Curcumin induces re‑expression of BRCA1 and suppression of γ synuclein by modulating DNA promoter methylation in breast cancer cell lines

Al‑Yousef,Nujoud; Shinwari,Zakia; Al‑Shahrani,Bushra; Al‑Showimi,Maram; Al‑Moghrabi,Nisreen

doi:10.3892/or.2020.7473

Curcumin induces re‑expression of BRCA1 and suppression of γ synuclein by modulating DNA promoter methylation in breast cancer cell lines

Authors:
- Nujoud Al‑Yousef
- Zakia Shinwari
- Bushra Al‑Shahrani
- Maram Al‑Showimi
- Nisreen Al‑Moghrabi
View Affiliations

Affiliations: Cancer Epigenetics Section, Department of Molecular Oncology, King Faisal Specialist Hospital and Research Centre, Riyadh 11211, Kingdom of Saudi Arabia
Published online on: January 20, 2020 https://doi.org/10.3892/or.2020.7473
Pages: 827-838
Copyright: © Al‑Yousef et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Restoration of normal DNA promoter methylation and expression states of cancer‑related genes may be an option for the prevention as well as the treatment of several types of cancer. Constitutional promoter methylation of BRCA1 DNA repair associated (BRCA1) gene is linked with a high risk of developing breast and ovarian cancer. Furthermore, hypomethylation of the proto‑oncogene γ synuclein (SNCG) is associated with the metastasis of breast and ovarian cancer and reduced disease‑free survival (DFS). In the present study, we evaluated the potential of curcumin to re‑express hypermethylated BRCA1 and to suppress hypomethylated SNCG in triple‑negative breast cancer (TNBC) cell line HCC‑38, the estrogen receptor‑negative/progesterone receptor‑negative (ER‑/PR‑) cell line UACC‑3199, and the ER+/PR+ cell line T47D. The cells were treated with 5 and 10 µM curcumin for 6 days and with 5‑aza‑2'‑deoxycytidine (5'‑aza‑CdR) for 48 h. Methylation‑specific PCR and bisulfite pyrosequencing assays were used to assess DNA promoter methylation while gene expression levels were analyzed using quantitative real‑time PCR and immunoblotting. We found that curcumin treatment restored BRCA1 gene expression by reducing the DNA promoter methylation level in HCC‑38 and UACC‑3199 cells and that it suppressed the expression of SNCG by inducing DNA promoter methylation in T47D cells. Notably, 5'‑aza‑CdR restored BRCA1 gene expression only in UACC‑3199, and not in HCC‑38 cells. Curcumin‑induced hypomethylation of the BRCA1 promoter appears to be realized through the upregulation of the ten‑eleven translocation 1 (TET1) gene, whereas curcumin‑induced hypermethylation of SNCG may be realized through the upregulation of the DNA methyltransferase 3 (DNMT3) and the downregulation of TET1. Notably, miR‑29b was found to be reversely expressed compared to TET1 in curcumin‑ and 5'‑aza‑CdR‑treated cells, suggesting its involvement in the regulation of TET1. Overall, our results indicate that curcumin has an intrinsic dual function on DNA promoter methylation. We believe that curcumin may be considered a promising therapeutic option for treating TNBC patients in addition to preventing breast and ovarian cancer, particularly in cancer‑free females harboring methylated BRCA1.

View Figures

View References

1	Doll R and Peto R: The causes of cancer: Quantitative estimates of avoidable risks of cancer in the United States today. J Natl Cancer Inst. 66:1191–1308. 1981. View Article : Google Scholar : PubMed/NCBI
2	Suzuki K, Suzuki I, Leodolter A, Alonso S, Horiuchi S, Yamashita K and Perucho M: Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage. Cancer Cell. 9:199–207. 2006. View Article : Google Scholar : PubMed/NCBI
3	Goll MG and Bestor TH: Eukaryotic cytosine methyltransferases. Annu Rev Biochem. 74:481–514. 2005. View Article : Google Scholar : PubMed/NCBI
4	Ito S, Shen L, Dai Q, Wu SC, Collins LB, Swenberg JA, He C and Zhang Y: Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 5-carboxylcytosine. Science. 333:1300–1303. 2011. View Article : Google Scholar : PubMed/NCBI
5	Maiti A and Drohat AC: Thymine DNA glycosylase can rapidly excise 5-formylcytosine and 5-carboxylcytosine: Potential implications for active demethylation of CpG sites. J Biol Chem. 286:35334–35338. 2011. View Article : Google Scholar : PubMed/NCBI
6	Tahiliani M, Koh KP, Shen Y, Pastor WA, Bandukwala H, Brudno Y, Agarwal S, Iyer LM, Liu DR, Aravind L and Rao A: Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Science. 324:930–935. 2009. View Article : Google Scholar : PubMed/NCBI
7	Scourzic L, Mouly E and Bernard OA: TET proteins and the control of cytosine demethylation in cancer. Genome Med. 7:92015. View Article : Google Scholar : PubMed/NCBI
8	Good CR, Panjarian S, Kelly AD, Madzo J, Patel B, Jelinek J and Issa JJ: TET1-mediated hypomethylation activates oncogenic signaling in triple-negative breast cancer. Cancer Res. 78:4126–4137. 2018. View Article : Google Scholar : PubMed/NCBI
9	Hsu CH, Peng KL, Kang ML, Chen YR, Yang YC, Tsai CH, Chu CS, Jeng YM, Chen YT, Lin FM, et al: TET1 suppresses cancer invasion by activating the tissue inhibitors of metalloproteinases. Cell Rep. 2:568–579. 2012. View Article : Google Scholar : PubMed/NCBI
10	Yan B, Guo Q, Fu FJ, Wang Z, Yin Z, Wei YB and Yang JR: The role of miR-29b in cancer: Regulation, function, and signaling. Onco Targets Ther. 8:539–548. 2015.PubMed/NCBI
11	Zhang Z, Cao Y, Zhai Y, Ma X, An X, Zhang S and Li Z: MicroRNA-29b regulates DNA methylation by targeting Dnmt3a/3b and Tet1/2/3 in porcine early embryo development. Dev Growth Differ. 60:197–204. 2018. View Article : Google Scholar : PubMed/NCBI
12	Chou J, Lin JH, Brenot A, Kim JW, Provot S and Werb Z: GATA3 suppresses metastasis and modulates the tumour microenvironment by regulating microRNA-29b expression. Nat Cell Biol. 15:201–213. 2013. View Article : Google Scholar : PubMed/NCBI
13	Wang H, An X, Yu H, Zhang S, Tang B, Zhang X and Li Z: MiR-29b/TET1/ZEB2 signaling axis regulates metastatic properties and epithelial-mesenchymal transition in breast cancer cells. Oncotarget. 8:102119–102133. 2017.PubMed/NCBI
14	Wong EM, Southey MC, Fox SB, Brown MA, Dowty JG, Jenkins MA, Giles GG, Hopper JL and Dobrovic A: Constitutional methylation of the BRCA1 promoter is specifically associated with BRCA1 mutation-associated pathology in early-onset breast cancer. Cancer Prev Res (Phila). 4:23–33. 2011. View Article : Google Scholar : PubMed/NCBI
15	Gupta S, Jaworska-Bieniek K, Narod SA, Lubinski J, Wojdacz TK and Jakubowska A: Methylation of the BRCA1 promoter in peripheral blood DNA is associated with triple-negative and medullary breast cancer. Breast Cancer Res Treat. 148:615–622. 2014. View Article : Google Scholar : PubMed/NCBI
16	Iwamoto T, Yamamoto N, Taguchi T, Tamaki Y and Noguchi S: BRCA1 promoter methylation in peripheral blood cells is associated with increased risk of breast cancer with BRCA1 promoter methylation. Breast Cancer Res Treat. 129:69–77. 2011. View Article : Google Scholar : PubMed/NCBI
17	Al-Moghrabi N, Nofel A, Al-Yousef N, Madkhali S, Bin Amer SM, Alaiya A, Shinwari Z, Al-Tweigeri T, Karakas B, Tulbah A and Aboussekhra A: The molecular significance of methylated BRCA1 promoter in white blood cells of cancer-free females. BMC Cancer. 14:8302014. View Article : Google Scholar : PubMed/NCBI
18	Dobrovic A, Mikeska T, Alsop K, Candiloro I, George J, Mitchell G and Bowtell D: Constitutional BRCA1 methylation is a major predisposition factor for high-grade serous ovarian cancer. Cancer Res. 742014.
19	Al-Moghrabi NM: BRCA1 promoter methylation in peripheral blood cells and predisposition to breast cancer. J Taibah Univ Med Sci. 12:189–193. 2017.PubMed/NCBI
20	Wei M, Grushko TA, Dignam J, Hagos F, Nanda R, Sveen L, Xu J, Fackenthal J, Tretiakova M, Das S and Olopade OI: BRCA1 promoter methylation in sporadic breast cancer is associated with reduced BRCA1 copy number and chromosome 17 aneusomy. Cancer Res. 65:10692–10699. 2005. View Article : Google Scholar : PubMed/NCBI
21	Butcher DT, Mancini-DiNardo DN, Archer TK and Rodenhiser DI: DNA binding sites for putative methylation boundaries in the unmethylated region of the BRCA1 promoter. Int J Cancer. 111:669–678. 2004. View Article : Google Scholar : PubMed/NCBI
22	Xu J, Huo D, Chen Y, Nwachukwu C, Collins C, Rowell J, Slamon DJ and Olopade OI: CpG island methylation affects accessibility of the proximal BRCA1 promoter to transcription factors. Breast Cancer Res Treat. 120:593–601. 2010. View Article : Google Scholar : PubMed/NCBI
23	Ji H, Liu YE, Jia T, Wang M, Liu J, Xiao G, Joseph BK, Rosen C and Shi YE: Identification of a breast cancer-specific gene, BCSG1, by direct differential cDNA sequencing. Cancer Res. 57:759–764. 1997.PubMed/NCBI
24	Wu K, Quan Z, Weng Z, Li F, Zhang Y, Yao X, Chen Y, Budman D, Goldberg ID and Shi YE: Expression of neuronal protein synuclein gamma gene as a novel marker for breast cancer prognosis. Breast Cancer Res Treat. 101:259–267. 2007. View Article : Google Scholar : PubMed/NCBI
25	Lu A, Gupta A, Li C, Ahlborn TE, Ma Y, Shi EY and Liu J: Molecular mechanisms for aberrant expression of the human breast cancer specific gene 1 in breast cancer cells: Control of transcription by DNA methylation and intronic sequences. Oncogene. 20:5173–5185. 2001. View Article : Google Scholar : PubMed/NCBI
26	Lu A, Zhang F, Gupta A and Liu J: Blockade of AP1 transactivation abrogates the abnormal expression of breast cancer-specific gene 1 in breast cancer cells. J Biol Chem. 277:31364–31372. 2002. View Article : Google Scholar : PubMed/NCBI
27	Song X, Zhang M, Dai E and Luo Y: Molecular targets of curcumin in breast cancer (Review). Mol Med Rep. 19:23–29. 2019.PubMed/NCBI
28	Jiang A, Wang X, Shan X, Li Y, Wang P, Jiang P and Feng Q: Curcumin reactivates silenced tumor suppressor Gene RARβ by reducing DNA methylation. Phytother Res. 29:1237–1245. 2015. View Article : Google Scholar : PubMed/NCBI
29	Kumar U, Sharma U and Rathi G: Reversal of hypermethylation and reactivation of glutathione S-transferase pi 1 gene by curcumin in breast cancer cell line. Tumour Biol. 39:10104283176922582017. View Article : Google Scholar : PubMed/NCBI
30	Du L, Xie Z, Wu LC, Chiu M, Lin J, Chan KK, Liu S and Liu Z: Reactivation of RASSF1A in breast cancer cells by curcumin. Nutr Cancer. 64:1228–1235. 2012. View Article : Google Scholar : PubMed/NCBI
31	Zheng J, Wu C, Lin Z, Guo Y, Shi L, Dong P, Lu Z, Gao S, Liao Y, Chen B and Yu F: Curcumin up-regulates phosphatase and tensin homologue deleted on chromosome 10 through microRNA-mediated control of DNA methylation-a novel mechanism suppressing liver fibrosis. FEBS J. 281:88–103. 2014. View Article : Google Scholar : PubMed/NCBI
32	Link A, Balaguer F, Shen Y, Lozano JJ, Leung HC, Boland CR and Goel A: Curcumin modulates DNA methylation in colorectal cancer cells. PLoS One. 8:e577092013. View Article : Google Scholar : PubMed/NCBI
33	Birgisdottir V, Stefansson OA, Bodvarsdottir SK, Hilmarsdottir H, Jonasson JG and Eyfjord JE: Epigenetic silencing and deletion of the BRCA1 gene in sporadic breast cancer. Breast Cancer Res. 8:R382006. View Article : Google Scholar : PubMed/NCBI
34	Liu H, Liu W, Wu Y, Zhou Y, Xue R, Luo C, Wang L, Zhao W, Jiang JD and Liu J: Loss of epigenetic control of synuclein-gamma gene as a molecular indicator of metastasis in a wide range of human cancers. Cancer Res. 65:7635–7643. 2005. View Article : Google Scholar : PubMed/NCBI
35	Al-Moghrabi N, Al-Showimi M, Al-Yousef N, Al-Shahrani B, Karakas B, Alghofaili L, Almubarak H, Madkhali S and Al Humaidan H: Methylation of BRCA1 and MGMT genes in white blood cells are transmitted from mothers to daughters. Clin Epigenetics. 10:992018. View Article : Google Scholar : PubMed/NCBI
36	Tost J and Gut IG: DNA methylation analysis by pyrosequencing. Nat Protoc. 2:2265–2275. 2007. View Article : Google Scholar : PubMed/NCBI
37	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
38	Hendrayani SF, Al-Khalaf HH and Aboussekhra A: Curcumin triggers p16-dependent senescence in active breast cancer-associated fibroblasts and suppresses their paracrine procarcinogenic effects. Neoplasia. 15:631–640. 2013. View Article : Google Scholar : PubMed/NCBI
39	Vaughan RA, Garcia-Smith R, Dorsey J, Griffith JK, Bisoffi M and Trujillo KA: Tumor necrosis factor alpha induces Warburg-like metabolism and is reversed by anti-inflammatory curcumin in breast epithelial cells. Int J Cancer. 133:2504–2510. 2013. View Article : Google Scholar : PubMed/NCBI
40	Gupta A, Godwin AK, Vanderveer L, Lu A and Liu J: Hypomethylation of the synuclein gamma gene CpG island promotes its aberrant expression in breast carcinoma and ovarian carcinoma. Cancer Res. 63:664–673. 2003.PubMed/NCBI
41	Hu S, Xu Y, Meng L, Huang L and Sun H: Curcumin inhibits proliferation and promotes apoptosis of breast cancer cells. Exp Ther Med. 16:1266–1272. 2018.PubMed/NCBI
42	Feilotter HE, Michel C, Uy P, Bathurst L and Davey S: BRCA1 haploinsufficiency leads to altered expression of genes involved in cellular proliferation and development. PLoS One. 9:e1000682014. View Article : Google Scholar : PubMed/NCBI
43	Ma Z, Niu J, Sun E, Rong X, Zhang X and Ju Y: Gamma-synuclein binds to AKT and promotes cancer cell survival and proliferation. Tumour Biol. 37:14999–15005. 2016. View Article : Google Scholar : PubMed/NCBI
44	He JS, Xie N, Yang JB, Guan H, Chen WC, Zou C, Ouyang YW, Mao YS, Luo XY, Pan Y and Fu L: BCSG1 siRNA delivered by lentiviral vector suppressed proliferation and migration of MDA-MB-231 cells. Int J Mol Med. 41:1659–1664. 2018.PubMed/NCBI
45	Pei YF, Tao R, Li JF, Su LP, Yu BQ, Wu XY, Yan M, Gu QL, Zhu ZG and Liu BY: TET1 inhibits gastric cancer growth and metastasis by PTEN demethylation and re-expression. Oncotarget. 7:31322–31335. 2016. View Article : Google Scholar : PubMed/NCBI
46	Kai M, Niinuma T, Kitajima H, Yamamoto E, Harada T, Aoki H, Maruyama R, Toyota M, Sasaki Y, Sugai T, et al: TET1 Depletion induces aberrant CpG methylation in colorectal cancer cells. PLoS One. 11:e01682812016. View Article : Google Scholar : PubMed/NCBI
47	Chen J, Ying Y, Zhu H, Zhu T, Qu C, Jiang J and Fang B: Curcumin-induced promoter hypermethylation of the mammalian target of rapamycin gene in multiple myeloma cells. Oncol Lett. 17:1108–1114. 2019.PubMed/NCBI
48	Liu H, Zhou Y, Boggs SE, Belinsky SA and Liu J: Cigarette smoke induces demethylation of prometastatic oncogene synuclein-gamma in lung cancer cells by downregulation of DNMT3B. Oncogene. 26:5900–5910. 2007. View Article : Google Scholar : PubMed/NCBI
49	Morita S, Horii T, Kimura M, Ochiya T, Tajima S and Hatada I: miR-29 represses the activities of DNA methyltransferases and DNA demethylases. Int J Mol Sci. 14:14647–14658. 2013. View Article : Google Scholar : PubMed/NCBI