THE N-HEXANE FRACTION OF MYRMECODIA PENDANS INHIBITS CELL SURVIVAL AND PROLIFERATION IN COLON CANCER CELL LINE
Objective: Despite advanced treatment options available for colorectal cancer, many reported resistance and unresponsiveness to conventional chemotherapeutic agents. Therefore, it is urgent to discover a novel drug for colon cancer. Sarang Semut (Myrmecodia pendans), an Indonesian native plant, has been studied extensively due to its anti-cancer profiles. This study aimed to evaluate the anti-tumour activity of Sarang Semut in colon cancer cells.
Methods: We evaluated cytotoxic activity of methanol extract as well as n-hexane and ethyl acetate fraction towards colon cancer cell lines (Caco-2 and HCT-116 cells) utilizing 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The most potent fraction was evaluated further in inhibiting cell survival using MTT assay and cell proliferation using trypan blue exclusion assay as well as a clonogenic assay.
Results: Our data showed that the n-hexane fraction of Sarang Semut induces more cell death than the methanol extract and ethyl acetate fraction. Therefore, we analyzed the n-hexane fraction further and found that the inhibitory concentration 50% (IC50) of the n-hexane fraction was 24 and 30 parts per million (ppm) for Caco-2 and HCT-116 cells, respectively. Moreover, it inhibited cell growth as well as cell colony formation, in particular, shown by the plating efficiency (P<0.05) and colony area per seed (P<0.01) of the control group were different to the treatment group.
Conclusion: The n-hexane fraction of Sarang Semut demonstrates a high potential antitumor activity in colon cancer cell line.
2. Ferlay JSI, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: IARC Cancer Base No. 11:GLOBOCAN 2012 v1.0; 2012. Available from: http://globocan.iarc.fr. [Last accessed on 28 Apr 2017]
3. Andre T, Tournigand C, Achille E, Tubiana Mathieu N, Lledo G, Raoul Y, et al. [Adjuvant treatment of colon cancer MOSAIC study's main results]. Bull Cancer 2006;93 Suppl 1:S5-9.
4. Papamichael D, Audisio RA, Glimelius B, de Gramont A, Glynne-Jones R, Haller D, et al. Treatment of colorectal cancer in older patients: international society of geriatric oncology (SIOG) consensus recommendations. Ann Oncol 2015;26:463-76.
5. Hammond WA, Swaika A, Mody K. Pharmacologic resistance in colorectal cancer: a review. Ther Adv Med Oncol 2016;8:57-84.
6. Hertiani T, Sasmito E, Sumardi, Ulfah M. Preliminary study on the immunomodulatory effect of sarang-semut tubers Myrmecodia tuberosa and Myrmecodia pendens. Online J Biol Sci 2010;10:136-41.
7. Sudiono J, Oka CT, Trisfilha P. The scientific base of Myrmecodia pendans as herbal remedies. Br J Med Med Res 2015;8:230-7.
8. Soeksmanto A, Subroto MA, Wijaya H, Simanjuntak P. Anticancer activity test for extracts of sarang semut plant (Myrmecodya pendens) to HeLa and MCM-B2 cells. Pak J Biol Sci 2010;13:148-51.
9. Achmad H, Supriatno S, Marhamah M, Rasmidar R. Anti-cancer and anti-proliferation activity of ethanol fraction of ant nest plants (Myrmecodya pendans) on human tongue cancer cell SP-C1. IOSR J Dental Med Sci 2014;13:1-5.
10. Yuletnawati SE, Meiyanto E, Agustina D. High antitumor activity of ethanolic extracts of papuaâ€™s ant nest plant (Myrmecodia tuberosa) on an oral carcinoma (KB) cell line. Int J Sci Res 2016;5:1619-23.
11. Suryawati, Suardi HN, Frengki. editors. The potential antioxidant activity of ethanolic extract of aceh ant-plant (Mymercodia sp) on the free radical DPPH (1, 1-Diphenyl-2-pikrylhidrazil). Proceedings of The Annual International Conference, Syiah Kuala University-Life Sciences and Engineering Chapter; 2013.
12. Chahar MK, Sharma N, Dobhal MP, Joshi YC. Flavonoids: a versatile source of anticancer drugs. Pharmacogn Rev 2011;5:1-12.
13. Assi M, Usta J, Mounimne Y, Aboul Ela M, El Lakany A. Phytochemical study and the antiproliferative activity of Inula vulgaris species grown in Lebanon. Int J Pharm Pharm Sci 2017;9:75-83.
14. Rezano A, Kuwahara K, Yamamoto-Ibusuki M, Kitabatake M, Moolthiya P, Phimsen S, et al. Breast cancers with a high DSS1 expression that potentially maintains BRCA2 stability have poor prognosis in the relapse-free survival. BMC Cancer 2013;13:562.
15. Vallet S, Bashari MH, Fan FJ, Malvestiti S, Schneeweiss A, Wuchter P, et al. Pre-osteoblasts stimulate migration of breast cancer cells via the HGF/MET pathway. PLoS One 2016;11:e0150507.
16. Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol 2015;111:A3, B1-3.
17. Franken NA, Rodermond HM, Stap J, Haveman J, Van Bree C. Clonogenic assay of cells in vitro. Nat Protoc 2006;1:2315-9.
18. Guzman C, Bagga M, Kaur A, Westermarck J, Abankwa D. Colony area: an ImageJ plugin to automatically quantify colony formation in clonogenic assays. PLoS One 2014;9:e92444.
19. Brink M, de Goeij AF, Weijenberg MP, Roemen GM, Lentjes MH, Pachen MM, et al. K-ras oncogene mutations in sporadic colorectal cancer in the netherlands cohort study. Carcinogenesis 2003;24:703-10.
20. Shirasawa S, Furuse M, Yokoyama N, Sasazuki T. Altered growth of human colon cancer cell lines disrupted at activated Ki-ras. Science 1993;260:85-8.
21. Phipps AI, Buchanan DD, Makar KW, Win AK, Baron JA, Lindor NM, et al. KRAS-mutation status in relation to colorectal cancer survival: the joint impact of correlated tumour markers. Br J Cancer 2013;108:1757-64.
22. Engida AM, Ju YH. Recovery of antioxidants from Myrmecodia pendans and identification of its major constituent. J Anal Bioanal Tech 2013;2:3.
23. Dirgantara S, Dewi K, Raya J, Simanjuntak T. Studi botani dan fitokimia tiga spesies tanaman sarang semut asal Kabupaten Merauke, Papua. J Pharm Sci Pharm Pract 2015;2:20-3.
24. Engida AM, Kasim NS, Tsigie YA, Ismadji S, Huynh LH, Ju YH. Extraction, identification and quantitative HPLC analysis of flavonoids from sarang semut (Myrmecodia pendan). Ind Crops Prod 2013;41:392-6.
25. Supriatno M. Antitumor activity of papuaâ€™s Myrmecodia pendans in human oral tongue squamous cell carcinoma cell line through induction of cyclin-dependent kinase inhibitor p27Kip1 and suppression of cyclin E. J Cancer Res Ther 2014;2:48-53.
26. Hasanuddin, Krisnandi S, Supriadi G, Kurnia D, Adhita D. Potential of terpenoid bioactive compound isolated from papua ant nest as an alternative ovarian cancer treatment. Open J Obstetrics Gynecol 2015;5:406-11.
27. Munshi A, Hobbs M, Meyn RE. Clonogenic cell survival assay. Methods Mol Med 2005;110:21-8.