Skip to main content
Top
Published in: Cancer Cell International 1/2012

Open Access 01-12-2012 | Primary research

2-Methoxyestradiol-bis-sulphamate refrains from inducing apoptosis and autophagy in a non-tumorigenic breast cell line

Authors: Michelle H Visagie, Anna M Joubert

Published in: Cancer Cell International | Issue 1/2012

Login to get access

Abstract

Background

Anticancer research resulted in the discovery of a promising antimitotic metabolite, 2-methoxyestradiol. 2-Methoxyestradiol-bis-sulphamate, a bis-sulphamoylated analogue exerts antiproliferative- and antimitotic activity. Investigating the anticancer potential of 2-methoxyestradiol-bis-sulphamate requires demonstrating the influence of 2-methoxyestradiol-bis-sulphamate on non-tumorigenic cells. This project focused on the in vitro effects of 2-methoxyestradiol-bis-sulphamate on the non-tumorigenic MCF-12A breast epithelial cell line.

Methods

The in vitro influence of 2-methoxyestradiol-bis-sulphamate was investigated on cell cycle progression, possible induction of apoptosis and autophagy and reactive oxygen species generation. Cell cycle progression was done using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Displaying effects on the mitochondrial membrane potential was achieved utilizing flow cytometry and the MitoCapture TM Mitochondrial apoptosis detection kit. Autophagy detection was done by means of flow cytometry and anti-LC3B conjugated to DyLight 488. Reactive oxygen species generation was conducted employing flow cytometry and 2,7-dichlorofluorescein diacetate and hydroethidine.

Results

This study demonstrated that 2-methoxyestradiol-bis-sulphamate did not affect cell cycle progression or reactive oxygen species in a statistically significant manner in the non-tumorigenic MCF-12A cell line. In addition, 2-methoxyestradiol-bis-sulphamate did not statistically significantly induce apoptosis or autophagy.

