COMBINATION EFFECT OF ETHYLACETATE EXTRACTS OF PLECTRANTHUS AMBOINICUS (LOUR.) SPRENG. WITH DOXORUBICIN AGAINTS T47D BREAST CANCER CELLS
Keywords:Plectranthus amboinicus, doxorubicin, T47D, cell cycle, apoptosis
Objective: To investigated the growth-inhibiting and apoptosis mediating effect of Plectranthus amboinicus (Lour.) Spreng ethyl acetate extract (PAE) in combination therapy with doxorubicin against T47D cell lines, to analyzed the expression of cyclin D1 and COX-2 (cyclooxigenase-2) proteins.
Methods: The assays were performed in the study were cytotoxicity assay, cell cycle assay, apoptosis induction, and immunocytochemistry of T47D cells. The cytotoxicity effects were determined by using MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect on modulation of cell cycle and apoptosis was observed by flow cytometry assay in both single dose of PAE and its combination with doxorubicin. The expression of cyclin D1 and COX-2 proteins on T47D cell lines was identified by using immunocytochemistry.
Results: The result showed that the PAE 8 Âµg/ml had the synergistic effect with doxorubicin against T47D cells based on Combination Index analysis. The PAE induced apoptosis and cell accumulation at G1 phase. The combination of PAE 8 Âµg/ml with doxorubicin 1 Âµg/ml caused apoptosis induction same with its single treatment, but there was increasing in cell accumulation at G1 phase. Expression of cyclin D1 indicated that both single application and combination of PAE with doxorubicin could arrest the cell cycle of T47D cells. The expression of COX-2 showed that the combination could inhibit the metastasis of T47D breast cancer cells.
Conclusion: Based on the result that PAE could be a potential co-chemoterapeutic agent with doxorubicin on breast cancer cells.
Adina AB, Goenadi FA, Handoko FF, Nawangsari DA, Hermawan A, Jenie RI, et al. Combination of ethanolic extract of citrus aurantifolia peels with doxorubicin modulate cell cycle and increase apoptosis induction on MCF7 cells. Iran J Pharm Res 2014;13:919-26.
Rahmi F, Meiyanto E, Susidarti RA. Combination of doxorubicin and areca ethanolic extract induces apoptosis by increasing caspase-3 level on breast cancer (T47D) cells. Indonesian J Cancer Chemoprevention 2012;3:339-44.
Kumala S, Meiyanto E, Ikawati M, Jenie RI. Sinergisme fraksi butanol metabolit skunder kapang endofit 1.3.11 dengan doxorubicin dalam modulasi daur sel T47D dan MCF7. J Ilmu Kefarmasian Indonesia 2010;8:1-9.
Hasibuan PA, Rosidah Ilyas S, Nasution P. Antioxidant and cytotoxic activities of Plectranthus amboinicus, (Lour.) Spreng. Extracts. Int J Pharm Teach Practices 2013;4:755-8.
Rosidah and Hasibuan PA. Cytotoxic effect of n-Hexane, Ethylacetate and ethanol extracts of Plectranthus amboinicus, (Lour.) Spreng. on HeLa and vero cells lines. Int J PharmTech Res 2014;6:1806-9.
Anggraeni R, Hadisahputra S, Silalahi J, Satria D. Combinational effects of ethylacetate extract of Zanthoxylum acanthopodium DC. with Doxorubicin on T47D breast cancer cells. Int J PharmTech Res 2014;6:2032-5.
Satria D, Furqan M, Hadisahputra S, Rosidah. Combinational effects of ethylacetate extract of Picria fel-terrae Lour. and Doxorubicin on T47D breast cancer cells. Int J Pharm Pharm Sci 2015;7:73-6.
Hostanka K, Nisslein T, Freudenstein J, Reichling J, Saller R. Evaluation of cell death caaused by triterpene glycosides and phenolic substances form cimifigua racemosa extract in human MCF-7 breast cancer cells. Biol Pharm 2004;27:1970-5.
Kaewthawee N, Brimson S. The effect of ursolic acid on cytokine production via the MAPK pathways in leukemic T-Cells. EXCLI J 2013;12:102-14.
Shan J, Xuan Y, Zheng S, Dong Q, dan Zhang S. Ursolic acid inhibits proliferation and induces apoptosis of HT-29 colon cancer cells by inhibiting the EGFR/MAPK Pathway. J Zhejiang Univ Sci B 2009;10:674-68.
Neto CC. Ursolic acid and other pentacyclic triterpenoid: anticancer activities and occurence in berries. In: Stoner, Seeram. editors. Berries and Cancer Prevention. Vol 11. Dartmouth: Springer Press; 2011. p. 41-7.