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Edy Meiyanto
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada
Biochemistry, Genetics & Molecular Biology Medicine & Pharmacology

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Doxorubicin Induces Lamellipodia Formation and Cell Migration Nur Dina Amalina; Ika Putri Nurhayati; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 8, No 2 (2017)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev8iss2pp61-67

Abstract

Breast cancer is the main cause of cancer death among women, especially breast cancer metastasis. Metastasis process begins with the ability of cell cancer invasion. Doxorubicin, a antracycline chemotheraphy, is known to induce TGFβ1, thus promote invasion. The aim of this study is to optimize doxorubicin doses to induce lamellipodia formation in 4T1 and MCF-7/HER2 cells. Lamellipodia formation was observed by morphological changes using microscope inverted. The effect of doxorubicin on cell viability was analyzed using MTT assay. Rac1 expression after doxorubicin exposure was determined by western blotting. Lamellipodia formation was observed by morphological change of the cell at the dose 10, 25, 50 and 100 nM. Doxorubicin at the dose of 10 nM could induced lamellipodia formation without affect cell viability in both 4T1 and MCF-7/HER2 cells.  Doxorubicin induced cell cycle arrest at G2/M phase at all doses. Doxorubicin 10 nM also decrease Rac1 expression compared to control.Key words: Doxorubicin, lamellipodia, Rac1, migration.
Reveal Cytotoxicity and Antigenotoxicity of Piper nigrum L. Ethanolic Extract and its Combination with Doxorubicin on CHO-K1 Cells Nur Fitra Sari; Beni Lestari; Dian Saputri; Anisa Fauzia Ahsani; Ragil Anang Santoso; Ediati Sasmito; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 8, No 3 (2017)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev8iss3pp110-119

Abstract

Black pepper (Piper nigrum L.), one of the most popular Indonesian spices has been reported to possess various therapeutic effects. The aim of this study is to evaluate the cytotoxicity and antigenotoxicity of black pepper ethanolic extract (BPE) and its combination with doxorubicin (Dox) on CHO-K1 cells. Based on thin layer chromatographyanalysis, BPE contained piperine.Under MTT assay, BPE showed cytotoxic effect with the IC50 value of 68 μg/mL and performed synergism in combination with Dox. In vitro micronucleus test using Giemsa staining revealed that BPE did not cause morphological changes qualitatively on CHO-K1 cells at concentration of 8.5 μg/mL, whereas using flow cytometry analysis showed that BPE could decrease the number of micronucleus (MN) formation induced by doxorubicin. In addition, BPE reduced the ROS level on the CHO-K1 cells which observed by reactive oxygen species (ROS) intracellular assay. The decrease in ROS level indicated that the antioxidant activity of BPE contribute to the antigenotoxicity. Furthermore, molecular docking performed that piperine interacted with DNA Topoisomerase II with docking score of -80.68. Overall,BPE performed cytotoxic effect in single treatment, increased the cytotoxicity and reduced the genotoxicity of doxorubicin. Thus, BPE has potential to be developed further as co-chemotherapeutic and antigenotoxic agent.Keywords: Cytotoxic, genotoxic, Piper nigrum L., CHO-K1, micronucleus
Pentagamavunon-0 (PGV-0) Enhance Cytotoxic Effect of Doxorubicin through Increasing of Apoptosis, Senescence and ROS Level on Triple Negative Breast Cancer 4T1 Ismanurrahman Hadi; Riris Istighfari Jenie; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 11, No 1 (2020)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev11iss1pp7-15

