Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Cancer Cell International
Published in: Cancer Cell International 1/2016

Open Access 01-12-2016 | Primary research

Autophagy induced by a sulphamoylated estrone analogue contributes to its cytotoxic effect on breast cancer cells

Authors: Marcel Verwey, Elsie M. Nolte, Anna M. Joubert, Anne E. Theron

Published in: Cancer Cell International | Issue 1/2016

Login to get access

Abstract

Background

Autophagy can either be protective and confer survival to stressed cells, or it can contribute to cell death. The antimitotic drug 2-ethyl-3-O-sulpamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) is an in silico-designed 17-β-estradiol analogue that induces both autophagy and apoptosis in cancer cells. The aim of the study was to determine the role of autophagy in ESE-15-ol-exposed human adenocarcinoma breast cancer cells; knowledge that will contribute to future clinical applications of this novel antimitotic compound. By inhibiting autophagy and determining the cytotoxic effects of ESE-15-ol-exposure, deductions could be made as to whether the process may confer resistance to the drug, or alternatively, contribute to the cell death process.

Methods and results

Spectophometrical analysis via crystal violet staining was used to perform cytotoxicity studies. Morphology studies were done using microscopic techniques namely polarization-optical transmitted light differential interference light microscopy, fluorescent microscopy using monodansylcadaverine staining and transmission electron microscopy. Flow cytometry was used to quantify the autophagy inhibition and assess cell viability. Results obtained indicated that 3-methyladenine inhibited autophagy and increased cell survival in both MCF-7 and MDA-MB-231 cell lines.

