In vitro Cytotoxic Evaluation of Ethanol Leaf Extract of Physalis angulata Linn on Some Human Carcinoma Cell Lines

Authors

  • Onyegeme-Okerenta BM Department of BioChemistry, Faculty of Science, University of Port Harcourt, Nigeria.
  • Bradshaw TD School of Pharmacy, University of Nottingham, United Kingdom.
  • Agyare C Department of Pharmaceutics, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana.
  • Spriggs KS School of Pharmacy, University of Nottingham, United Kingdom.

Keywords:

Cytotoxicity, Human Carcinoma Cell Lines, In vitro, Physalis angulata Linn

Abstract

The search for the use of herbal remedies as an alternative medicine for the treatment and management of cancer is on-going. This study evaluates the in vitro cytotoxic potential of ethanol leaf extract of P. angulata L. on some human carcinoma cell lines. Four different human cell lines: MCF 7 (human breast), C4-2WT (prostate), HT 29 and HCT 116 (colorectal) were used for this study while cytotoxicity screening methods used are - MTT (3- (4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), Methylene blue proliferation and Trypan Blue exclusion assays. The extract significantly (p<0.05) induced cytotoxicity at 50% inhibition (GI50) as follows: MCF-7 = 3.86 ?g/ml, HT 29 = 14.99 ?g/ml, HCT 116 = 15.71 ?g/ml and C4-2WT = 20.16 ?g/ml. Methylene Blue proliferation assay response of the carcinoma cell lines to ethanol leaf extract of P. angulata L. indicated a significant decrease (p<0.05) in the total number of proliferating cells in the different concentrations of the extract used when compared to the optical densities of the controls at 24 hours, 48 hours and 72 hours. Similarly, results for Trypan Blue exclusion assay showed there was a significant decrease (p<0.05) in the total number of viable cells (VC) and a significant increase (p<0.05) in the total number of non-viable cells (NVC) over a period of 72 hours. There is a correlation between the results of the cytotoxic assays which indicated that the extract induced cytotoxicity on the human carcinoma cell lines in a time and concentration-dependent manner.

