Curcumin causes DNA damage and affects associated protein expression in HeLa human cervical cancer cells

Shang,Hung-Sheng; Chang,Chuan-Hsun; Chou,Yu-Ru; Yeh,Ming-Yang; Au,Man-Kuan; Lu,Hsu-Feng; Chu,Yung-Lin; Chou,Hsiao-Min; Chou,Hsiu-Chen; Shih,Yung-Luen; Chung,Jing-Gung

doi:10.3892/or.2016.5002

Curcumin causes DNA damage and affects associated protein expression in HeLa human cervical cancer cells

Authors:
- Hung-Sheng Shang
- Chuan-Hsun Chang
- Yu-Ru Chou
- Ming-Yang Yeh
- Man-Kuan Au
- Hsu-Feng Lu
- Yung-Lin Chu
- Hsiao-Min Chou
- Hsiu-Chen Chou
- Yung-Luen Shih
- Jing-Gung Chung
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Affiliations: Department of Pathology, National Defense Medical Center, Division of Clinical Pathology, Tri-Service General Hospital, Taipei, Taiwan, R.O.C., Department of Surgical Oncology, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Nutrition Therapy, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Office of Director, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Orthopedics, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C., Department of Clinical Pathology, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C., International Master's Degree Program in Food Science, International College, National Pingtung University of Science and Technology, Pingtung, Taiwan, R.O.C., Department of Pathology and Laboratory Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan, R.O.C., Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
Published online on: August 4, 2016 https://doi.org/10.3892/or.2016.5002
Pages: 2207-2215

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Abstract

Cervical cancer is one of the most common cancers in women worldwide and it is a prominent cause of cancer mortality. Curcumin is one of the major compounds from Turmeric and has been shown to induce cytotoxic cell death in human cervical cancer cells. However, there is no study to show curcumin induced DNA damage action via the effect on the DNA damage and repair protein in cervical cancer cells in detail. In this study, we investigated whether or not curcumin induced cell death via DNA damage, chromatin condensation in human cervical cancer HeLa cells by using comet assay and DAPI staining, respectively, we found that curcumin induced cell death through the induction of DNA damage, and chromatin condensation. Western blotting and confocal laser microscopy examination were used to examine the effects of curcumin on protein expression associated with DNA damage, repair and translocation of proteins. We found that curcumin at 13 µM increased the protein levels associated with DNA damage and repair, such as O6-methylguanine-DNA methyltransferase, early-onset breast cancer 1 (BRCA1), mediator of DNA damage checkpoint 1, p-p53 and p-H2A.XSer140 in HeLa cells. Results from confocal laser systems microscopy indicated that curcumin increased the translocation of p-p53 and p-H2A.XSer140 from cytosol to nuclei in HeLa cells. In conclusion, curcumin induced cell death in HeLa cells via induction of DNA damage, and chromatin condensation in vitro.