Conclusion

This in vitro study inferred that autophagy inhibition with 3-methyladenine does not confer increased effectiveness of ESE-15-ol in inducing cell death. Thus it may be concluded that the autophagic process induced by ESE-15-ol exposure in MCF-7 and MDA-MB-231 cells plays a more significant role in cell death than conferring survival.
Literature
1.
Lewis J, Thomas T, Pestell R. Differential effects of 16α-hydroxyestrone and 2-methoxyestradiol on cyclin d1 involving the transcription factor atf-2 in mcf-7 breast cancer cells. J Mol Endocrinol. 2005;34:94–105.CrossRef
2.
Fishman J, Bradlow H, Gallagher T. Oxidative metabolism of estradiol. J Biol Chem. 1960;235:3104–7.PubMed
3.
Li L, Bu S, Bäckström T, Landström M, Ulmsten U, Fu X. Induction of apoptosis and G2/m arrest by 2-methoxyestradiol in human cervical cancer HeLaS3 cells. Anticancer Res. 2004;24:873–80.PubMed
4.
Leese MP, Leblond B, Smith A, Newman SP, Di Fiore A, De Simone G, Supuran CT, Purohit A, Reed MJ, Potter BVL. 2-substituted estradiol bis-sulfamates, multitargeted antitumor agents: synthesis, in vitro sar, protein crystallography, and in vivo activity. J Med Chem. 2006;49:7683–96.CrossRefPubMed
5.
Stander B, Marais S, Vorster C, Joubert A. In vitro effects of 2-methoxyestradiol on morphology, cell cycle progression, cell death and gene expression changes in tumorigenic mcf-7 breast epithelial cell line. J Steroid Biochem Mol Biol. 2010;119:149–60.CrossRefPubMed
6.
7.
Zhou N, Zhu X, Zhou J, Li M, Zhang X, Huang P, Jiang W. 2-Methoxyestradiol induces cell cycle arrest and apoptosis of nasopharyngeal carcinoma cells. Acta Pharmacol Sin. 2004;25:1515–20.PubMed
8.
Potapenko O, Zemskova M, Verin A, Zemskov E. Microtubule disruptor 2-methoxyestradiol has pro-inflammatory effect on human pulmonary endothelial cells. In: Inflammation and vascular permeability. New York: American Thoracic Society; 2014. pp A3268.
9.
Stander BA, Joubert F, Tu C, Joubert AM. In vitro evaluation of ese-15-ol, an estradiol analogue with nanomolar antimitotic and carbonic anhydrase inhibitory activity. PLoS ONE. 2012;7:e52205.CrossRefPubMedPubMedCentral
10.
Gascoigne K, Taylor S. How do anti-mitotic drugs kill cancer cells? J Cell Sci. 2009;122:2579–85.CrossRefPubMed
11.
Theron A, Prudent R, Nolte E, Bout I, Punchoo R, Marais S, du Toit P, Hlophe Y, Papendorp D, Lafanechère L, et al. Novel in silico-designed estradiol analogues are cytotoxic to a multidrug-resistant cell line at nanomolar concentrations. Cancer Chemother Pharmacol. 2014;75:431–7.CrossRefPubMed
12.
Elger W, Schwarz S, Hedden A, Reddersen G, Schneider B. Sulfamates of various estrogens are prodrugs with increased systemic and reduced hepatic estrogenicity at oral application. J Steroid Biochem Mol Biol. 1995;55:395–403.CrossRefPubMed
13.
Winum J, Scozzafava A, Montero J, Supuran C. Sulfamates and their therapeutic potential. Medic Res Rev. 2005;25:186–228.CrossRef
14.
Genis C, Sippel K, Case N, Cao W, Avvaru B, Tartaglia L, Govindasamy L, Tu C, Agbandje-McKenna M, Silverman D, et al. Design of a carbonic anhydrase ix active-site mimic to screen inhibitors for possible anticancer properties. Biochemistry. 2009;48:1322–31.CrossRefPubMedPubMedCentral
15.
Fukumura D, Jain R. Tumor microenvironment abnormalities: causes, consequences, and strategies to normalize. J Cell Biochem. 2007;101:937–49.CrossRefPubMed
16.
17.
Johnson B, Ober W, Garrison C, Silverthorn A. Human physiology: an integrated approach. 5th ed. New York: Pearson; 2010.
18.
Vermes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death. J Immunol Methods. 2000;243:167–90.CrossRefPubMed
19.
Ravikumar B, Berger Z, Vacher C, O’Kane C, Rubinsztein D. Rapamycin pre-treatment protects against apoptosis. Hum Mol Genet. 2006;15:1209–16.CrossRefPubMed
20.
Meléndez A, Levine B. Autophagy in C. Elegans. Beijing: Worm Book; 2009.
21.
22.
Bandyopadhyay U, Kaushik S, Varticovski L, Cuervo A. The chaperone-mediated autophagy receptor organizes in dynamic protein complexes at the lysosomal membrane. Mol Cell Biol. 2008;28:5754–63.CrossRef
23.
Wu YT, Tan H. Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class i and iii phosphoinositide 3-kinase. J Biol Chem. 2010;285:10850–61.CrossRefPubMedPubMedCentral
24.
Tran A, Ramalinga M, Kedir H, Clarke R, Kumar D. Autophagy inhibitor 3-methyladenine potentiates apoptosis induced by dietary tocotrienols in breast cancer cells. Euro J Nutr. 2014;54:265–72.CrossRef
25.
Ito S, Koshikawa N, Mochizuki S, Takenaga K. 3-Methyladenine suppresses cell migration and invasion of ht1080 fibrosarcoma cells through inhibiting phosphoinositide 3-kinases independently of autophagy inhibition. Int J Oncol. 2007;31:261–8.PubMed
26.
Li J, Hou N, Faried A, Tsutsumi S, Kuwano H. Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model. Euro J Cancer. 2010;46:1900–9.CrossRef