Abstract

TNBC, one of the sub type of breast cancers was widely known with high tumorigenic and poor prognosis than others. The development of combination agent (co-chemotherapy) with doxorubicin for chemotherapy of TNBC were carried out to decrease doxorubicin side effect and resistance in cancer. This present study aims to explore the co-chemotherapeutic properties of PGV-0 and investigate induction of doxorubicin on apoptosis, senescence and ROS against TNBC. 4T1 Cell line were used as a TNBC in vitro model. Cytotoxic measurement was performed using MTT assay resulting in IC50 values of 52 μM. Meanwhile, the combination of doxorubicin and PGV-0 showed synergistic effect which decreased cell viability of 4T1 better than single treatment of doxorubicin. Apoptosis analysis was performed using annexin V/PI assay indicated that the combination treatment of PGV-0 and doxorubicin increased apoptosis evidence. Senescence detection was carried out using senescence-associated-β galactosidase (SA-β-gal) assay. The results showed that a single treatment of PGV-0 induced cellular senescence and increased senescence cells in combination treatment. Moreover, DCFDA staining showed that PGV-0 increased ROS level at single treatment, whereas combination treatment increased ROS intracellular compared to the positive control of doxorubicin. Based on these results, PGV-0 has potential as a co-chemotherapeutic candidate on TNBC.Keyword: 4T1, PGV-0, Co-chemotherapy, Cytotoxic, Senescence, Apoptosis, ROS
Fingerroot (Boesenbergia pandurata) Extract Inhibits Proliferation and Migration of 4T1 Metastatic Breast Cancer Cells Marsya Yonna Nurrachma; Gergorius Gena Maran; Nindya Budiana Putri; Yuni Fajar Esti; Adam Hermawan; Edy Meiyanto; Riris Istighfari Jenie
Indonesian Journal of Cancer Chemoprevention Vol 11, No 3 (2020)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev11iss3pp103-114

Abstract

Fingerroot (Boesenbergia pandurata) is an Indonesian herb, with anti-proliferation and anti-migratory effects against several cancer cells. This study aims to investigate the anticancer property of Fingerroot Extract (FE) in combination with doxorubicin (Dox) against 4T1, a metastatic breast cancer cell lines. FE was prepared by 96% ethanol maceration and characterized by thin-layer chromatography analysis. FE was subjected to a cytotoxicity test with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay alone or in combination with 10 nM Dox against 4T1 cells. Cytotoxic effect was then confirmed by measure reactive oxygen species (ROS) intracellular level using 2’,7’-dichloroflourescin diacetate (DCFDA)-staining flow cytometry-based assay. The anti-migratory effect was observed using scratch wound healing assay and gelatin zymography to investigate matrix metalloproteinase (MMP)-9 expression. FE showed a cytotoxic effect with an inhibitory concentration 50 (IC50) value of 25.5±3.9 μg/mL and performed an improved effect in combination with 10 nM Dox. A single treatment of FE decreased ROS intracellular level, while in combination with Dox, FE increased the ROS intracellular level. Further, at 42 h observation, FE and its combination with Dox inhibited the migration of 4T1 cells with % closure of 82.6 and 82.5, respectively, correlates with a significant decrease of MMP-9 expression. Overall, FE performs a cytotoxic activity and anti-migration activity on 4T1 breast cancer cells.Keywords: Boesenbergia pandurata, cytotoxic, ROS, anti-migration, 4T1 
The Doxorubicin-Induced G2/M Arrest in Breast Cancer Cells Modulated by Natural Compounds Naringenin and Hesperidin Sendy Junedi; Adam Hermawan; Aditya Fitriasari; Agustina Setiawati; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 12, No 2 (2021)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev12iss2pp83-89

Abstract

Doxorubicin as the common drug for breast cancer has been widely proposed to use in combine with a natural compound in order to overcome its side effects such as cardiotoxicity and resistance. Previously, we reported that naringenin and hesperidin, the abundant flavanons in citrus fruit peel, increased cytotoxic and apoptosis activities of doxorubicin in doxorubicin resistant breast cancer cells (T47D and MCF-7 cells). Since doxorubicin arrests G2/M phase in most cancer cells, both flavanons are speculated to affect the similar phase in breast cancer cells. Cell cycle distributions were determined by flowcytometry using propidium iodide (PI) to stain DNA of the cells. Combination of naringenin or hesperidin with doxorubicin increased accumulation of T47D cells in G2/M phase, while in MCF-7 cells, accumulated cells in G2/M phase were decreased, accompanying with slightly increased in G1 phase. Naringenin itself had no effect on cell cycle of both cells. Whereas, hesperidin arrested G2/M and G1 phases in T47D and MCF-7 cells, respectively. The different effect of naringenin and hesperidin in T47D and MCF-7 cells is most likely caused by difference of p53 status. In p53 mutant, T47D cells, naringenin and hesperidin supported mechanism of doxorubicin to arrest at G2/M that to be considered via p53-independent pathway. Whereas, in p53 wild-type MCF-7 cells, naringenin and hesperidin decreased G2/M arrest, suggesting that both flavanons do not utilize cell cycle arrest for their anticancer activity with doxorucibin. This study revealed that potential co-chemoterapeutic agents, naringenin and hesperidin distinctly modulated cell cycle arrest induced by doxorubicin according to the characteristic of breast cancer cells.Keywords: naringenin, hesperidin, doxorubicin, cell cycle, breast cancer cells.
Growth Inhibitory Property of Pentagamavunone-0 (PGV-0) on 4T1 Cells under Stress Condition : 2D and 3D Culture Model Haruma Anggraini Muflikhasari; Riris Istighfari Jenie; Ratna Asmah Susidarti; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 10, No 3 (2019)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev10iss3pp149-158

