International Journal of Pharma and Bio Sciences
 
 
    ISSN 0975-6299
www.ijpbs.net


ORIGINAL RESEARCH ARTICLE
Int J Pharm Bio Sci Volume 12 Issue 1, 2021 (January-March), Pages:30-41

Evaluation of Anticancer Potential of Humic Acid and Fulvic Acid Against Breast Carcinoma

Tapasree Basu, Anil Khushalrao Shendge and Nripendranath Mandal
DOI: http://dx.doi.org/10.22376/ijpbs.2021.12.1.b30-41
Abstract:

Breast cancer is the primary cancer affecting women worldwide. Humic and fulvic acids being phenolic polymers are known for anti-inflammatory, anti-neoplastic, analgesic and anti-microbial properties. However, their role in carcinogenesis is unclear. In search of new anticancer drugs, we have extracted humic (HAF) and fulvic acids (FAF) rich fractions from dried cow dung and used for further study. HAF and FAF were characterized using FTIR analysis. We performed antioxidant and cytotoxicity assays, flow cytometry, confocal microscopy, western blotting and statistical analyses to substantiate HAF and FAF as potential antioxidant and anticancer drug candidates. It was observed that HAF and FAF were potent free radical scavengers and promising antioxidants in vitro. Both HAF and FAF triggers apoptotic cell death through selective loss of MCF-7 cell viability, morphological changes, chromosomal condensation and sub-G1 accumulation. Western blot analysis revealed the molecular markers of apoptosis associated with extrinsic and intrinsic pathways indicating ‘cytochrome C’ release from mitochondria, cleavage of pro-caspases 8, 9 and 3 followed by Bid truncation, native PARP degradation and dysregulation of Bax/Bcl-2 ratio. Further investigations showed that HAF and FAF selectively induced intracellular ROS in MCF-7 cells and suppressed the activation of ERK1/2, upregulated JNK, p38 mitogen-activated protein kinase pathways in malignant cells. Moreover p53 is upregulated in both the treatments. In this investigation, it has been reported that there is involvement of apoptotic proteins, p53 and MAP kinases which establishes the antioxidant and anti-cancer nature of HAF and FAF against MCF-7 cells. The present study will be helpful for the scientific community to develop better anticancer drug against breast carcinoma. Nevertheless, specific lead compound isolation and further in-depth study is required for development of anticancer drug.

Keywords: Humic and Fulvic acid, FT-IR, anticancer, MCF-7, MAPK, Breast Cancer
Full HTML:

REFERENCES

1.       Meiyanto E, Hermawan A, Anindyajati A. Natural products for cancer-targeted therapy: citrus flavonoids as potent chemopreventive agents. Asian Pac J Cancer Prev. 2012;13(2):427-36. doi: 10.7314/apjcp.2012.13.2.427, PMID 22524801.

2.       Globocan. Global Cancer observatory. Int Agency Res Cancer. WHO. 2018.

3.       Newman DJ, Cragg GM, Snader KM. The influence of natural products upon drug discovery. Nat Prod Rep. 2000;17(3):215-34. doi: 10.1039/a902202c, PMID 10888010.

4.       Li X, Xu H, Dai X, Zhu Z, Liu B, Lu X. Enhanced in vitro and in vivo therapeutic efficacy of codrug-loaded nanoparticles against liver cancer. Int J Nanomed. 2012;7:5183-90. doi: 10.2147/IJN.S34886, PMID 23055730.

5.       Ayurvedic formulary of India, Part I. New Delhi: Ministry of health and family welfare, govt of India 1978.

6.       Agnivesha C (with translation in Hindi, Gujarati and English). Varanasi, India 2008; Chaukhamba orientalia.

7.       Maccarthy P. The principles of humic substances. Soil Sci. 2001;166(11):738-51. doi: 10.1097/00010694-200111000-00003.

8.       Makela J, Manninen P. Humic and fulvic acids in groundwater. Earth sci. Misc Pap. 2007;23:245-52.

9.       Quanrud DM, Arnold RG, Lansey KE, Begay C, Ela W, Gandolfi AJ. Fate of effluent organic matter during soil aquifer treatment: biodegradability, chlorine reactivity and genotoxicity. J Water Health. 2003;1(1):33-44. doi: 10.2166/wh.2003.0005, PMID 15384271.

