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All Journal Indonesian Journal of Cancer Chemoprevention
Larasati Larasati
Cancer Chemoprevention Research Center, Faculty of Pharmacy, Universitas Gadjah Mada
Biochemistry, Genetics & Molecular Biology Medicine & Pharmacology

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Hesperidin Increase Cytotoxic Activity of Doxorubicin on Hela Cell Line Through Cell Cycle Modulation and Apoptotis Induction Indri Kusharyanti; Larasati Larasati; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 2, No 2 (2011)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev2iss2pp267-273

Abstract

Combination of chemotherapeutic agent and chemopreventive agent is being a new approach in cancer treatment. This is aimed at enhancing the effectivity and also reducing drug resistance and adverse side effect of the chemotherapeutic agent. Hesperidin, a citrus flavonoid has reported to reduce the proliferation of many cancer cells. The objectives of this study were to investigate cytotoxic activities, cell cycle modulation and apoptosis induction of hesperidin and its combination with doxorubicin on Hela cell lines. MTT [3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide] assay was used to measure the growth inhibitory effect of hesperidin and its combination with doxorubicin on Hela cells. Cell cycle profile was determined by flowcytometry and the data obtained was analyzed by using ModFit LT 3.0 program. Apoptosis assay was done using double staining method using ethidium-bromide and acridine-orange. Hesperidin inhibited cell growth with IC50 48 μM, while the IC50 of doxorubicin was 1000 nM. Combination of 500 nM doxorubicin and 6 μM hesperidin showed strongest inhibitory effect toward Hela cells. Hesperidin of 24 µM accumulated HeLa cells at G1 phase, but its combination with 500 nM Doxorubicin gave G1 and S phase accumulation at 24 h incubation. Both of Hesperidin and Doxorubicin were capable of inducing apoptosis. In accordance of the apoptotic effect, hesperidin, doxorubicin and their combination decreased the expression Bcl-2 and increased the expression of Bax. According to this result, hesperidin has a potency to be developed as co-chemotherapeutic agent for cervical cancer.Keywords: Cochemotherapy, Hesperidin, Doxorubicin, Hela, MTT assay
Naringenin Enhances the Anti-Tumor Effect of Doxorubicin on HeLa Cervical Cancer Cells Through Cytotoxic Activity and Apoptosis Induction Larasati Larasati; Indri Kusharyanti; Adam Hermawan; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 2, No 3 (2011)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev2iss3pp325-333

Abstract

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 assay. Cell viability was used as the parameters to evaluate combination effectiveness. Cell cycle distribution was determined by flow cytometry and analyzed using ModFit LT 3.0 program. Apoptosic assay was done by 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 µM and 1 µM, respectively. Whereas combination of naringenin - doxorubicin showed greater cytotoxicity compared the single treatment of doxorubicin. The strongest cytotoxic activity was observed at a combination of 100 µM naringenin and 0,5 µM doxorubicin. Single treatment of 0,5 µM doxorubicin for 24 hours on HeLa cells induced S-phase arrest while 100 µM naringenin did not affect on HeLa cell cycle. The combination induced S-phase arrest with the increased of sub-G1 phase percentage. In accordance with the flow cytometry results, the double staining apoptosis assay results showed the increase of apoptotic cells. Naringenin, doxorubicin, and their combination also increased the expression of Bax and decreased the expression of Bcl-2. These results concluded that naringenin was a potential co-chemotherapy agent for cervical cancer due to its synergism with doxorubicin.Keywords: co-chemotherapy, naringenin, doxorubicin, HeLa cells, cytotoxicity, cell cycle, apoptosis
Leunca (Solanum nigrum L.) Herbs Ethanolic Extract Increase Cytotoxic Activity of Cisplatin on Hela Cervical Cancer Cells Raditya Prima Istiaji; Maya Fitria; Larasati Larasati; Fortunella Tjondro; Astrid Ayu Maruti; Erna Prawita Setyowati; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 1, No 1 (2010)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev1iss1pp32-37

Abstract

Cervical cancer is one of leading causes of cancer death in women in the developing countries. The use of cisplatin as chemotherapy agent in cervical cancer is known to cause side effects and also resistance for long-term uses. One of the strategies to prevent cervical cancer based on combination agents is being developed. Leunca (Solanum nigrum L.) has been revealed to inhibit growth of human cancer cells. Therefore, it can be used in combination with cisplatin to reduce those side effects and prevent the occurrence of cell resistance. Ethanolic extract of Leunca Herb (ELH) and cisplatin were tested their cytotoxic effect on HeLa cervical cancer cell by using MTT assay to determine IC50 value. The combinationss of cisplatin-ELH were tested to determine the combination index (CI value). The IC50 of ELH and cisplatin on HeLa cells were 227 µg/mL and 17 µM. rRespectively. Tthe study of combination resulted that almost all the index combinations were <0,9 showed the effect of synergism combination. The Ooptimum concentration of combination was 1/8 IC50 cisplatin–1/8 IC50 ELH. The results indicated that ELH had a potency to be combination agent to enhance the activity of cisplatin on HeLa cervical cancer cells. Therefore, further study on its molecular mechanism needs to be explored. Key words: Leunca (Solanum nigrum L.), cisplatin, cytotoxic, combination agent, HeLa cells
Co-Authors Adam Hermawan Astrid Ayu Maruti Edy Meiyanto Erna Prawita Setyowati Fortunella Tjondro Indri Kusharyanti Maya Fitria Raditya Prima Istiaji Ratna Asmah Susidarti