Antiproliferative activity of Ethanolic Extract of Kembang Bulan (Tithonia diversifolia) Leaf on HeLa Cervical Cancer Cell Line

Endah Puspitasari, Nuri Nuri, Indah Yulia Ningsih, Bawon Triatmoko, Dewi Dianasari


Tithonia diversifolia has been showed to be cytotoxic and antiproliferative on colon cancer, glioblastoma, hepatoma, kidney cancer, breast cancer, lung cancer, melanoma, leukemia, ovary cancer, prostate cancer, and stomach cancer cell lines, but not on cervical cancer cells yet. Our research aimed to determine the cytotoxicity and antiproliferative activity of T.diversifolia leaf ethanolic extract on HeLa cervical cancer cell line. The cytotoxicity and the antiproferative activity assay were done using MTT method for 24 h for cytotoxic assay; and series of 24, 48, and 72 h for antiproliferative assay. The cytotoxic activity was analyzed using IC50, while the antiproliferative assay was analyzed based on the proliferation kinetics. All assays were done in triplicate. T.diversifolia leaf ethanolic extract exhibited strong cytotoxic activity on HeLa cervical cancer cell lines with the IC50 of 97.839±10.120 μg/mL. The cytotoxic activity was dose dependent. Based on the proliferation assay, the antiproliferative activity was stronger as the incubation time and the dose increases. T.diversifolia leaf ethanolic extract showed strong cytotoxic and antiproliferative activity on HeLa cervical cancer cell lines.

Keywords: T.diversifolia leaf ethanolic extract, cytotoxicity assay, antiproliferative assay, HeLa cervical cancer cells.

Full Text:



Ambrósio, S.R., Oki, Y., Heleno, V.C.G., Chaves, J.S., Nascimento, P.G.B.D., Lichston, J.E., et al., 2008, Constituents of glandular trichomes of Tithonia diversifolia: Relationships to herbivory and antifeedant activity, Phytochemistry, 69, 2052–2060. CrossRef

Calderón., Á.I., Vázquez, Y., Solís, P.N., Caballero-George, C., Zacchino, S., Gimenez, A., et al., 2006, Screening of Latin American plants for cytotoxic activity, Pharm. Biol., 44, 130–140. CrossRef

Centers for Disease Control and Prevention, Division of Cancer Prevention and Control, 2021, Side Effects of Cancer Treatment, Website,, accessed on Nov 22nd, 2021.

Chagas-Paula, D.A., Oliveira, R.B., Rocha, B.A., and Da Costa, F.B., 2012, Ethnobotany, chemistry, and biological activities of the genus Tithonia (Asteraceae), Chem. Biodivers., 9, 210–235. CrossRef

Chiang, L.C., Cheng, H.Y., Chen, C.C., and Lin, C.C., 2004, In vitro anti-leukemic and antiviral activities of traditionally used medicinal plants in Taiwan, Am. J. Chin. Med., 32, 695–704. CrossRef

Cragg, G.M., and Pezzuto, J.M., 2016, Natural products as a vital source for the discovery of cancer chemotherapeutic and chemopreventive agents, Med Princ Pract., 25 (suppl 2), 41-59. CrossRef

Doyle, A., and Griffiths, J.B., 2000, Cell and tissue culture for medical research, John Willey and Sons, Ltd., New York.

Fouche, G., Cragg, G.M., Pillay, P., Kolesnikova, N., Maharaj, V.J., and Senabe, J., 2008, In vitro anticancer screening of South African plants, J. Ethnopharmacol., 119, 455–461. CrossRef

Goffin, E., Ziemons, E., De Mol, P., De Madureira, M.D.C., Martins, A.P., Proença da Cunha, A., et al., 2002, In vitro antiplasmodial activity of Tithonia diversifolia and identification of its main active constituent: Tagitinin C, Planta Med., 68, 543–545. CrossRef