Conclusion

Reports indicate that 2-methoxyestradiol-bis-sulphamate induces apoptosis and autophagy in several tumorigenic cell lines. The anticancer ability of 2-methoxyestradiol-bis-sulphamate is due to its antimitotic activity. However, this study demonstrates the promising notion that 2-methoxyestradiol-bis-sulphamate does not affect the non-tumorigenic MCF-12A cells. This project contributes to the embedded scientific knowledge regarding the differential death mechanisms used by 2-methoxyestradiol-bis-sulphamate on tumorigenic and non-tumorigenic cell lines.
Appendix
Available only for authorised users
Literature
1.
go back to reference Stander A, Joubert F, Joubert A: Docking, synthesis, and in vitro evaluation of antimitotic estrone analogues. Chem Biol Drug Des. 2011, 77: 173-181. 10.1111/j.1747-0285.2010.01064.x.CrossRefPubMed Stander A, Joubert F, Joubert A: Docking, synthesis, and in vitro evaluation of antimitotic estrone analogues. Chem Biol Drug Des. 2011, 77: 173-181. 10.1111/j.1747-0285.2010.01064.x.CrossRefPubMed
2.
go back to reference Fotsis T, Zhang Y, Pepper MS, Aldercreutz H, Montesano R, Nawroth PP: Schweigerer: The endogenous oestrogen metabolite 2-methoxyestradiol inhibits angiogenesis and suppresses tumor growth. Nature. 1994, 368: 237-239. 10.1038/368237a0.CrossRefPubMed Fotsis T, Zhang Y, Pepper MS, Aldercreutz H, Montesano R, Nawroth PP: Schweigerer: The endogenous oestrogen metabolite 2-methoxyestradiol inhibits angiogenesis and suppresses tumor growth. Nature. 1994, 368: 237-239. 10.1038/368237a0.CrossRefPubMed
3.
go back to reference Risinger AL, Westbrook CD, Encinas M, Mülbaier M, Schultes CM, Wawro S, Lewis JD, Janssen B, Giles FJ, Moolberry SL: ELR5010444, a novel microtubule disruptor with multiple mechanisms of action. Pharmacol Exp Ther. 2011, 6: 652-660.CrossRef Risinger AL, Westbrook CD, Encinas M, Mülbaier M, Schultes CM, Wawro S, Lewis JD, Janssen B, Giles FJ, Moolberry SL: ELR5010444, a novel microtubule disruptor with multiple mechanisms of action. Pharmacol Exp Ther. 2011, 6: 652-660.CrossRef
4.
go back to reference Chander SK, Foster PA, Leese MP, Newman SP, Potter BVL, Purohit A, Reed MJ: In vivo inhibition of angiogenesis by sulphamoylated derivatives of 2-methoxyestradiol. Br J Cancer. 2007, 96: 1368-1376.PubMedCentralPubMed Chander SK, Foster PA, Leese MP, Newman SP, Potter BVL, Purohit A, Reed MJ: In vivo inhibition of angiogenesis by sulphamoylated derivatives of 2-methoxyestradiol. Br J Cancer. 2007, 96: 1368-1376.PubMedCentralPubMed
5.
go back to reference Visagie MH, Joubert AM: 2-Methoxyestradiol-bis-sulfamate induces apoptosis and autophagy in a tumorigenic breast epithelial cell line. Mol Cell Biochem. 2011, 357: 343-352. 10.1007/s11010-011-0905-3.CrossRefPubMed Visagie MH, Joubert AM: 2-Methoxyestradiol-bis-sulfamate induces apoptosis and autophagy in a tumorigenic breast epithelial cell line. Mol Cell Biochem. 2011, 357: 343-352. 10.1007/s11010-011-0905-3.CrossRefPubMed
6.
go back to reference Visagie MH, Joubert AM: The in vitro effects of 2-Methoxyestradiol-bis-sulphamate on cell numbers, membrane integrity and cell morphology, and the possible induction of apoptosis and autophagy in a non-tumorigenic breast epithelial cell line. Cell Mol Biol Lett. 2010, 15: 564-581. 10.2478/s11658-010-0030-4.CrossRefPubMed Visagie MH, Joubert AM: The in vitro effects of 2-Methoxyestradiol-bis-sulphamate on cell numbers, membrane integrity and cell morphology, and the possible induction of apoptosis and autophagy in a non-tumorigenic breast epithelial cell line. Cell Mol Biol Lett. 2010, 15: 564-581. 10.2478/s11658-010-0030-4.CrossRefPubMed
7.
go back to reference Ireson CR, Chander SK, Purohit A, Perera S, Newman SP, Parish D, Leese MP, Smith AC, Potter BVL, Reed MJ: Pharmacokinetics and efficacy of 2-methoxyoestradiol and 2-methoxyoestradiol-bis-sulphamate in vivo in rodents. Br J Cancer. 2004, 90: 932-937. 10.1038/sj.bjc.6601591.PubMedCentralCrossRefPubMed Ireson CR, Chander SK, Purohit A, Perera S, Newman SP, Parish D, Leese MP, Smith AC, Potter BVL, Reed MJ: Pharmacokinetics and efficacy of 2-methoxyoestradiol and 2-methoxyoestradiol-bis-sulphamate in vivo in rodents. Br J Cancer. 2004, 90: 932-937. 10.1038/sj.bjc.6601591.PubMedCentralCrossRefPubMed