10.     Kumada K. Studies on the colour of humic acids. Soil Sci Plant Nutr. 1965;11(4):11-6. doi: 10.1080/00380768.1965.10433805.

11.     Neyroud JA, Schnitzer M. The chemistry of high molecular weight fulvic acid fractions. Can J Chem. 1974;52(24):4123-32. doi: 10.1139/v74-617.

12.     Guan J, Yang H. Medical value of humic acid. Zhong Yao Cai. 1999;22(8):430-1. PMID 12908488.

13.     Cheng ML, Ho HY, Huang YW, Lu FJ, Chiu DT. Humic acid induces oxidative DNA damage, growth retardation, and apoptosis in human primary fibroblasts. Exp Biol Med (Maywood). 2003;228(4):413-23. doi: 10.1177/153537020322800412, PMID 12671186.

14.     Cornejo A, Jiménez JM, Caballero L, Melo F, Maccioni RB. Fulvic acid inhibits aggregation and promotes disassembly of tau fibrils associated with Alzheimer’s disease. J Alzheimers Dis. 2011;27(1):143-53. doi: 10.3233/JAD-2011-110623, PMID 21785188.

15.     Van Rensburg CEJ, Malfeld SCK, Dekker J. Topical application of oxifulvic acid suppresses the cutaneous immune response in mice. Drug Dev Res. 2001;53(1):29-32. doi: 10.1002/ddr.1166.

16.     Sherry L, Millhouse E, Lappin DF, Murray C, Culshaw S, Nile CJ, Ramage G. Investigating the biological properties of carbohydrate derived fulvic acid (CHD-FA) as a potential novel therapy for the management of oral biofilm infections. BMC Oral Health. 2013;13(1):47. doi: 10.1186/1472-6831-13-47, PMID 24063298.

17.     Jayasooriya RGPT, Dilshara MG, Kang CH, Lee S, Choi YH, Jeong YK, Kim GY. Fulvic acid promotes extracellular anti-cancer mediators from RAW 264.7 cells, causing to cancer cell death in vitro. Int Immunopharmacol. 2016;36:241-8. doi: 10.1016/j.intimp.2016.04.029, PMID 27177083.

18.     Barot NS, Bagla HK. Extraction of humic acid from biological matrix − dry cow dung powder. Green Chem Lett Rev. 2009;2(4):217-21. doi: 10.1080/17518250903334290.

19.     Jayaganesh S, Senthurpan VK. Extraction and characterization of humic and fulvic acids from latosols under tea cultivation in South India. Asian J of Earth Sciences. 2010;3(3):130-5. doi: 10.3923/ajes.2010.130.135.

20.     Das A, Chaudhuri D, Mandal N, Chatterjee A. Study of antioxidant and reactive oxygen species scavenging activity of the edible tuber of ”greater yam” (Dioscorea alata L.) from north-east India. Asian J Pharm Clin Res. 2012;5:74-84.

21.     Hazra B, Biswas S, Mandal N. Antioxidant and free radical scavenging activity of Spondias pinnata. BMC Complement Altern Med. 2008;8(1):63. doi: 10.1186/1472-6882-8-63, PMID 19068130.

22.     Ghate NB, Hazra B, Sarkar R, Mandal N. In vitro anticancer activity of Spondias pinnata bark on human lung and breast carcinoma. Cytotechnology. 2014;66(2):209-18. doi: 10.1007/s10616-013-9553-7, PMID 23686547.

23.     Panja S, Ghate NB, Mandal N. A microalga, Euglena tuba induces apoptosis and suppresses metastasis in human lung and breast carcinoma cells through ROS-mediated regulation of MAPKs. Cancer Cell Int. 2016;16(1):51. doi: 10.1186/s12935-016-0330-5, PMID 27366113.

24.     Chen J, Gu B, LeBoeuf EJ, Pan H, Dai S. Spectroscopic characterization of the structural and functional properties of natural organic matter fractions. Chemosphere. 2002;48(1):59-68. doi: 10.1016/s0045-6535(02)00041-3, PMID 12137058.