27.
Xie B, Zhao H, Yao S, Zhuo D, Jin B, Lv D, Ai Z. Autophagy inhibition enhances etoposide-induced cell death in human hepatoma g2 cells. Inter J Mol Med. 2011;27:599–606.
28.
Bonet-Ponce L, Saez-Atienzar S, Casa C, Sancho-Pelluz J, Barcia JM, Martinez-Gil N, Nava E, Jordan J, Romero FJ, Galindo MF. Rotenone induces the formation of 4-hydroxynonenal aggresomes: role of ROS-mediated tubulin hyperacetylation and autophagic flux disruption. Mol Neurobiol. 2015;53:1–15.
29.
30.
Gillies R, Didier N. Determination of cell number in monolayer cultures. Anal Biochem. 1986;159:109–13.CrossRefPubMed
31.
Krishan A. Rapid flow cytofluorometris analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975;66:188–93.CrossRefPubMed
32.
33.
34.
Niemann A, Takatsuki A. The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe. J Histochem Cytochem. 2000;48:251–8.CrossRefPubMed
35.
Vasquez RJ, Howell B, Yvon AM, Wadsworth P, Cassimeris L. Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro. Mol Biol Cell. 1997;8:973–85.CrossRefPubMedPubMedCentral
36.
Theron J, Nolte E, Lafanechère L, Joubert AM. Molecular crosstalk between apoptosis and autophagy induced by a novel 2-methoxyestradiol analogue in cervical adenocarcinoma cells. Cancer Cell Int. 2013;13:87.CrossRefPubMedPubMedCentral
37.
Visagie M, Joubert A. The in vitro effects of 2-methoxyestradiol-bis-sulfphamate on cell number, mebrane integrity and cell morphology, and the possible induction of apoptosis and autophagy in a non-tumorogenicbreast epithelial cell line. Cell Mol Biol Lett. 2010;15:564–81.CrossRefPubMed
38.
Stander BA, Joubert F, Joubert A. Docking synthesis and in vitro evaluation of antimitotic estrone analogs. Chem Biol Drug Design. 2011;77:173–81.CrossRef
39.
Wolmarans E, Mqoco T, Stander A, Nkandeu S, Sippel K, McKenna R, Joubert A. Novel estradiol analogue induces apoptosis and autophagy in esophageal carcinoma cells. Cell Mol Biol Lett. 2014;19:98.CrossRefPubMed
40.
Stander BA, Joubert F, Tu C, Sippel KH, McKenna R, Joubert A. Signaling pathways of ESE-16, an antimitotic and anticarbonic anhydrase estradiol analog, in breast cancer cells. PLoS ONE. 2013;8:e53853.CrossRefPubMedPubMedCentral
41.
Visagie M, Mqoco T, Joubert A. Sulphamoylated estradiol analogue induces antiproliferative activity and apoptosis in breast cell lines. Cell Mol Biol Lett. 2012;17:549.CrossRefPubMed
42.
Visagie MH, Stander BA, Birkholtz L, Joubert AM. Short communication: effects of a 17-beta estradiol analogue on gene expression and morphology in a breast epithelial adenocarcinoma cell line: a potential antiproliferative agent. Biomed Res. 2013;24:525–30.
43.
Yang X, Yu D-D, Yan F, Jing Y-Y, Han Z-P, Sun K, Liang L, Hou J, Wei L-X. The role of autophagy induced by tumor microenvironment in different cells and stages of cancer. Cell Biosci. 2015;5:14.CrossRefPubMedPubMedCentral
44.
Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, Adachi H, Adams CM, Adams PD, Adeli K, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 2016;12:1–222.CrossRefPubMed
45.
Menzies FM, Moreau K, Puri C, Renna M, Rubinsztein DC. Measurement of autophagic activity in mammalian cells. In: Current protocols cell biology. New York: Wiley; 2001.
46.
Desai BN, Myers BR, Schreiber SL. Fkbp12-rapamycin-associated protein associates with mitochondria and senses osmotic stress via mitochondrial dysfunction. Proc Natl Acad Sci USA. 2002;99:4319–24.CrossRefPubMedPubMedCentral
Metadata
Title
Autophagy induced by a sulphamoylated estrone analogue contributes to its cytotoxic effect on breast cancer cells
Authors
Marcel Verwey
Elsie M. Nolte
Anna M. Joubert
Anne E. Theron
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2016
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-016-0367-5

Other articles of this Issue 1/2016

Cancer Cell International 1/2016 Go to the issue

Primary Research

MiR-519d facilitates the progression and metastasis of cervical cancer through direct targeting Smad7

Retraction Note

Retraction Note to: Diagnosis, classification and grading of canine mammary tumours as a model to study human breast cancer: an Clinico-Cytohistopathological study with environmental factors influencing public health and medicine

Primary research

Transcription factor CCAAT/enhancer binding protein alpha up-regulates microRNA let-7a-1 in lung cancer cells by direct binding

Hypothesis

Comprehensive analysis of the long noncoding RNA HOXA11-AS gene interaction regulatory network in NSCLC cells

Hypothesis

Endogenous electromagnetic forces emissions during cell respiration as additional factor in cancer origin

Primary Research

SOX17 increases the cisplatin sensitivity of an endometrial cancer cell line
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

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
Image Credits