IN VITRO STUDIES ON THE SYNERGISTIC EFFECT OF MORINDA CITRIFOLIA L. (NONI) AND METHOTREXATE ON CYTOTOXICITY OF HELA CELL LINES

  • MHATRE BHAKTI School of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, Sector 15, CBD Belapur, Navi Mumbai – 400 614, Maharashtra, India.
  • MARAR THANKAMANI School of Biotechnology and Bioinformatics, D.Y. Patil Deemed to be University, Sector 15, CBD Belapur, Navi Mumbai – 400 614, Maharashtra, India.

Abstract

Objective: Cancer remains to be one of the leading causes of death around the world. Modern targeted therapies have undeniably improved cancer patients’ survival, but search still continues for safer and more effective drugs. This work is an attempt to assess the effectiveness of cotreatment of aqueous fruit extract of Morinda citrifolia L. (Noni) on methotrexate (MTX)-induced cytotoxicity on HeLa cancer cell line.


Methods: HeLa cells were treated with different concentrations of MTX and Noni alone and in combination for 24 and 48 h and studied for the cytotoxic effects by various approaches.


Results: There was a dose-dependent inhibition of growth when treated with MTX in a dose range of 0.045–45.4 μg/ml and Noni for a dose range of 0.3–30 mg/ml for 48 h. The IC50 for MTX was 4.45 μg/ml (1 μM/ml), and for Noni, it was 8 mg/ml while the combination exhibited a more intensive growth inhibitory effect. Our results were confirmed with phase-contrast microscopy, whereby the number of viable cells decreased with an increase in the concentration of Noni. Cells were found to have rounded up and detached from the flask indicating apoptosis. Apoptosis in HeLa cell lines was further confirmed by DNA fragmentation and flow cytometry.


Conclusion: It can be concluded that Noni may enhance MTX cytotoxicity by inducing apoptosis and cell cycle arrest. Hence, cotreatment with Noni may reduce the dosage of MTX necessary for inhibiting proliferation of malignant cells and hence decrease the toxic effects of MTX on the system.

Keywords: Morinda citrifolia L, Methotrexate, HeLa cell lines, DNA fragmentation, Flowcytometry, apoptosis.