25.     Bold RJ, Termuhlen PM, McConkey DJ. Apoptosis, cancer and cancer therapy. Surg Oncol. 1997;6(3):133-42. doi: 10.1016/S0960-7404(97)00015-7.

26.     Kamesaki H. Mechanisms involved in chemotherapy-induced apoptosis and their implications in cancer chemotherapy. Int J Hematol. 1998;68(1):29-43. doi: 10.1016/S0925-5710(98)00038-3, PMID 9713166.

27.     Karr MC. EP1797190A1. Lone Knight Ltd; 2007 Retrieved from https:. Method for extracting fulvic acid molecules [cited 9/11/2020]. Available from: https://patents.google.com/patent/EP1797190A1.

28.     Halliwell B. Reactive oxygen species in living systems: source, biochemistry, and role in human disease. Am J Med. 1991;91(3C):14S-22S. doi: 10.1016/0002-9343(91)90279-7, PMID 1928205.

29.     Stief TW. The physiology and pharmacology of singlet oxygen. Med Hypo. 2003;60(4):567-72. doi: 10.1016/S0306-9877(03)00026-4, PMID 12615524.

30.     Aruoma OI, Halliwell B, Hoey BM, Butler J. The antioxidant action of N-acetylcysteine: its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid. Free Radic Biol Med. 1989;6(6):593-7. doi: 10.1016/0891-5849(89)90066-X, PMID 2546864.

31.     Visioli F, Bellomo G, Galli C. Free radical-scavenging properties of olive oil polyphenols. Biochem Biophys Res Commun. 1998;247(1):60-4. doi: 10.1006/bbrc.1998.8735, PMID 9636654.

32.     Huie RE, Padmaja S. The reaction of NO with superoxide. Free Radic Res Commun. 1993;18(4):195-9. doi: 10.3109/10715769309145868, PMID 8396550.

33.     Allen RT, Hunter WJ III, Agrawal DK. Morphological and biochemical characterization and analysis of apoptosis. J Pharmacol Toxicol Methods. 1997;37(4):215-28. doi: 10.1016/S1056-8719(97)00033-6, PMID 9279777.

34.     Nicholson DW. Caspase structure, proteolytic substrates, and function during apoptotic cell death. Cell Death Differ. 1999;6(11):1028-42. doi: 10.1038/sj.cdd.4400598, PMID 10578171.

35.     Tait SWG, Green DR. Mitochondria and cell death: outer membrane permeabilization and beyond. Nat Rev Mol Cell Biol. 2010;11(9):621-32. doi: 10.1038/nrm2952, PMID 20683470.

36.     Zou H, Li Y, Liu X, Wang X. An APAF-1.cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J Biol Chem. 1999;274(17):11549-56. doi: 10.1074/jbc.274.17.11549, PMID 10206961.

37.     Jacobson MD, Raff MC. Programmed cell death and Bcl-2 protection in very low oxygen. Nature. 1995;374(6525):814-6. doi: 10.1038/374814a0, PMID 7536895.

38.     Korsmeyer SJ, Yin XM, Oltvai ZN, Veis-Novack DJ, Linette GP. Reactive oxygen species and the regulation of cell death by the BCl-2 gene family. Biochim Biophys Acta. 1995;1271(1):63-6. doi: 10.1016/0925-4439(95)00011-R, PMID 7599227.

39.     Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 1995;270(5240):1326-31. doi: 10.1126/science.270.5240.1326, PMID 7481820.

40.     Wagner EF, Nebreda AR. Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 2009;9(8):537-49. doi: 10.1038/nrc2694, PMID 19629069.

41.     Haupt S, Berger M, Goldberg Z, Haupt Y. Apoptosis - the p53 network. J Cell Sci. 2003;116(20):4077-85. doi: 10.1242/jcs.00739, PMID 12972501.

[Download PDF]
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy
Pharmaceutical Fields
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmaceutics
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Novel drug delivery system
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Nanotechnology
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmacology
Welcome to IJPBS,Pharmaceutics, Novel, drug, delivery, system, Nanotechnology, Pharmacology, Pharmacognosy Pharmacognosy
© Copyright 2009-2015 IJPBS, India. All rights reserved. Specialized online journals by ubijournal. Website by Ubitech Solutions
         Home I Contact I Terms & Conditions