SELECTIVITY OF PURIFIED EXTRACT FROM THE LEAVES OF Tithonia diversifolia (Hemsley) A.Gray AGAINST HELA CELLS

https://doi.org/10.22146/tradmedj.7757

M. Sri Hartati Wahyuningsih(1*), Rul Afiyah Syarif(2), Sri Suharmi(3), Tri Murini(4), Firandi Saputra(5), Adiguno Suryo W(6)

(1) Faculty of Medicine, Universitas Gadjah Mada
(2) Faculty of Medicine, Universitas Gadjah Mada
(3) Faculty of Medicine, Universitas Gadjah Mada
(4) Faculty of Medicine, Universitas Gadjah Mada
(5) Faculty of Medicine, Universitas Gadjah Mada
(6) Faculty of Medicine, Universitas Gadjah Mada
(*) Corresponding Author

Abstract


Tithonia diversifolia (Hemsley) A.Gray is a kind of plant and traditionally used to cure various diseases. The previous research found that T.diversifolia had antiproliferation effect on colon cancer cell (Col2). Sample preparation using biossay guided extraction and partition method, could simplify the active compounds of the extract. Research on the selectivity of purified extract from the leaves of Tithonia diversifolia (Hemsley) A.Gray) against HeLa cells hasn’t been known yet. The aim of this research was to find out the selectivity of purified extract of T.diversifolia’s leaves on HeLa cell compared to vero cell and to find out the Fifty Percent Inhibition Concentration (IC50), and its selectivity index. Extraction of T. diversifolia‘s leaves were done by maceration method using chloroform and methanol. Both of extract were tested by MTT cytotoxic assay on HeLa cells in vitro with serial doses (0,24 - 500μg/mL). Each group was replicated 3 times. The absorbance was read using ELISA at λ 540nm. IC50 was analyzed by probit regression on SPSS 15 for Windows. Partition (purified extract) of the active extract was done using Petroleum eter (PE), and tested by MTT cytotoxic assay on HeLa cells in vitro.  The IC50 methanol extract to HeLa cell in vitro is 1006,99μg/mL, IC50 chloroform extract is 16,61μg/mL.  The IC50 of soluble PE is 325,33μg/mL.  and IC50 of insoluble PE is 3,078μg/mL. The IC50 value of insoluble PE to Vero cell is 80,30 μg/mL. Selectivity index of purified extract (PE insoluble extract) is 26.09. 


Keywords


Extraction, purification, T. diversifolia, HeLa cell, IC50



References

Alliance for Cervical Cancer Prevention (ACP), 2004. Preventing Cervical Cancer World Wide. Population Reference Bureque, Washington

Baruah NC, Sharma RP, Madhusudanan KP, Thyagarajan G, 1979, Sesquiterpene

Lactones of Tithonia diversifolia. Stereochemistry of the Tagitinins and Related Compounds. J. Org. Chem. 44(11):1831

Benzivin C, Devehat, Tomasi S, Boustie J, 2003, Cytotoxic Activities of some Lischen Extracts on Murine and Humasn Cancer Cell Lines. Phytomedicine, 10:499-503

Boyd, M.R., 1997. The NCI In Vitro Anticancer Drug Discovery Screen. Concept, Implementation, and Operation, 1985-1995. Dalam: Teicher, B.(ed.): Drug Development: Preclinical Screening, Clinical Trials and Approval. Humana Press,Totowa. p. 23-42

DeVita,V.T., Hellman,S., dan Rosenberg,S.A. (Eds.), 2001. Cancer: Principles and Practices of Oncology. 6th ed. Lippincot William ans Wilkins, Philadelphia

Freshney,R.I., 2000. Culture of Animal Cells Manual of Basic Technique. A John Wiley and Sons Ltd, New York

Ganiswara, S., dan Nafrialdi. 1995. Antikanker dan Immunosupresan, dalam Ganiswara, S., (Ed), Farmakologi dan Terapi, Bagian Farmakologi Fakultas Kedokteran, Universitas Indonesia

García A, dan Delgado G, 2006, Constituents from Tithonia diversifolia. Stereochemical Revision of 2α-Hydroxytirotundin. J. Mex. Chem. Soc. 50(4):180-3

Goffin,E., Ziemons,E., De Mol,P., de Madureira Mdo,C., Martins,A.P., da Cunha,A.P., Philippe,G., Tiys,M., Angenot,L., dan Frederich,M., 2002. In vitro antiplasmodial activity of Tithonia diversifolia and identification of its main active constituent: Tagitin C. Planta Medica 68(6):543-545

Gu,J.Q., Gills,J.J., Park,E.J., Mata-Greenwood,E., Hawthorne,M.E., Axelrod,F., Chavez,P.I., Fong,H.H., Mehta,R.G., Pezzuto,J.M., dan Kinghorn,A.D., 2002. Sesquiterpenoids from Tithonia diversifolia with potential care chemo preventive activity. J Nat Prod 65(4),532-536

Hutapea JR, 1994, Inventaris Tanaman Obat Indonesia. Badan Peneliti dan Pengembangan Kesehatan R.I., Jakarta

Madureira, M. C., Martins, A. P., Gomes, M., Paiva, J., Cunha, A. P., dan Rosario, V., 2002. Antimalarial activity of medicinal plants used in traditional medicine in S. Tome and Principe islands. J. Ethnopharmacol 81:23 – 9

Miura, T., Furuta, K., Yasuda, A., Iwamoto, N., Kato, M., Ishihara, E., Ishida, T., dan Tanigawa, K.,2002. Antidiabetic effect of Nitobegiku in KK-Ay diabetic mice. The American Journal of Chinese Medicine 30 (1):81–6

Moronkola,D.O., Ogunwade,I.A., Walker,T.M., Setzer,W.N., dan Oyewole,I.O., 2006. Identification of the main volatile compounds in the leaf and flower of Tithonia diversifolia (Hemsl) Gray. J Nat Med 61:63-66

Mosman,T., 1983. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay. J Immunol Methods 65,55-63

Nooter K, 1997, The role of P53 in resistance to cytotoxic therapy of human cancer. South West Cancer News. 4, 12-13

Nooter K, Burger H, Schenk P, Stoter G, 1999, Molecular mechanisms of drug resistance and sensitivity. Oncological Research at the Erasmus University Rotterdam-University Hospital Rotterdam, 39-40

Obafemi,C.A.,Sulaimon,T.O., Akinpelu, D.A., dan Olugbade, T.A., 2006. Antimicrobial activity of extract and a germacranolidetype sesquiterpene lactone from Tithonia diversifolia leaf extract. African Journal of Biotechnology. Vol 5 (12),pp 1254-1258

Parkin,D.M., Bray,F., Ferlay,J., dan Pisani,P., 2005. Global cancer statistics, 2002. CA Cancer J Clin 55: 74-108

Qomariyah,N., 2003. Herbal medicine: Pentingnya mengenal dan memahaminya. Mutiara Medika Jurnal Kedokteran Kesehatan 1(2):38-41

Tona,L., Kambu,K., Mesia,K., Cimanga,K., Apers,S., De Brynne,T., Pieters,L., Totte,J., dan Vlietinck,A.J., 1999. Biological screening of traditional preparation from some medicinal plants used as antidiarrhoeal in Kinshsa Kongo. Phytomedicine 6(1): 59-66

World Health Organization, 2002. Cervical Cancer Screening in Developing Countries: Report of a WHO Consultation. WHO, Geneva



DOI: https://doi.org/10.22146/tradmedj.7757

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