References

1. Ruhomally Z, Somanah J, Bahorun T, Neergheen-Bhujun VS. Morinda citrifolia L. Fruit extracts modulates H2O2-induced oxidative stress in human liposarcoma SW872 cells. J Tradit Complement Med 2016;6:299-304.
2. Gupta RK, Banerjee A, Pathak S, Sharma C, Singh N. Induction of mitochondrial-mediated apoptosis by Morinda citrifolia (Noni) in human cervical cancer cells. Asian Pac J Cancer Prev 2013;14:237-42.
3. Sawada K, Noda K, Nakajima H, Shimbara N, Furuichi Y, Sugimoto M. Differential cytotoxicity of anticancer agents in pre- and post-immortal lymphoblastoid cell lines. Biol Pharm Bull 2005;28:1202-7.
4. Wang MY, Nowicki D, Anderson G, Jensen J, West B. Liver protective effects of Morinda citrifolia (Noni). Plant Foods Hum Nutr 2008;63:59 63.
5. Kumar DJ, Santhi RJ. Antioxidant and cytotoxic effects of hexane extract of Morinda pubescens leaves in human liver cancer cell line. Asian Pac J Trop Med 2012;5:362-6.
6. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
7. Gong J, Traganos F, Darzynkiewicz Z. A selective procedure for DNA extraction from apoptotic cells applicable for gel electrophoresis and flow cytometry. Anal Biochem 1994;218:314-9.
8. Buch K, Peters T, Nawroth T, Sänger M, Schmidberger H, Langguth P. Determination of cell survival after irradiation via clonogenic assay versus multiple MTT assay – a comparative study. Radiat Oncol 2012;7:1.
9. Riccardi C, Nicoletti I. Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 2006;1:1458-61.
10. Balakrishnan SK, Bhat KI. Apoptosis and flow cytometric studies of Bauhinia variegate bark extract. Asian J Pharm Clin Res 2014;7:45-7. Available from: https://www.innovareacademics.in/journals/index.php/ajpcr/article/view/793.
11. Da’i MA, Suhendi E, Meiyanto UJ, Kawaichi M. Apoptosis induction effect of curcumin and its analogs PENTAGAMAVUNON-0 and PENTAGAMAVUNON-1 on cancer cell lines. Asian J Pharm Clin Res 2017;10:373-6.
12. Kumar AG, Nandagopal S, Illanjiam S, Suganthi K, Dhanalakshmi DP, Joshua DE. Anti-proliferative activity and immunomodulator effects of Andrographis paniculata (Burm.f.) Wall. ex Nees against cancerous cell line. Pharmacia Lett 2016;8:17-22.
13. Saraswathi U, Revatni S, Jayanthi V. In vitro evaluation of antihepatocarcinogenic activity of an herbal formulation (Chathurmuka chooranam) against HEP G 2 cells. Int J Pharm Pharm Sci 2014;6:254 6.
14. Chaudhary S, Chandrashekar KS, Pai KS, Setty MM, Devkar RA, Reddy ND, et al. Evaluation of antioxidant and anticancer activity of extract and fractions of Nardostachys jatamansi DC in breast carcinoma. BMC Complement Altern Med 2015;15:50.
15. Singh H, Banerjee S, Saumen K, Chatterjee TK. Evaluation of anticancer activity of Andaman freeze dried powdered Morinda citrifolia L. fruit against Ehrlich Ascites Carcinoma (EAC) cell induced liquid and solid tumor in Swiss albino mice. JPR BioMedRx 2013;1:567-73.
16. Banu N, Pavitra S. Anticarcinogenic effect of from Morinda citrifolia L. on HEPG2 cells. Int J Pharm Bio Sci 2015;6:145-55.
17. Mir H, Riaz M, Haseebur RN, Bilal R. Inhibition of wound closure and decreased colony formation by Clerodendrum infortunatum L. in lung cancer cell line. Int J Curr Res Biosci Plant Biol 2015;2:66-73.
18. Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C. Clonogenic assay of cells in vitro. Nat Protoc 2006;1:2315-9.
19. Latifah SY, Faujan HA, Sze LP, Raha AR, Hisyam AH, Li OC. Curcumin from turmeric (Curcuma longa) induced apoptosis in human mammary carcinoma cells (MDA-MB-23). Malays J Med Health Sci 2006;2:71-7.
20. Srinivasahan V, Durairaj B. Antioxidant and free radical scavenging effect of Morinda citrifolia fruit extract. Int J Pharm Pharm Sci 2014;6:55-9.
21. Xie L, Zhao T, Cai J, Su Y, Wang Z, Dong W. Methotrexate induces DNA damage and inhibits homologous recombination repair in choriocarcinoma cells. Onco Targets Ther 2016;9:7115-22.
22. Aziz MY, Abu N, Yeap SK, Ho WY, Omar AR, Ismail NH, et al. Combinatorial cytotoxic effects of damnacanthal and doxorubicin against human breast cancer MCF-7 cells in vitro. Molecules 2016;21. pii: E1228.
23. Kma L. Roles of plant extracts and constituents in cervical cancer therapy. Asian Pac J Cancer Prev 2013;14:3429-36.
24. Sharma K, Pachauri SD, Khandelwal K, Ahmad H, Arya A, Biala P, et al. Anticancer effects of extracts from the fruit of Morinda citrifolia (Noni) in breast cancer cell lines. Drug Res (Stuttg) 2016;66:141-7.
25. Alitheen NB, Aziz YA, Abu N, Yeap SK, Ho WY, Omar AR, et al. Combinatorial cytotoxic effects of damnacanthal and doxorubicin against human breast cancer MCF-7 Cells in vitro. Molecules 2016;21:1228-30.
Statistics
36 Views | 23 Downloads
How to Cite
MHATRE BHAKTI, and MARAR THANKAMANI. “IN VITRO STUDIES ON THE SYNERGISTIC EFFECT OF MORINDA CITRIFOLIA L. (NONI) AND METHOTREXATE ON CYTOTOXICITY OF HELA CELL LINES”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 4, Mar. 2019, pp. 173-82, https://innovareacademics.in/journals/index.php/ajpcr/article/view/31290.
Section
Original Article(s)