8.
go back to reference Raobaikady B, Purohit A, Chander SK, Woo LWL, Leese MP, Potter BVL, Reed MJ: Inhibition of MCF-7 breast cancer cell proliferation and in vivo steroid sulphatase activity by 2-methoxyestradiol-bis-sulphamate. J Steroid Biochem Mol Biol. 2003, 84: 351-358. 10.1016/S0960-0760(03)00049-9.CrossRefPubMed Raobaikady B, Purohit A, Chander SK, Woo LWL, Leese MP, Potter BVL, Reed MJ: Inhibition of MCF-7 breast cancer cell proliferation and in vivo steroid sulphatase activity by 2-methoxyestradiol-bis-sulphamate. J Steroid Biochem Mol Biol. 2003, 84: 351-358. 10.1016/S0960-0760(03)00049-9.CrossRefPubMed
9.
go back to reference Suzuki RN, Newman SP, Purohit A, Leese MP, Potter BV, Reed MJ: Growth inhibition of multi-drug-resistant breast cancer cells by 2-methoxyoestradiol-bis-sulphamate and 2-ethyloestradiol-bis-sulphamate. J Steroid Biochem Mol Biol. 2003, 84: 269-278. 10.1016/S0960-0760(03)00035-9.CrossRefPubMed Suzuki RN, Newman SP, Purohit A, Leese MP, Potter BV, Reed MJ: Growth inhibition of multi-drug-resistant breast cancer cells by 2-methoxyoestradiol-bis-sulphamate and 2-ethyloestradiol-bis-sulphamate. J Steroid Biochem Mol Biol. 2003, 84: 269-278. 10.1016/S0960-0760(03)00035-9.CrossRefPubMed
10.
go back to reference Foster PA, Stengel C, Ali T, Leese MP, Potter BV, Reed MJ, Purohit A, Newman SP: A comparison of two orally bioavailable anti-cancer angents, IRC-110160 and STX140. Anticancer Res. 2008, 28: 1483-1491.PubMed Foster PA, Stengel C, Ali T, Leese MP, Potter BV, Reed MJ, Purohit A, Newman SP: A comparison of two orally bioavailable anti-cancer angents, IRC-110160 and STX140. Anticancer Res. 2008, 28: 1483-1491.PubMed
11.
go back to reference Raobaikady B, Reed MJ, Leese MP, Potter BVL, Purohit A: Inhibition of MDA-MB-231 cell cycle progression and cell proliferation by C-2-substituted oestradiol mono- and bis-3-O-sulphamates. Int J Cancer. 2005, 117: 150-159. 10.1002/ijc.21066.CrossRefPubMed Raobaikady B, Reed MJ, Leese MP, Potter BVL, Purohit A: Inhibition of MDA-MB-231 cell cycle progression and cell proliferation by C-2-substituted oestradiol mono- and bis-3-O-sulphamates. Int J Cancer. 2005, 117: 150-159. 10.1002/ijc.21066.CrossRefPubMed
12.
go back to reference Day JM, Foster PA, Tutill HJ, Newman SP, Ho YT, Leese MP, Potter BV, Reed MJ, Purohit A: BCRP expression does not result in resistance to STX140 in vivo, despite the increased expression of BCRP in A2780 cells in vitro after long-term STX140 exposure. Br J Cancer. 2009, 100: 476-486. 10.1038/sj.bjc.6604873.PubMedCentralCrossRefPubMed Day JM, Foster PA, Tutill HJ, Newman SP, Ho YT, Leese MP, Potter BV, Reed MJ, Purohit A: BCRP expression does not result in resistance to STX140 in vivo, despite the increased expression of BCRP in A2780 cells in vitro after long-term STX140 exposure. Br J Cancer. 2009, 100: 476-486. 10.1038/sj.bjc.6604873.PubMedCentralCrossRefPubMed
13.
go back to reference Newman SP, Leese MP, Purohit A, James DRC, Rennie CE, Potter BVL: Inhibition of in vitro angiogenesis by 2-Methoxy- and 2-ethyl-estrogen sulfamates. Int J Cancer. 2004, 109: 533-540. 10.1002/ijc.20045.CrossRefPubMed Newman SP, Leese MP, Purohit A, James DRC, Rennie CE, Potter BVL: Inhibition of in vitro angiogenesis by 2-Methoxy- and 2-ethyl-estrogen sulfamates. Int J Cancer. 2004, 109: 533-540. 10.1002/ijc.20045.CrossRefPubMed
14.
go back to reference Newman SP, Foster PA, Ho YT, Day JM, Raibaikady B, Kasprzyk PG, Leese MP, Potter BV, Reed MJ, Purohit A: The therapeutic potential of a series of bioavailable anti-angiogenic microtubule disruptors as therapy for hormone-independent prostate and breast cancers. Br J Cancer. 2007, 97: 1673-1682. 10.1038/sj.bjc.6604100.PubMedCentralCrossRefPubMed Newman SP, Foster PA, Ho YT, Day JM, Raibaikady B, Kasprzyk PG, Leese MP, Potter BV, Reed MJ, Purohit A: The therapeutic potential of a series of bioavailable anti-angiogenic microtubule disruptors as therapy for hormone-independent prostate and breast cancers. Br J Cancer. 2007, 97: 1673-1682. 10.1038/sj.bjc.6604100.PubMedCentralCrossRefPubMed
