Contribution of the ROS-p53 feedback loop in thuja-induced apoptosis of mammary epithelial carcinoma cells

Saha,Shilpi; Bhattacharjee,Pushpak; Mukherjee,Shravanti; Mazumdar,Minakshi; Chakraborty,Samik; Khurana,Anil; Nayak,Debadatta; Manchanda,Rajkumar; Chakrabarty,Rathin; Das,Tanya; Sa,Gaurisankar

doi:10.3892/or.2014.2993

Contribution of the ROS-p53 feedback loop in thuja-induced apoptosis of mammary epithelial carcinoma cells

Authors:
- Shilpi Saha
- Pushpak Bhattacharjee
- Shravanti Mukherjee
- Minakshi Mazumdar
- Samik Chakraborty
- Anil Khurana
- Debadatta Nayak
- Rajkumar Manchanda
- Rathin Chakrabarty
- Tanya Das
- Gaurisankar Sa
View Affiliations

Affiliations: Division of Molecular Medicine, Bose Institute, P1/12, CIT Scheme VIIM, Kolkata 700054, India, Central Council for Research in Homeopathy, 61-65 Institutional Area, Janakpuri, New Delhi 110058, India, Bholanath Chakrabarty Trust, 5 Subol Koley Lane, Howrah 711101, India
Published online on: January 24, 2014 https://doi.org/10.3892/or.2014.2993
Pages: 1589-1598

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

The adverse side-effects associated with chemotherapy during cancer treatment have shifted considerable focus towards therapies that are targeted but devoid of toxic side-effects. In the present study, the antitumorigenic activity of thuja, the bioactive derivative of the medicinal plant Thuja occidentalis, was evaluated, and the molecular mechanisms underlying thuja-induced apoptosis of functional p53-expressing mammary epithelial carcinoma cells were elucidated. Our results showed that thuja successfully induced apoptosis in functional p53-expressing mammary epithelial carcinoma cells. Abrogation of intracellular reactive oxygen species (ROS), prevention of p53-activation, knockdown of p53 or inhibition of its functional activity significantly abridged ROS generation. Notably, under these conditions, thuja-induced breast cancer cell apoptosis was reduced, thereby validating the existence of an ROS-p53 feedback loop. Elucidating this feedback loop revealed bi-phasic ROS generation as a key mediator of thuja-induced apoptosis. the first phase of ROS was instrumental in ensuring activation of p53 via p38MAPK and its nuclear translocation for transactivation of Bax, which induced a second phase of mitochondrial ROS to construct the ROS-p53 feedback loop. Such molecular crosstalk induced mitochondrial changes i) to maintain and amplify the thuja signal in a positive self-regulatory feedback manner; and ii) to promote the mitochondrial death cascade through cytochrome c release and caspase-driven apoptosis. These results open the horizon for developing a targeted therapy by modulating the redox status of functional p53-expressing mammary epithelial carcinoma cells by thuja.