References

1. World Health Organization. Cancer Fact sheet N°297. 2018. Available from: https://www.who.int/newsroom/ fact-sheets/detail/cancer.
2. National Cancer Institute. Defining Cancer. 2018. Available from: https://www.cancer.gov/.
3. Ferlay J, Soerjomataram I, Ervik M et al. The GLOBOCAN Project 2012, Cancer Incidence and Mortality Worldwide: IARC Cancer Base No. 11. International Agency for Research on Cancer, Lyon, France. 2013.
4. Stewart BW, Wild CP. World cancer report. International Agency for Research on Cancer, Lyon. 2014.
5. Ferlay J, Shin HR, Bray F et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010; 127: 2893-917.
6. Sylla BS, Wild CP. A million Africans a year dying from cancer by 2030: What can cancer research and control offer to the continent? Int J Cancer 2011; 130(2): 245- 50.
7. Ahmad I, Aquil E, Ahmad F et al. Herbal medicines: prospects and constrains in modern phytomedicine. Turning medicinal plants into drugs, Wiley-Vch. 2006.
8. Shirwaikar A, Verma R, Lobo R et al. Phyto-therapy aspects. Nat Prod Rad 2009; 8(1): 55-63.
9. Agbor G, Talla L, Ngogang J. Toxicity of herbal preparations. Cam J Ethanobot 2005; 1: 23-28.
10. Mangan Y. Cara Bijak Menaklukkan Kanker, Cetakan I, Agromedia Pustaka, Jakarta, 2003: 28-44. 11. Pietro RC, Kashima S, Sato DN et al. In vitro antimycobacterial activities of physalis angulata L. Phytomed 2000; 7(4): 335-338.
12. Januario AH, Rodrigues FE, Pietro S et al. Antimycobacterial physalins from Physalis angulata L. (Solanaceae). Phytother Res 2002; 16(5): 445-448.
13. Magalhaes HIF, Veras ML, Torres MR et al. In-vitro and In-vivo antitumour activity of physalins Band D from physalis angulata. J Pharma Pharmacol 2006; 58(2): 235-241.
14. Wen-Tsong H, Kuan-Yuh H, Hui-Yi L et al. Food Chem Toxicol 2006; 44: 974-983.
15. Van Valkenburg JLCH, Bunyapraphatsara N. Medicinal and Poisonous Plants. Prosea Foundation, Bogor, Indonesia. Plant Resources of Southeast Asia 2002; 12(2): 782.
16. Freitas TA, Rodrigues ACC, Osuna JTA. Cultivation of Physalis angulata L. and Anadenanthera colubrina [(Vell.) Brenan] species of the Brazilian semi-arid. Rev Bras Pl Med Botucatu 2006; (8): 201-204.
17. Bahuguna A, Khan I, Bajpai VK et al. MTT assay to evaluate the cytotoxic potential of a drug. Bangladesh. J Pharmacol 2017; 12: 115-118.
18. Oliver MH, Harrison NK, Bishop JE et al. A rapid and convenient assay for counting cells cultured in microwell plates: application for assessment of growth factors. J Cell Sci 1989; 92: 513-518.
19. Onyegeme-Okerenta BM, Agyare C, Bradshaw TD et al. Cytotoxic potential of ethanol extract of Parquetina nigrescens on MCF-7, C4-2WT, HT 29 and HTC 116 cell lines. Afr J Pharm Pharmacol 2018; 212(23): 310-8.
20. Karthik R. Categories analytical techniques, analyzing cells and cell populations, methods: in cytotoxicity and cell viability with MTT assay protocol. Scientific Methods Search Engine for Biology & Biochemistry, 2016.
21. Canalle R, Burim RV, Callegari JL et al. Assessment of the cytotoxic and clastogenic activities of the sesquiterpene lactone lynchnopholide in mammalian cells in vitro and in vivo. Cancer Detec Prev 2001; 25: 93-101.
22. Aruoma OI. Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutat Res 2003; 523: 9-20.
23. Navdeep S, Anisha B, Harcharan SD et al. A pharmacological comprehensive review on ‘Rassbhary’ Physalis angulata (L.). Int J Pharm Sci 2015; 7(8): 30-34.
24. Mahavorasiriku W, Viyanant V, Chaijaroenku W et al. Cytotoxic activity of Thai medicinal plants against human cholangiocarcinoma, laryngeal and hepatocarcinoma cells in vitro. BMC Complement Altern Med 2010; 10: 55.
25. Abdel-Hameed ES, Salih A, Bazaid SA et al. Phytochemical studies and evaluation of antioxidant, anticancer and antimicrobial properties of Conocarpus erectus L. growing in Taif, Saudi Arabia. Eur J Med Plants 2012; 2: 93-112.
26. Shwu-Woan L, Min-Hsiung P, Chiu-Ming C et al. A new cytotoxic Withanolide from Physalis angulata. Chem Pharm Bull 2008; 56(2): 234-236.
27. Armandari I, Palupi KD, Farida S et al. Synergistic combination of Ciplukan (Physalis angulata) herbs ethanolic extract and doxorubicin on T47D breast cancer cells. Indones. J Cancer Chemoprevent 2010; 1(1): 26-31.
28. Sutrisna EM, Indwianiastuti, Haryadi. The ethanol extract of Physalis angulata Linn Inhibits COX-2 activity in MCF-7 Cell In Vitro. International Conference: Research and Application on Traditional Complementary and Alternative Medicine in Health Care (TCAM), Surakarta, Indonesia. 2012.
29. Kheng LO, Tengku STM, Chui HL et al. Apoptotic effects of Physalis minima L. chloroform extract in human breast carcinoma T-47D cells mediated by c-myc-, p53-, and Caspase-3-dependent pathways. Integrative Cancer Therapies 2010; 9(1): 73-83.
30. Szu-Ying W, Yann-Lii L, Ya-Ling C et al. Physalin F induces cell apoptosis in human renal carcinoma cells by targeting NF-kappa B and generating reactive oxygen species. Plos One 2012; 7(7): 1-10.

Downloads

Published

2019-04-15