin Cancer Res 2008; 14(7): 1966-75.
  17. Wang Q, Moyret-Lalle C, Couzon F, Surbiguet-Clippe C, Saurin JC, Lorca T, et al. Alterations of anaphase-promoting complex genes in human colon cancer cells. Oncogene 2003; 22(10): 1486-90.
  18. Kim JM, Sohn HY, Yoon SY, Oh JH, Yang JO, Kim JH, et al. Identification of gastric cancer-related genes using a cDNA microarray containing novel expressed sequence tags expressed in gastric cancer cells. Clin Cancer Res 2005; 11(2 Pt 1): 473-82.
  19. Montero A, Fossella F, Hortobagyi G, Valero V. Docetaxel for treatment of solid tumours: a systematic review of clinical data. Lancet Oncol 2005; 6(4): 229-39.
  20. Manchado E, Guillamot M, Malumbres M. Killing cells by targeting mitosis. Cell Death Differ 2012; 19(3): 369-77.
  21. Jordan MA, Wilson L. Microtubules as a target for anticancer drugs. Nat Rev Cancer 2004; 4(4): 253-65.
  22. Mena AL, Lam EW, Chatterjee S. Sustained spindle-assembly checkpoint response requires de novo transcription and translation of cyclin B1. PLoS One 2010; 5(9).
  23. Perez EA. Microtubule inhibitors: Differentiating tubulin-inhibiting agents based on mechanisms of action, clinical activity, and resistance. Mol Cancer Ther 2009; 8(8): 2086-95.
  24. Gottesman MM. Mechanisms of cancer drug resistance. Annu Rev Med 2002; 53: 615-27.
  25. Zeng X, King RW. An APC/C inhibitor stabilizes cyclin B1 by prematurely terminating ubiquitination. Nat Chem Biol 2012; 8(4): 383-92.
  26. Hatcher H, Planalp R, Cho J, Torti FM, Torti SV. Curcumin: from ancient medicine to current clinical trials. Cell Mol Life Sci 2008; 65(11): 1631-52.
  27. Goel A, Kunnumakkara AB, Aggarwal BB. Curcumin as "Curecumin": from kitchen to clinic. Biochem Pharmacol 2008; 75(4): 787-809.
  28. Sa G, Das T. Anti cancer effects of curcumin: cycle of life and death. Cell Div 2008; 3: 14.
  29. Lee SJ, Langhans SA. Anaphase-promoting complex/cyclosome protein Cdc27 is a target for curcumin-induced cell cycle arrest and apoptosis. BMC Cancer 2012; 12: 44.
  30. Bhati R, Gokmen-Polar Y, Sledge GW, Jr., Fan C, Nakshatri H, Ketelsen D, et al. 2-methoxyestradiol inhibits the anaphase-promoting complex and protein translation in human breast cancer cells. Cancer Res 2007; 67(2): 702-8.
  31. Zhao J, Zhao Y, Zhang Y, Chen W. Anti-tumor effect of curcumin on human cervical carcinoma HeLa cells in vitro and in vivo. Chin J Cancer Res 2007; 19(1): 32-6.
  32. Salama SA, Kamel MW, Botting S, Salih SM, Borahay MA, Hamed AA, et al. Catechol-o-methyltransferase expression and 2-methoxyestradiol affect microtubule dynamics and modify steroid receptor signaling in leiomyoma cells. PLoS One 2009; 4(10): e7356.
  33. Wimbauer F, Yang C, Shogren KL, Zhang M, Goyal R, Riester SM, et al. Regulation of interferon pathway in 2-methoxyestradiol-treated osteosarcoma cells. BMC Cancer 2012; 12: 93.
  34. Plum SM, Park EJ, Strawn SJ, Moore EG, Sidor CF, Fogler WE. Disease modifying and antiangiogenic activity of 2-methoxyestradiol in a murine model of rheumatoid arthritis. BMC Musculoskelet Disord 2009; 10: 46.
  35. Xia G, Chen B, Ding J, Gao C, Lu H, Shao Z, et al. Effect of magnetic Fe3O4 nanoparticles with 2-methoxyestradiol on the cell-cycle progression and apoptosis of myelodysplastic syndrome cells. Int J Nanomedicine 2011; 6: 1921-7.
Journal of Isfahan Medical School
Volume 31, Issue 228 - Serial Number 228
January and February 2013
Pages 255-264

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History

  • Receive Date: 16 March 2013
  • Revise Date: 26 May 2024
  • Accept Date: 06 April 2022

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