PENINGKATAN AKTIVITAS SITOTOKSIK DOKSORUBISIN OLEH NARINGENIN PADA SEL KANKER SERVIKS HeLa MELALUI MODULASI DAUR SEL DAN PEMACUAN APOPTO

Naringenin, an abundant flavanon in the peel of citrus fruits is reported to possess anti-proliferative effect in many cancer cells. Herein, we investigated the cytotoxic effect and apoptosis induction of naringenin in combination with doxorubicin on HeLa cells. The cytotoxicity assay of naringenin,...

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Bibliographic Details
Main Authors: , Larasati, , Prof. Dr. Edy Meiyanto, M.Si., Apt.
Format: Thesis
Published: [Yogyakarta] : Universitas Gadjah Mada 2011
Subjects:
ETD
Description
Summary:Naringenin, an abundant flavanon in the peel of citrus fruits is reported to possess anti-proliferative effect in many cancer cells. Herein, we investigated the cytotoxic effect and apoptosis induction of naringenin in combination with doxorubicin on HeLa cells. The cytotoxicity assay of naringenin, doxorubicin, and their combination were carried out by using MTT method. Cell viability and combination index were used as the parameters to evaluate combination effectiveness. Cell cycle distribution was determined by flow cytometry and then analyzed by using ModFit LT 3.0 program. Apoptosis assay was done using double staining method using Ethidium Bromide-Acridine Orange. Investigation on the expression of Bax and Bcl-2 were determined by immunocytochemistry method. Naringenin and doxorubicin showed cytotoxic effect on HeLa cells with their IC50 values of 195 uM and 1 µM, respectively. Treatment of naringenin combined with doxorubicin for 24 and 48 hours showed greater cytotoxicity compared with single treatment of doxorubicin. Based on CI values, combination naringenin 25 µM and doxorubicin 0,5 µM was the best synergistic effect. Single treatment of 0,5 µM doxorubicin for 24 hours in HeLa cells induced S-phase arrest. Although 100 µM naringenin did not affect HeLa cell cycle, its combination with 0,5 µM doxorubicin induced S-phase arrest with increased percentage of sub-G1 phase. In accordance with the flow cytometry results, the apoptosis assay of combination treatment indicated increase of apoptotic cells compared than single treatment. Naringenin, doxorubicin, and their combination also increased the expression of Bax and decreased the expression of Bcl-2. These results conclude that naringenin is a potential co-chemotherapy agent for cervical cancer due to its synergism with doxorubicin.