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1	Bray F, Jemal A, Grey N, Ferlay J and Forman D: Global cancer transitions according to the Human Development Index (2008–2030): A population-based study. Lancet Oncol. 13:790–801. 2012. View Article : Google Scholar : PubMed/NCBI
2	Ding Y, Yang M, She S, Min H, Xv X, Ran X, Wu Y, Wang W, Wang L, Yi L, et al: iTRAQ-based quantitative proteomic analysis of cervical cancer. Int J Oncol. 46:1748–1758. 2015.PubMed/NCBI
3	Smith HO, Tiffany MF, Qualls CR and Key CR: The rising incidence of adenocarcinoma relative to squamous cell carcinoma of the uterine cervix in the United States - a 24-year population-based study. Gynecol Oncol. 78:97–105. 2000. View Article : Google Scholar : PubMed/NCBI
4	Gascoigne KE and Taylor SS: Cancer cells display profound intra- and interline variation following prolonged exposure to antimitotic drugs. Cancer Cell. 14:111–122. 2008. View Article : Google Scholar : PubMed/NCBI
5	Rein DT and Kurbacher CM: The role of chemotherapy in invasive cancer of the cervix uteri: Current standards and future prospects. Anticancer Drugs. 12:787–795. 2001. View Article : Google Scholar : PubMed/NCBI
6	Chiappedi M and Bejor M: Herbals and natural dietary supplements in psychiatric practice. Recent Patents CNS Drug Discov. 5:164–171. 2010. View Article : Google Scholar
7	Sidhu GS, Mani H, Gaddipati JP, Singh AK, Seth P, Banaudha KK, Patnaik GK and Maheshwari RK: Curcumin enhances wound healing in streptozotocin induced diabetic rats and genetically diabetic mice. Wound Repair Regen. 7:362–374. 1999. View Article : Google Scholar : PubMed/NCBI
8	Niederau C and Göpfert E: The effect of chelidonium- and turmeric root extract on upper abdominal pain due to functional disorders of the biliary system. Results from a placebo-controlled double-blind study. Med Klin (Munich). 94:425–430. 1999.In German. View Article : Google Scholar
9	Duvoix A, Blasius R, Delhalle S, Schnekenburger M, Morceau F, Henry E, Dicato M and Diederich M: Chemopreventive and therapeutic effects of curcumin. Cancer Lett. 223:181–190. 2005. View Article : Google Scholar : PubMed/NCBI
10	Thiagarajan R and Manikandan R: Antioxidants and cataract. Free Radic Res. 47:337–345. 2013. View Article : Google Scholar : PubMed/NCBI
11	Basnet P and Skalko-Basnet N: Curcumin: An anti-inflammatory molecule from a curry spice on the path to cancer treatment. Molecules. 16:4567–4598. 2011. View Article : Google Scholar : PubMed/NCBI
12	Kunnumakkara AB, Anand P and Aggarwal BB: Curcumin inhibits proliferation, invasion, angiogenesis and metastasis of different cancers through interaction with multiple cell signaling proteins. Cancer Lett. 269:199–225. 2008. View Article : Google Scholar : PubMed/NCBI
13	Shehzad A, Wahid F and Lee YS: Curcumin in cancer chemoprevention: Molecular targets, pharmacokinetics, bioavailability, and clinical trials. Arch Pharm (Weinheim). 343:489–499. 2010. View Article : Google Scholar
14	Bava SV, Puliappadamba VT, Deepti A, Nair A, Karunagaran D and Anto RJ: Sensitization of taxol-induced apoptosis by curcumin involves down-regulation of nuclear factor-kappaB and the serine/threonine kinase Akt and is independent of tubulin polymerization. J Biol Chem. 280:6301–6308. 2005. View Article : Google Scholar
15	Goel A, Kunnumakkara AB and Aggarwal BB: Curcumin as 'Curecumin': From kitchen to clinic. Biochem Pharmacol. 75:787–809. 2008. View Article : Google Scholar
16	Strimpakos AS and Sharma RA: Curcumin: Preventive and therapeutic properties in laboratory studies and clinical trials. Antioxid Redox Signal. 10:511–545. 2008. View Article : Google Scholar : PubMed/NCBI
17	Lu JJ, Cai YJ and Ding J: Curcumin induces DNA damage and caffeine-insensitive cell cycle arrest in colorectal carcinoma HCT116 cells. Mol Cell Biochem. 354:247–252. 2011. View Article : Google Scholar : PubMed/NCBI
18	Park C, Kim GY, Kim GD, Choi BT, Park YM and Choi YH: Induction of G2/M arrest and inhibition of cyclooxygenase-2 activity by curcumin in human bladder cancer T24 cells. Oncol Rep. 15:1225–1231. 2006.PubMed/NCBI