View Figures

View References

1	Hanf V and Gonder U: Nutrition and primary prevention of breast cancer: foods, nutrients and breast cancer risk. Eur J Obstet Gynecol Reprod Biol. 123:139–149. 2005. View Article : Google Scholar : PubMed/NCBI
2	Cassileth BR and Vickers AJ: Complementary and alternative therapies. Urol Clin North Am. 30:369–376. 2003. View Article : Google Scholar
3	Flinn JE: Bromium in acute lymphatic leukemia. J Am Inst Homeopath. 58:213–214. 1965.PubMed/NCBI
4	Gruchmann W: Arsenic: destroyer and healer; a contribution to the management of carcinoma. Hippokrates. 27:444–445. 1956.(In German).
5	Pathak S, Multani AS and Banerji P and Banerji P: Ruta 6 selectively induces cell death in brain cancer cells but proliferation in normal peripheral blood lymphocytes: a novel treatment for human brain cancer. Int J Oncol. 23:975–982. 2003.PubMed/NCBI
6	Pathak S, Kumar Das J, Jyoti Biswas S and Khuda-Bukhsh AR: Protective potentials of a potentized homeopathic drug, Lycopodium-30, in ameliorating azo dye induced hepatocarcinogenesis in mice. Mol Cell Biochem. 285:121–131. 2006. View Article : Google Scholar : PubMed/NCBI
7	Frenkel M, Mishra BM, Sen S, Yang P, Pawlus A, Vence L, Leblanc A, Cohen L and Banerji P and Banerji P: Cytotoxic effects of ultra-diluted remedies on breast cancer cells. Int J Oncol. 36:395–403. 2010.PubMed/NCBI
8	Ojeswi BK, Khoobchandani M, Hazra DK and Srivastava MM: Protective effect of Thuja occidentalis against DMBA-induced breast cancer with reference to oxidative stress. Hum Exp Toxicol. 29:369–375. 2010.
9	Biswas R, Mandal SK, Dutta S, Bhattacharyya SS, Boujedaini N and Khuda-Bukhsh AR: Thujone-rich fraction of Thuja occidentalis demonstrates major anti-cancer potentials: evidences from in vitro studies on A375 cells. Evid Based Complement Alternat Med. 2011:5681482011.PubMed/NCBI
10	Naser B, Bodinet C, Tegtmeier M and Lindequist U: Thuja occidentalis (Arbor vitae): a review of its pharmaceutical, pharmacological and clinical properties. Evid Based Complement Alternat Med. 2:69–78. 2005. View Article : Google Scholar
11	Sunila ES and Kuttan G: Protective effect of Thuja occidentalis against radiation-induced toxicity in mice. Integr Cancer Ther. 4:322–328. 2005.
12	Sunila ES and Kuttan G: A preliminary study on antimetastatic activity of Thuja occidentalis L. in mice model. Immunopharmacol Immunotoxicol. 28:269–280. 2006. View Article : Google Scholar : PubMed/NCBI
13	Dubey SK and Batra A: Hepatoprotective activity from ethanol fraction of Thuja occidentalis Linn. Asian J Res Chem. 1:32–35. 2008.
14	Dubey SK and Batra A: Antioxidant activity of Thuja occidentalis Linn. Asian J Pharm Clin Res. 2:73–76. 2009.
15	Efeyan A and Serrano M: p53: guardian of the genome and policeman of the oncogenes. Cell Cycle. 6:1006–1010. 2007. View Article : Google Scholar : PubMed/NCBI
16	Liu B, Chen Y and St Clair DK: ROS and p53: a versatile partnership. Free Radic Biol Med. 44:1529–1535. 2008. View Article : Google Scholar : PubMed/NCBI
17	Pervaiz S and Clement MV: Tumor intracellular redox status and drug resistance - serendipity or a causal relationship? Curr Pharm Des. 10:1969–1977. 2004. View Article : Google Scholar : PubMed/NCBI
18	Mandal D, Lahiry L, Bhattacharyya A, Chattopadhyay S, Siddiqi M, Sa G and Das T: Black tea protects thymocytes in tumor-bearing animals by differential regulation of intracellular ROS in tumor cells and thymocytes. J Environ Pathol Toxicol Oncol. 24:91–104. 2005. View Article : Google Scholar : PubMed/NCBI
19	Adhikary A, Mohanty S, Lahiry L, Hossain DM, Chakraborty S and Das T: Theaflavins retard human breast cancer cell migration by inhibiting NF-κB via p53-ROS cross-talk. FEBS Lett. 584:7–14. 2010.PubMed/NCBI
20	Mazumdar M, Adhikary A, Chakraborty S, Mukherjee S, Manna A, Saha S, Mohanty S, Dutta A, Bhattacharjee P, Ray P, Chattopadhyay S, Banerjee S, Chakraborty J, Ray AK, Sa G and Das T: Targeting RET to induce medullary thyroid cancer cell apoptosis: an antagonistic interplay between PI3K/Akt and p38MAPK/caspase-8 pathways. Apoptosis. 18:589–604. 2013. View Article : Google Scholar : PubMed/NCBI