15.
go back to reference Wood L, Leese MP, Mouzakiti A, Purohit A, Potter BV, Reed MJ: Packham Gl: 2-MeOE2bisMATE induces caspase-dependent apoptosis in CAL51 breast cancer cells and overcomes resistance to TRAIL via cooperative activation of caspases. Apoptosis. 2004, 9: 323-332.CrossRefPubMed Wood L, Leese MP, Mouzakiti A, Purohit A, Potter BV, Reed MJ: Packham Gl: 2-MeOE2bisMATE induces caspase-dependent apoptosis in CAL51 breast cancer cells and overcomes resistance to TRAIL via cooperative activation of caspases. Apoptosis. 2004, 9: 323-332.CrossRefPubMed
16.
go back to reference Utsumi T, Leese MP, Chander SK, Gaukroger K, Purohit A, Newman SP, Potter BV, Reed MJ: The effects of 2-methoxyoestrogen sulphamates on the in vitro and in vivo proliferation of breast cancer cells. J Steroid Biochem Mol Biol. 2005, 94: 219-227. 10.1016/j.jsbmb.2005.01.022.CrossRefPubMed Utsumi T, Leese MP, Chander SK, Gaukroger K, Purohit A, Newman SP, Potter BV, Reed MJ: The effects of 2-methoxyoestrogen sulphamates on the in vitro and in vivo proliferation of breast cancer cells. J Steroid Biochem Mol Biol. 2005, 94: 219-227. 10.1016/j.jsbmb.2005.01.022.CrossRefPubMed
17.
go back to reference Leese LP, Jourdan FL, Gaokroger K, Mahon MF, Newman SP, Foster PA, Stengel C, Regis-Lydi S, Di Fiore A, De Simone G, Supuran CT, Purohit A, Reed MJ, Potter BVL: Structure-activity relationship of C-17 cyano-substituted estratriene as anticancer agents. J Med Chem. 2008, 51: 1295-1308. 10.1021/jm701319c.CrossRefPubMed Leese LP, Jourdan FL, Gaokroger K, Mahon MF, Newman SP, Foster PA, Stengel C, Regis-Lydi S, Di Fiore A, De Simone G, Supuran CT, Purohit A, Reed MJ, Potter BVL: Structure-activity relationship of C-17 cyano-substituted estratriene as anticancer agents. J Med Chem. 2008, 51: 1295-1308. 10.1021/jm701319c.CrossRefPubMed
18.
go back to reference Jourdan F, Leese MP, Dohle W, Hamel E, Ferrandism E, Newman SP, Purohit A, Rees MJ, Potter BVL: Synthesis, antitubulin, and antiproliferative SAR of analogues of 2-methoxyestradiol-3–17-O, O-bis-sulfamate. J Med Chem. 2010, 53: 2942-2951. 10.1021/jm9018806.CrossRefPubMed Jourdan F, Leese MP, Dohle W, Hamel E, Ferrandism E, Newman SP, Purohit A, Rees MJ, Potter BVL: Synthesis, antitubulin, and antiproliferative SAR of analogues of 2-methoxyestradiol-3–17-O, O-bis-sulfamate. J Med Chem. 2010, 53: 2942-2951. 10.1021/jm9018806.CrossRefPubMed
19.
go back to reference Newman SP, Ireson CR, Tutill HJ: The role of 17β-hydroxysteriod dehydrogenase in modulating the activity of 2-methoxyestradiol in breast cancer cells. Cancer Res. 2006, 66: 324-330. 10.1158/0008-5472.CAN-05-2391.CrossRefPubMed Newman SP, Ireson CR, Tutill HJ: The role of 17β-hydroxysteriod dehydrogenase in modulating the activity of 2-methoxyestradiol in breast cancer cells. Cancer Res. 2006, 66: 324-330. 10.1158/0008-5472.CAN-05-2391.CrossRefPubMed
20.
go back to reference Landes T, Martinou JC: Mitochondrialouter membrane permeabilization during apoptosis: the role of mitochondrial fission. Biochim Biophys Acta. 2011, 1813: 540-545. 10.1016/j.bbamcr.2011.01.021.CrossRefPubMed Landes T, Martinou JC: Mitochondrialouter membrane permeabilization during apoptosis: the role of mitochondrial fission. Biochim Biophys Acta. 2011, 1813: 540-545. 10.1016/j.bbamcr.2011.01.021.CrossRefPubMed
21.
go back to reference Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T: LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J. 2000, 19: 5720-5728. 10.1093/emboj/19.21.5720.PubMedCentralCrossRefPubMed Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T: LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J. 2000, 19: 5720-5728. 10.1093/emboj/19.21.5720.PubMedCentralCrossRefPubMed
22.