Gu, J.Q., Gills, J.J., Park, E.J., Mata-Greenwood, E., Hawthorne, M.E., Axelrod, F., et al., 2002, Sesquiterpenoids from Tithonia diversifolia with potential cancer chemopreventive activity, J. Nat. Prod., 65, 532–536. CrossRef

Wahyuningsih, M.S.H., Nugrahaningsih, D.A.A., and Budiyanto, A., 2019, Ethanolic Extract of Tithonia Diversifolia (Hemsley) a. Gray Inhibits Migration Activity and Decrease the Transforming Growth Factor-Beta1, Vegf Expression on Keloid Fibroblasts, Asian Journal of Pharmaceutical and Clinical Research, 12(1), 342. CrossRef

Kuroda, M., Yokosuka, A., Kobayashi, R., Jitsuno, M., Kando, H., Nosaka, K., et al., 2007, Sesquiterpenoids and flavonoids from the aerial parts of Tithonia diversifolia and their cytotoxic activity, Chem. Pharm. Bull., 55, 1240–1244. CrossRef

Lee, M.Y., Liao, M.H., Tsai, Y.N., Chiu, K.H., and Wen, H.C., 2011, Identification and anti-human glioblastoma activity of tagitinin C from Tithonia diversifolia methanolic extract, J. Agric. Food Chem., 59, 2347– 2355. CrossRef

Liao, M.-H., Lin, W.-C., Wen, H.-C., and Pu, H.-F., 2011, Tithonia diversifolia and its main active component tagitinin C induce survivin inhibition and G2/M arrest in human malignant glioblastoma cells, Fitoterapia, 82, 331–341. CrossRef

Liao, M.H., Tsai, Y.N., Yang, C.Y., Juang, C.L., Lee, M.Y., Chang, L.H., and Wen, H.C., 2013, Anti-human hepatoma Hep-G2 proliferative, apoptotic, and antimutagenic activity of tagitinin C from Tithonia diversifolia leaves, J. Nat. Med., 67, 98–106. CrossRef

Olayinka, B.U., Raiyemo, D.A., and Obukohwo, E., 2015, Phytochemical and proximate composition of Tithonia diversifolia (Hemsl.) A. Gray, Ann. Food Sci. Technol., 16, 195-200.

Orsomando, G., Agostinelli, S., Bramucci, M., Cappellacci, L., Damiano, S., Lupidi, G., et al., 2016, Mexican sunflower (Tithonia diversifolia, Asteraceae) volatile oil as a selective inhibitor of Staphylococcus aureus nicotinate mononucleotide adenylyltransferase (NadD), Ind. Crops Prod., 85, 181–189. CrossRef

Prayong, P., Barusrux, S., and Weerapreeyakul, N., 2008, Cytotoxic activity screening of some indigenous Thai plants, Fitoterapia, 79, 598-601. CrossRef

Ranti, I., Wahyuningsih, M.S.H., and Wirohadidjojo, Y.W., 2018, The antifibrotic effect of isolate tagitinin C from Tithonia diversifolia (Hemsley) A. gray on keloid fibroblast cell, Pan African Medical Journal, 30, 1–8.

Rocha, B.A., Pupo, M.T., Antonucci, G.A., Sampaio, S.V., Paiva, R.D.M.A., Said, S., et al., 2012, Microbial transformation of the sesquiterpene lactone tagitinin C by the fungus Aspergillus terreus, J. Ind. Microbiol. Biotechnol., 39, 1719–1724. CrossRef

Tagne, A.M., Marino, F., and Cosentino, M., 2018, Tithonia diversifolia (Hemsl.) A. Gray as a medicinal plant: a comprehensive review of its ethnopharmacology, phytochemistry, pharmacotoxicology and clinical relevance, J Ethnopharmacol, 220, 94-116. CrossRef

World Health Organization, 2018, Cancer Key Facts, Website,, accessed on March 20th, 2020.


Copyright (c) 2022 Endah Puspitasari, Nuri Nuri, Indah Yulia Ningsih, Bawon Triatmoko, Dewi Dianasari

Indexed by:




Indonesian Society for Cancer Chemoprevention