Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012;7(4):e33361.
doi: 10.1371/journal.pone.0033361. Epub 2012 Apr 2.

Methylsulfonylmethane suppresses breast cancer growth by down-regulating STAT3 and STAT5b pathways

Eun Joung Lim  1 , Dae Young Hong, Jin Hee Park, Youn Hee Joung, Pramod Darvin, Sang Yoon Kim, Yoon Mi Na, Tae Sook Hwang, Sang-Kyu Ye, Eon-Soo Moon, Byung Wook Cho, Kyung Do Park, Hak Kyo Lee, Taekyu Park, Young Mok Yang
Affiliations

Methylsulfonylmethane suppresses breast cancer growth by down-regulating STAT3 and STAT5b pathways

Eun Joung Lim et al. PLoS One. 2012.

Abstract

Breast cancer is the most aggressive form of all cancers, with high incidence and mortality rates. The purpose of the present study was to investigate the molecular mechanism by which methylsulfonylmethane (MSM) inhibits breast cancer growth in mice xenografts. MSM is an organic sulfur-containing natural compound without any toxicity. In this study, we demonstrated that MSM substantially decreased the viability of human breast cancer cells in a dose-dependent manner. MSM also suppressed the phosphorylation of STAT3, STAT5b, expression of IGF-1R, HIF-1α, VEGF, BrK, and p-IGF-1R and inhibited triple-negative receptor expression in receptor-positive cell lines. Moreover, MSM decreased the DNA-binding activities of STAT5b and STAT3, to the _target gene promoters in MDA-MB 231 or co-transfected COS-7 cells. We confirmed that MSM significantly decreased the relative luciferase activities indicating crosstalk between STAT5b/IGF-1R, STAT5b/HSP90α, and STAT3/VEGF. To confirm these findings in vivo, xenografts were established in Balb/c athymic nude mice with MDA-MB 231 cells and MSM was administered for 30 days. Concurring to our in vitro analysis, these xenografts showed decreased expression of STAT3, STAT5b, IGF-1R and VEGF. Through in vitro and in vivo analysis, we confirmed that MSM can effectively regulate multiple _targets including STAT3/VEGF and STAT5b/IGF-1R. These are the major molecules involved in tumor development, progression, and metastasis. Thus, we strongly recommend the use of MSM as a trial drug for treating all types of breast cancers including triple-negative cancers.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. MSM induced cytotoxicity in human breast cancer cell lines in a dose-dependent manner.
The cytotoxicity is confirmed as apoptosis through flow cytometry. A, effect of MSM on triple-negative MDA-MB 231 cells. B, effect of MSM on SK-BR3 cells. C, flow cytometry of MDA-MB 231 cells using Annexin V-FITC, propidium iodide flow cytometry.
Figure 2
Figure 2. A, change in expression of STATs, IGF-1R, VEGF, BRCA, Brk, and hormone receptors in human breast cancer cells by MSM.
MSM substantially decreased the protein expression of human breast cancer cells in a dose-dependent manner. Whereas in normal cell line MCF-10A (Panel 1), MSM did not alter the protein expression levels. B, expression of Her-2, ER-α and PR by MSM in human breast cancer cells SKBR-3, MCF-7, and T-47D. The values are means ± S.E (n = 3) after normalization to β-actin levels (internal control). Asterisks indicate a statistically significant decrease by t-test (**p<0.01). Cell lysates were separated by 10% SDS–PAGE and transferred to a nitrocellulose membrane. The membrane was blotted with the primary antibody, then stripped and reprobed with the next antibody and so on. Data are one representative of three independent experiments.
Figure 3
Figure 3. RT-PCR analysis of breast cancer cell lines revealed MSM can effectively down regulate the expression of IGF-1R and VEGF in dose dependent manner.
A, IGF-1R and VEGF level in MDA-MB 231 cells. B, IGF-1R and VEGF level in SK-BR3 cells (table 1).
Figure 4