go back to reference Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, Tokuhisa T, Ohsumi Y, Yoshimori T: Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol. 2001, 152: 657-667. 10.1083/jcb.152.4.657.PubMedCentralCrossRefPubMed Mizushima N, Yamamoto A, Hatano M, Kobayashi Y, Kabeya Y, Suzuki K, Tokuhisa T, Ohsumi Y, Yoshimori T: Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J Cell Biol. 2001, 152: 657-667. 10.1083/jcb.152.4.657.PubMedCentralCrossRefPubMed
23.
go back to reference Mqoco T, Joubert A: Influence of estradiol analogue on cell growth, morphology and death in esophageal carcinoma cells. Biocell. 2010, 34: Mqoco T, Joubert A: Influence of estradiol analogue on cell growth, morphology and death in esophageal carcinoma cells. Biocell. 2010, 34:
24.
go back to reference Vorster CJJ, Joubert AM: In vitro effects of 2-methoxyestradiol-bis-sulphamate on the non-tumorigenic MCF-12A cell line. Cell Biochem Func. 2010, 28: 412-419. 10.1002/cbf.1671.CrossRef Vorster CJJ, Joubert AM: In vitro effects of 2-methoxyestradiol-bis-sulphamate on the non-tumorigenic MCF-12A cell line. Cell Biochem Func. 2010, 28: 412-419. 10.1002/cbf.1671.CrossRef
25.
go back to reference Visagie MH, Joubert AM: In vitro effects of 2-methoxyestradiol-bis-sulphamate on reactive oxygen species and possible apoptosis induction in a breast adenocarcinoma cell line. Cancer Cell Int. 2011, 11: 1-7. 10.1186/1475-2867-11-1.CrossRef Visagie MH, Joubert AM: In vitro effects of 2-methoxyestradiol-bis-sulphamate on reactive oxygen species and possible apoptosis induction in a breast adenocarcinoma cell line. Cancer Cell Int. 2011, 11: 1-7. 10.1186/1475-2867-11-1.CrossRef
26.
go back to reference Yang J, Wu LJ, Tashino SI, Onodera S, Ikejima T: reactive oxygen species and nitric oxide regulate mitochondrial-dependent apoptosis and autophagy in evodiamine-treated human cervix carcinoma HeLa cells. Free Radic Res. 2008, 42: 492-504. 10.1080/10715760802112791.CrossRefPubMed Yang J, Wu LJ, Tashino SI, Onodera S, Ikejima T: reactive oxygen species and nitric oxide regulate mitochondrial-dependent apoptosis and autophagy in evodiamine-treated human cervix carcinoma HeLa cells. Free Radic Res. 2008, 42: 492-504. 10.1080/10715760802112791.CrossRefPubMed
27.
go back to reference Circu ML, Aw TY: reactive oxygen species, cellular redox systems, and apoptosis. Free Rad Biol Med. 2010, 48: 749-762. 10.1016/j.freeradbiomed.2009.12.022.PubMedCentralCrossRefPubMed Circu ML, Aw TY: reactive oxygen species, cellular redox systems, and apoptosis. Free Rad Biol Med. 2010, 48: 749-762. 10.1016/j.freeradbiomed.2009.12.022.PubMedCentralCrossRefPubMed
28.
go back to reference Shrotriya S, Deep G, Gu M, Kaur M, Jain AK, Inturi S, Agarwal R, Agarwal C: generation of reactive oxygen species by grape seed extract causes irreparable DNA damage leading to G2/M arrest and apoptosis selectively in head and neck sqamous cell carcinoma cells. Carcimogenesis. 2012, in press Shrotriya S, Deep G, Gu M, Kaur M, Jain AK, Inturi S, Agarwal R, Agarwal C: generation of reactive oxygen species by grape seed extract causes irreparable DNA damage leading to G2/M arrest and apoptosis selectively in head and neck sqamous cell carcinoma cells. Carcimogenesis. 2012, in press
29.
go back to reference Azad MB, Chen Y, Gibson B: Regulation of autophagy by reactive oxygen species (ROS): implications for cancer progression and cancer. Antioxid Redox Signal. 2009, 11 (4): 1-14.CrossRef Azad MB, Chen Y, Gibson B: Regulation of autophagy by reactive oxygen species (ROS): implications for cancer progression and cancer. Antioxid Redox Signal. 2009, 11 (4): 1-14.CrossRef
Metadata
Title
2-Methoxyestradiol-bis-sulphamate refrains from inducing apoptosis and autophagy in a non-tumorigenic breast cell line
Authors
Michelle H Visagie
Anna M Joubert
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2012
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/1475-2867-12-37

Other articles of this Issue 1/2012

Cancer Cell International 1/2012 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine