Chlorophyllin Suppresses Growth, MMP Secretion, Invasion and Cell Migration of Fibrosarcoma Cell Line HT-1080
Main Article Content
Abstract
Fibrosarcoma is an aggressive and highly malignant cancer of connective tissue with the lung being the most common site of metastasis. Surgery, chemotherapy, and radiation are the mainstay of treatments, yet the prognosis is very poor. A number of plant-based phytochemicals are increasingly being used as important treatment methods of cancers. Chlorophyll is a natural pigment that imparts the green color on plants. Chlorophyllin is a water soluble mixture of sodium-copper salts derived from chlorophyll. Chlorophyllin has been studied for its antioxidant potential. In the current study we tested the effects of chlorophyllin in fibrosarcoma HT-1080 cells on cell proliferation by MTT assay, modulation of matrix metalloproteinase (MMP) expression by zymography, cell invasive potential through Matrigel, cell migration by scratch test and morphology and apoptosis by H&E staining. Human fibrosarcoma cells HT-1080 were cultured in the media and were treated with chlorophyllin concentration at 10, 25 and 50 µM. The HT-1080 cell proliferation was significantly decreased at 50 µM dose of chlorophyllin. Expression of both, MMP-2, and MMP-9 decreased in a dose dependent manner. Both the MMPs were significantly inhibited at 25 µM and virtually undetectable at 50 µM. Cell invasion through Matrigel and cell migration was also reduced with the increasing concentrations of chlorophyllin with total inhibition of invasion at 50 µM. H&E staining at 10 µM of chlorophyllin showed a few cellular changes characteristic to apoptosis, while significant changes pertaining to apoptosis morphology were observed with increasing doses of chlorophyllin. Our results suggest that chlorophyllin may be a new chemotherapeutic strategy for fibrosarcoma patients and deserves further investigation as a potential agent in the treatment of this malignancy.
Article Details
The Medical Research Archives grants authors the right to publish and reproduce the unrevised contribution in whole or in part at any time and in any form for any scholarly non-commercial purpose with the condition that all publications of the contribution include a full citation to the journal as published by the Medical Research Archives.
References
2. https://www.cancer.org/cancer/soft-tissue-sarcoma/about/key-statistics.html
3. Ferrari A, Casanova M, Bisogno G, Cecchetto G, Meazza C, Gandola L, et al Malignant vascular tumors in children and adolescents: a report from the Italian and German Soft Tissue Sarcoma Cooperative Group. Med Pediatr Oncol 2002; 39(2): 109–114. DOI:10.1002/mpo.10078
4. Collin CF, Friedrich C, Godbold J, Hajdu S, Brennan MF. Prognostic factors for local recurrence and survival in patients with localized extremity soft-tissue sarcoma. Semin Surg Oncol 1988; 4(1): 30–37
5. Weitz J, Antonescu CR, Brennan MF. Localized extremity soft tissue sarcoma: improved knowledge with unchanged survival over time. J Clin Oncol 2003; 15(14): 2719–2725. DOI: 10.1200/JCO.2003.02.026
6. Sudakin DL. Dietary aflatoxin exposure and chemoprevention of cancer: a clinical review. J Toxicol Clin Toxicol. 2003;41(2):195-204.
7. Dashwood RH. The importance of using pure chemicals in (anti) mutagenicity studies: chlorophyllin as a case in point. Mutat Res. 1997;381(2):283-286.
8. Kumar SS, Devasagayam TP, Bhushan B, Verma NC. Scavenging of reactive oxygen species by chlorophyllin: an ESR study. Free Radic Res. 2001;35(5):563-574.],
9. Kamat JP, Boloor KK, Devasagayam TP. Chlorophyllin as an effective antioxidant against membrane damage in vitro and ex vivo. Biochim Biophys Acta. 2000;1487(2-3):113-127.
10. Tachino N, Guo D, Dashwood WM, Yamane S, Larsen R, Dashwood R. Mechanisms of the in vitro antimutagenic action of chlorophyllin against benzo[a]pyrene: studies of enzyme inhibition, molecular complex formation and degradation of the ultimate carcinogen. Mutat Res. 1994;308(2):191-203.
11. Dashwood R, Yamane S, Larsen R. Study of the forces of stabilizing complexes between chlorophylls and heterocyclic amine mutagens. Environ Mol Mutagen. 1996;27(3):211-218. DOI:10.1002/(SICI)1098-2280(1996)27:3<211::AID-EM6>3.0.CO;2-H
12. Breinholt V, Schimerlik M, Dashwood R, Bailey G. Mechanisms of chlorophyllin anticarcinogenesis against aflatoxin B1: complex formation with the carcinogen. Chem Res Toxicol. 1995;8(4):506-514.
13. Fang W, Li H, Kong L, Niu G, Gao Q, Zhou K, et al.,. Role of matrix metalloproteinases (MMPs) in tumor invasion and metastasis: serial studies on MMPs and TIMPs. Beijing Da Xue Xue Bao Yi Xue Ban. 2003;35(4):441-3. [Article in Chinese]
14. van der Horst G, Bos L, van der Pluijm G. Epithelial plasticity, cancer stem cells, and the tumor-supportive stroma in bladder carcinoma. Mol Cancer Res. Aug 2012; 10(8):995-1009. doi: 10.1158/1541-7786.MCR-12-0274
15. Giambernardi TA, Grant GM, Taylor GP, Hay RJ, Maher VM, McCormick JJ, et al,. Overview of matrix metalloproteinase expression in cultured human cells. Matrix biology. 1998;16:483–496.
16. Sternlicht MD, Werb Z. How matrix metalloproteinases regulate cell behavior. Annual review of cell and developmental biology. 2001:17463–516. DOI:10.1146/annurev.cellbio.17.1.463
17. Roomi MW, Ivanov V, Kalinovsky T, Niedzwiecki A, Rath M. In Vivo and In Vitro Antitumor Effect of Ascorbic Acid, Lysine, Proline, Arginine and Green Tea Extract on Human Fibrosarcoma Cells HT-1080, Medical Oncology 2006, 23(1): 105-112
18. Zhang YL, Guan L, Zhou PH, Mao LJ, Zhao ZM, Li SQ, Xu et al., The protective effect of chlorophyllin against oxidative damage and its mechanism. Zhonghua Nei Ke Za Zhi. 2012;51(6):466-70.[Article in Chinese]
19. Waters MD1, Stack HF, Jackson MA, Brockman HE, De Flora S. Activity profiles of antimutagens: in vitro and in vivo data. Mutat Res.1996;19;350(1):109-29.
20. Breinholt V, Schimerlik M, Dashwood R, Bailey G. Mechanisms of chlorophyllin anticarcinogenesis against aflatoxin B1: complex formation with the carcinogen. Chem Res Toxicol. 1995;8(4):506-14.
21. Simonich MT, McQuistan T, Jubert C, et al. Low-dose dietary chlorophyll inhibits multi-organ carcinogenesis in the rainbow trout. Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association. 2008;46(3):1014-1024. doi:10.1016/j.fct.2007.10.034. DOI:10.1016/j.fct.2007.10.034
22. Simonich MT, Egner PA, Roebuck BD, Orner GA, Jubert C, Pereira C, et al. Natural chlorophyll inhibits aflatoxin B1-induced multi-organ carcinogenesis in the rat. Carcinogenesis. 2007 Jun;28(6):1294-302. DOI:10.1093/carcin/bgm027
23. Vidya Priyadarsini R, Kumar N, Khan I, Thiyagarajan P, Kondaiah P, Nagini S. Gene expression signature of DMBA-induced hamster buccal pouch carcinomas: modulation by chlorophyllin and ellagic acid. PLoS One. 2012;7(4):e34628. doi: 10.1371/journal.pone.0034628.
24. Abraham SK, Sarma L, Kesavan PC. Role of chlorophyllin as an in vivo anticlastogen: protection against gamma-radiation and chemical clastogens. Mutat Res. 1994,:322(3):209-12.
25. Kumar SS, Shankar B, Sainis KB. Effect of chlorophyllin against oxidative stress in splenic lymphocytes in vitro and in vivo. Biochim Biophys Acta. 2004;3;1674(2):100-11. DOI: 10.1016/j.bbagen.2004.03.002
26. Boloor KK, Kamat JP, Devasagayam TP. Chlorophyllin as a protector of mitochondrial membranes against gamma-radiation and photosensitization.Toxicology. Nov 2000;155(1-3):63-71
27. Guo D, Schut HA, Davis CD, Snyderwine EG, Bailey GS, Dashwood RH. Protection by chlorophyllin and indole-3-carbinol against 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced DNA adducts and colonic aberrant crypts in the F344 rat. Carcinogenesis. 1995;16(12):2931-7.
28. Smith WA, Freeman JW, Gupta RC. Effect of chemopreventive agents on DNA adduction induced by the potent mammary carcinogen dibenzo[a,l]pyrene in the human breast cells MCF-7.Mutat Res. 2001;1;480-481:97-108
29. Chiu LC, Kong CK, Ooi VE. The chlorophyllin-induced cell cycle arrest and apoptosis in human breast cancer MCF-7 cells is associated with ERK deactivation and Cyclin D1 depletion. Int J Mol Med. 2005 Oct;16(4):735-40.
30. Díaz GD, Li Q, Dashwood RH. Caspase-8 and apoptosis-inducing factor mediate a cytochrome c-independent pathway of apoptosis in human colon cancer cells induced by the dietary phytochemical chlorophyllin.Cancer Res. 2003;15;63(6):1254-61