19	Tomita M, Kawakami H, Uchihara JN, Okudaira T, Masuda M, Takasu N, Matsuda T, Ohta T, Tanaka Y, Ohshiro K, et al: Curcumin (diferuloylmethane) inhibits constitutive active NF-kappaB, leading to suppression of cell growth of human T-cell leukemia virus type I-infected T-cell lines and primary adult T-cell leukemia cells. Int J Cancer. 118:765–772. 2006. View Article : Google Scholar
20	Giri AK, Das SK, Talukder G and Sharma A: Sister chromatid exchange and chromosome aberrations induced by curcumin and tartrazine on mammalian cells in vivo. Cytobios. 62:111–117. 1990.PubMed/NCBI
21	Blasiak J, Trzeciak A and Kowalik J: Curcumin damages DNA in human gastric mucosa cells and lymphocytes. J Environ Pathol Toxicol Oncol. 18:271–276. 1999.
22	Cao J, Jia L, Zhou HM, Liu Y and Zhong LF: Mitochondrial and nuclear DNA damage induced by curcumin in human hepatoma G2 cells. Toxicol Sci. 91:476–483. 2006. View Article : Google Scholar : PubMed/NCBI
23	Kim B, Kim HS, Jung EJ, Lee JY, K Tsang B, Lim JM and Song YS: Curcumin induces ER stress-mediated apoptosis through selective generation of reactive oxygen species in cervical cancer cells. Mol Carcinog. 55:918–928. 2015. View Article : Google Scholar : PubMed/NCBI
24	Lewinska A, Adamczyk J, Pajak J, Stoklosa S, Kubis B, Pastuszek P, Slota E and Wnuk M: Curcumin-mediated decrease in the expression of nucleolar organizer regions in cervical cancer (HeLa) cells. Mutat Res Genet Toxicol Environ Mutagen. 771:43–52. 2014. View Article : Google Scholar : PubMed/NCBI
25	Singh M and Singh N: Molecular mechanism of curcumin induced cytotoxicity in human cervical carcinoma cells. Mol Cell Biochem. 325:107–119. 2009. View Article : Google Scholar : PubMed/NCBI
26	Sreekanth CN, Bava SV, Sreekumar E and Anto RJ: Molecular evidences for the chemosensitizing efficacy of liposomal curcumin in paclitaxel chemotherapy in mouse models of cervical cancer. Oncogene. 30:3139–3152. 2011. View Article : Google Scholar : PubMed/NCBI
27	Huang AC, Lin TP, Weng YS, Ho YT, Lin HJ, Huang LJ, Kuo SC and Chung JG: Ethyl 2-[N-m-chlorobenzyl-(2′-methyl)] anilino-4-oxo-4,5-dihydrofuran-3-carboxylate (JOT01006) induces apoptosis in human cervical cancer HeLa cells. Anticancer Res. 27:2505–2514. 2007.PubMed/NCBI
28	Leung YM, Wong KL, Chen SW, Lu DY, Kuo CS, Chen YR, Chen YW and Cheng TH: Down-regulation of voltage-gated Ca²⁺ channels in Ca²⁺ store-depleted rat insulinoma RINm5F cells. Biomedicine. 3:130–139. 2013. View Article : Google Scholar
29	Lin MC, Tsai SY, Wang FY, Liu FH, Syu JN and Tang FY: Leptin induces cell invasion and the upregulation of matrilysin in human colon cancer cells. Biomedicine. 3:174–180. 2013. View Article : Google Scholar
30	Wu LY, Lu HF, Chou YC, Shih YL, Bau DT, Chen JC, Hsu SC and Chung JG: Kaempferol induces DNA damage and inhibits DNA repair associated protein expressions in human promyelocytic leukemia HL-60 cells. Am J Chin Med. 43:365–382. 2015. View Article : Google Scholar : PubMed/NCBI
31	Błasiak J, Trzeciak A, Małecka-Panas E, Drzewoski J, Iwanienko T, Szumiel I and Wojewódzka M: DNA damage and repair in human lymphocytes and gastric mucosa cells exposed to chromium and curcumin. Teratog Carcinog Mutagen. 19:19–31. 1999. View Article : Google Scholar
32	Ashby J, Tinwell H, Lefevre PA and Browne MA: The single cell gel electrophoresis assay for induced DNA damage (comet assay): Measurement of tail length and moment. Mutagenesis. 10:85–90. 1995. View Article : Google Scholar : PubMed/NCBI
33	Pool-Zobel BL, Lotzmann N, Knoll M, Kuchenmeister F, Lambertz R, Leucht U, Schröder HG and Schmezer P: Detection of genotoxic effects in human gastric and nasal mucosa cells isolated from biopsy samples. Environ Mol Mutagen. 24:23–45. 1994. View Article : Google Scholar : PubMed/NCBI
34	Marchissio MJ, Francés DE, Carnovale CE and Marinelli RA: Evidence for necrosis, but not apoptosis, in human hepatoma cells with knockdown of mitochondrial aquaporin-8. Apoptosis. 19:851–859. 2014. View Article : Google Scholar : PubMed/NCBI
35	Zheng B, Wu L, Ma L, Liu S, Li L, Xie W and Li X: Telekin induces apoptosis associated with the mitochondria-mediated pathway in human hepatocellular carcinoma cells. Biol Pharm Bull. 36:1118–1125. 2013. View Article : Google Scholar : PubMed/NCBI