Figure 4. DNA binding activities of STAT5 to IGF-1R and STAT3 to VEGF sites in transfected MDA-MB 231 cells were detected by EMSA.
A, binding activity of STAT5b to the STAT5 binding site (GAS-1) of IGF-1 promoter. B, binding activity of STAT3 on the VEGF promoter site. C, nuclear protein extracts separated and blotted onto nitrocellulose membrane showing decrease in the level of p-STAT3 and p-STAT5.
Figure 5
Figure 5. Activation of the STAT3/VEGF promoter related pathway in COS-7 cells by MSM.
A, change of STAT5b/IGF-1R promoter. B, change of STAT5b/HSP90α promoter and C, change of STAT3/VEGF promoter. Data represents means of at least three separate experiments. Asterisks indicate a statistically significant decrease by ANOVA-test (***p<0.001).
Figure 6
Figure 6. Methylsulfonylmethane induced migration inhibition in metastatic breast cancer cell line MDA-MB 231.
MDA-MB 231 cells (105 cells/ml) were plated into DMEM medium. After 24 h, medium was replaced with DMEM with and without MSM. Ai, MDA-MB 231 cells without MSM. Live cell image acquired after 24 h Aii, MDA-MB 231 cells in 200 mM MSM. Live cell image acquired after 24 h. Bi, MDA-MB 231 cells without MSM. Live cell image acquired after 72 h showing the morphological changes like arborization. Bii, MDA-MB 231 cells in 200 mM MSM. Live cell image acquired after 72 h showing no morphological alteration.
Figure 7
Figure 7. Breast cancer xenografts were established in mice by subcutaneous injection of MDA-MB 231 cells (1X107 cells per mouse).
A, image of tumor-xenografted nude mice model at the end of the treatment. B, tumor size growth curves during the treatment calculating the volume size of individual tumors. C, real time PCR results of IGF-1 mRNA from breast cancer xenografts. The real time PCR was performed using TaqMan Gene Expression assays for human IGF-1 and β-actin mRNAs from ABI on ABI 7900 HT Real-time PCR system. The values are means±S.E (n = 3) after normalization to β-actin mRNA levels (internal control). Asterisks indicate a statistically significant decrease by ANOVA-test (***p<0.001). D, regulation of protein expression in xenografts by MSM. E, RT-PCR analysis of IGF-1R and VEGF mRNA levels in xenografts showed the transcriptional regulation by MSM.
Figure 8
Figure 8. Immunohistochemistry studies confirmed MSM down regulates the expression of STAT5b, IGF-1R, STAT3, and VEGF.
The xenografts were sliced to 5 µm thickness and treated with primary antibodies specific for STAT3, STAT5b, VEGF, IGF-1R, and detected using secondary antibody, Alexa Fluor 488 (rabbit) and Alexa Fluor 594 (mouse) using a magnification of 400X. A, IHC specific for STAT5b/IGF-1R. Results obtained clearly shows the decrease in STATb/IGF-1R level with no much alteration in the nucleus level. B, IHC specific for STAT3/VEGF. Results obtained clearly shows the decrease in STAT3/VEGF level with fewer alterations in the nucleus level.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Gluz O, Liedtke C, Gottschalk N, Pusztai L, Nitz U, et al. Triple-negative breast cancer–current status and future directions. Ann Oncol. 2009;20:1913–27. - PubMed
    1. Anders CK, Carey LA. Biology, metastatic patterns, and treatment of patients with triple-negative breast cancer. Clin Breast Cancer. 2009;(9Suppl 2):S73–S81. - PMC - PubMed
    1. Ismail-Khan R, Bui MM. A review of triple-negative breast cancer. Cancer Control. 2010;17(3):173–6. - PubMed
    1. Chen JQ, Russo J. ERα-negative and triple negative breast cancer: Molecular features and potential therapeutic approaches. Biochem Biophys Acta. 2009;1796:162–75. - PMC - PubMed
    1. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, et al. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 1987;235:177–82. - PubMed

Publication types

MeSH terms

  NODES
admin 1
INTERN 2
Project 1
twitter 2