21	Lahiry L, Saha B, Chakraborty J, Bhattacharyya S, Chattopadhyay S, Banerjee S, Choudhuri T, Mandal D, Bhattacharyya A, Sa G and Das T: Contribution of p53-mediated Bax transactivation in theaflavin-induced mammary epithelial carcinoma cell apoptosis. Apoptosis. 13:771–781. 2008. View Article : Google Scholar : PubMed/NCBI
22	Saito S, Yamaguchi H, Higashimoto Y, Chao C, Xu Y, Fornace AJ Jr, Appella E and Anderson CW: Phosphorylation site interdependence of human p53 post-translational modifications in response to stress. J Biol Chem. 278:37536–37544. 2003. View Article : Google Scholar : PubMed/NCBI
23	Freund A, Patil CK and Campisi J: p38MAPK is a novel DNA damage response-independent regulator of the senescence-associated secretory phenotype. EMBO J. 30:1536–1548. 2011. View Article : Google Scholar : PubMed/NCBI
24	Murphy PJ, Galigniana MD, Morishima Y, Harrell JM, Kwok RP, Ljungman M and Pratt WB: Pifithrin-α inhibits p53 signaling after interaction of the tumor suppressor protein with hsp90 and its nuclear translocation. J Biol Chem. 279:30195–30201. 2004.
25	Chang LC, Song LL, Park EJ, Luyengi L, Lee KJ, Farnsworth NR, Pezzuto JM and Kinghorn AD: Bioactive constituents of Thuja occidentalis. J Nat Prod. 63:1235–1238. 2000. View Article : Google Scholar
26	British Herbal Pharmacopoeia. Thuja, British HerbalMedicine Association; West Yorks, UK: 1983
27	Thangapazham RL, Gaddipati JP, Rajeshkumar NV, Sharma A, Singh AK, Ives JA, Maheshwari RK and Jonas WB: Homeopathic medicines do not alter growth and gene expression in prostate and breast cancer cells in vitro. Integr Cancer Ther. 5:356–361. 2006. View Article : Google Scholar : PubMed/NCBI
28	MacLaughlin BW, Gutsmuths B, Pretner E, Jonas WB, Ives J, Kulawardane DV and Amri H: Effects of homeopathic preparations on human prostate cancer growth in cellular and animal models. Integr Cancer Ther. 5:362–372. 2006. View Article : Google Scholar : PubMed/NCBI
29	Preethi K, Ellanghiyil S, Kuttan G and Kuttan R: Induction of apoptosis of tumor cells by some potentiated homeopathic drugs: implications on mechanism of action. Integr Cancer Ther. 11:172–182. 2012. View Article : Google Scholar : PubMed/NCBI
30	Bishayee K, Paul A, Ghosh S, Sikdar S, Mukherjee A, Biswas R, Boujedaini N and Khuda-Bukhsh AR: Condurango-glycoside-A fraction of Gonolobus condurango induces DNA damage associated senescence and apoptosis via ROS-dependent p53 signalling pathway in HeLa cells. Mol Cell Biochem. 82:173–183. 2013.
31	Huschtscha L, Bartier W, Malmstrom A and Tattersall M: Cell-death by apoptosis following anticancer drug-treatment in vitro. Int J Oncol. 6:585–593. 1995.PubMed/NCBI
32	Gu B and Zhu WG: Surf the post-translational modification network of p53 regulation. Int J Biol Sci. 8:672–684. 2012. View Article : Google Scholar : PubMed/NCBI
33	Bulavin DV, Saito S, Hollander MC, Sakaguchi K, Anderson CW, Appella E and Fornace AJ Jr: Phosphorylation of human p53 by p38 kinase coordinates N-terminal phosphorylation and apoptosis in response to UV radiation. EMBO J. 18:6845–6854. 1999. View Article : Google Scholar : PubMed/NCBI
34	Sanchez-Prieto R, Rojas JM, Taya Y and Gutkind JS: A role for the p38 mitogen-activated protein kinase pathway in the transcriptional activation of p53 on genotoxic stress by chemotherapeutic agents. Cancer Res. 60:2464–2472. 2000.PubMed/NCBI
35	Oda K, Arakawa H, Tanaka T, Matsuda K, Tanikawa C, Mori T, Nishimori H, Tamai K, Tokino T, Nakamura Y and Taya Y: p53AIP1, a potential mediator of p53-dependent apoptosis, and its regulation by Ser-46-phosphorylated p53. Cell. 102:849–862. 2000. View Article : Google Scholar : PubMed/NCBI
36	Kim H, You S, Farris J, Foster LK and Foster DN: Post-transcriptional inactivation of p53 in immortalized murine embryo fibroblast cells. Oncogene. 20:3306–3310. 2001. View Article : Google Scholar : PubMed/NCBI