36	Olive PL, Banáth JP and Durand RE: Detection of etoposide resistance by measuring DNA damage in individual Chinese hamster cells. J Natl Cancer Inst. 82:779–783. 1990. View Article : Google Scholar : PubMed/NCBI
37	Tice RR, Andrews PW and Singh NP: The single cell gel assay: A sensitive technique for evaluating intercellular differences in DNA damage and repair. Basic Life Sci. 53:291–301. 1990.PubMed/NCBI
38	Malhotra P, Adhikari M, Singh SK and Kumar R: N-acetyl tryptophan glucopyranoside (NATG) provides radioprotection to murine macrophage J774A.1 cells. Free Radic Res. 49:1488–1498. 2015. View Article : Google Scholar : PubMed/NCBI
39	Banzai C, Nishino K, Quan J, Yoshihara K, Sekine M, Yahata T and Tanaka K; Gynecological Cancer Registry of Niigata: Promoter methylation of DAPK1, FHIT, MGMT, and CDKN2A genes in cervical carcinoma. Int J Clin Oncol. 19:127–132. 2014. View Article : Google Scholar
40	Huang J, Ye F, Chen H, Lu W and Xie X: Amino acid substitution polymorphisms of the DNA repair gene MGMT and the susceptibility to cervical carcinoma. Carcinogenesis. 28:1314–1322. 2007. View Article : Google Scholar : PubMed/NCBI
41	Verbeek B, Southgate TD, Gilham DE and Margison GP: O⁶-Methylguanine-DNA methyltransferase inactivation and chemotherapy. Br Med Bull. 85:17–33. 2008. View Article : Google Scholar
42	Christmann M, Verbeek B, Roos WP and Kaina B: O(6)-Methylguanine-DNA methyltransferase (MGMT) in normal tissues and tumors: Enzyme activity, promoter methylation and immunohistochemistry. Biochim Biophys Acta. 1816:179–190. 2011.PubMed/NCBI
43	Dueñas-González A, Lizano M, Candelaria M, Cetina L, Arce C and Cervera E: Epigenetics of cervical cancer. An overview and therapeutic perspectives. Mol Cancer. 4:382005. View Article : Google Scholar : PubMed/NCBI
44	Abramovitch S, Glaser T, Ouchi T and Werner H: BRCA1-Sp1 interactions in transcriptional regulation of the IGF-IR gene. FEBS Lett. 541:149–154. 2003. View Article : Google Scholar : PubMed/NCBI
45	Maor SB, Abramovitch S, Erdos MR, Brody LC and Werner H: BRCA1 suppresses insulin-like growth factor-I receptor promoter activity: Potential interaction between BRCA1 and Sp1. Mol Genet Metab. 69:130–136. 2000. View Article : Google Scholar : PubMed/NCBI
46	Taron M, Rosell R, Felip E, Mendez P, Souglakos J, Ronco MS, Queralt C, Majo J, Sanchez JM, Sanchez JJ, et al: BRCA1 mRNA expression levels as an indicator of chemoresistance in lung cancer. Hum Mol Genet. 13:2443–2449. 2004. View Article : Google Scholar : PubMed/NCBI
47	Lou Z, Minter-Dykhouse K, Franco S, Gostissa M, Rivera MA, Celeste A, Manis JP, van Deursen J, Nussenzweig A, Paull TT, et al: MDC1 maintains genomic stability by participating in the amplification of ATM-dependent DNA damage signals. Mol Cell. 21:187–200. 2006. View Article : Google Scholar : PubMed/NCBI
48	Amichay K, Kidron D, Attias-Geva Z, Schayek H, Sarfstein R, Fishman A, Werner H and Bruchim I: BRCA1 is expressed in uterine serous carcinoma (USC) and controls insulin-like growth factor I receptor (IGF-IR) gene expression in USC cell lines. Int J Gynecol Cancer. 22:748–754. 2012. View Article : Google Scholar : PubMed/NCBI
49	Prives C and Hall PA: The p53 pathway. J Pathol. 187:112–126. 1999. View Article : Google Scholar : PubMed/NCBI
50	Gorgoulis VG, Zacharatos P, Mariatos G, Liloglou T, Kokotas S, Kastrinakis N, Kotsinas A, Athanasiou A, Foukas P, Zoumpourlis V, et al: Deregulated expression of c-mos in non-small cell lung carcinomas: Relationship with p53 status, genomic instability, and tumor kinetics. Cancer Res. 61:538–549. 2001.PubMed/NCBI
51	Bonner WM, Redon CE, Dickey JS, Nakamura AJ, Sedelnikova OA, Solier S and Pommier Y: GammaH2AX and cancer. Nat Rev Cancer. 8:957–967. 2008. View Article : Google Scholar : PubMed/NCBI
52	Celeste A, Petersen S, Romanienko PJ, Fernandez-Capetillo O, Chen HT, Sedelnikova OA, Reina-San-Martin B, Coppola V, Meffre E, Difilippantonio MJ, et al: Genomic instability in mice lacking histone H2AX. Science. 296:922–927. 2002. View Article : Google Scholar : PubMed/NCBI