Abstract

Pentagamavunone-0 (PGV-0), one of the curcumin analogue, is reported to have a cytotoxic effect on various cancer cells. This study aimed to explore the growth inhibitory effects of PGV-0 against highly-metastatic breast cancer, 4T1 cells under stress condition covering 2D and 3D speroid cytotoxic, anti-migration, and suppression of MMP-9. PGV-0 showed cytotoxic effects on 2D and 3D 4T1 cells with IC50value of 49 μM and 26 μM, respectively. In addition, PGV-0 performed anti-migratory effect. The single treatment at 25 μM PGV-0 and 50 μM showed inhibitory effect on cell migration by 54% and 51% respectively. whilst, the combination of PGV-0 at the concentration of 25 μM and 50 μM with doxorubicin significantly inhibited cell migration by 41% and 38%,  respectively. The gelatin zymography assay showed that PGV-0 decreased MMP-9 expression both in a single treatment and in combination with doxorubicin. In conclusion,  PGV-0 is potential to be developed as anti-tumorigenesis agent on highly-metastatic breast cancers.Keywords: Pentagamavunone-0 (PGV-0), anti-migration, MMP-9, 4T1 cells, spheroid
Ursolic Acid Enhances Doxorubicin Cytotoxicity on MCF-7 Cells Mediated by G2/M Arrest Ibrahim Arifin; Adam Hermawan; Muthi' Ikawati; Sari Haryanti; Anindyajati Anindyajati; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 3, No 3 (2012)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev3iss3pp410-418

Abstract

Ursolic acid has been widely known to possess biological activity against numerous tumor cell lines. Previous studies revealed its cytotoxicity on several cancer cells in vitro by either inducing apoptosis or cell cycle modulation. This study was conducted to investigate ursolic acid’s cytotoxicity solely and in combination with a chemotherapeutic agent, doxorubicin, on MCF-7 breast cancer cells, followed by observation on its mechanism. Cytotoxicity of single and combinational treatment of ursolic acid and doxorubicin on MCF-7 breast cancer cells were conducted by using MTT assay. Single treatment was then evaluated by determining IC50 value, while combinational treatment was evaluated by analyzing cell viability and evaluating combination index (CI). To explore the mechanism underlying cytotoxic effect on respected cells, further analysis on cell cycle profile of single and combinational treatment was conducted by flow cytometry. Twenty four hours treatment of ursolic acid inhibited MCF-7 cells’ growth with IC50 value of 37 µM, while combinational treatment showed that several concentration combinations of ursolic acid and doxorubicin exhibited synergism of cytotoxic activity on MCF-7 cells, giving optimum CI value of 0.54. Flow cytometric analysis showed that combinational treatment induced G2/M arrest in MCF-7 cells. These results show that ursolic acid is promising to be developed as either single chemopreventive agent, or as doxorubicin’s co-chemotherapeutic agent in breast cancer treatment. Observation on the selectivity as part of safety aspect together with in silico, in vitro, and in vivo study on its molecular mechanism should be conducted.Keywords: ursolic acid, doxorubicin,co-chemotherapeutic agent, breast cancer, cell cycle
Ethanolic Extract of Secang (Caesalpinia sappan L.) Wood Performs as Chemosensitizing Agent Through Apoptotic Induction on Breast Cancer MCF-7 Cells Rahmi Khamsita; Adam Hermawan; Dyaningtyas Dewi Pamungkas Putri; Edy Meiyanto
Indonesian Journal of Cancer Chemoprevention Vol 3, No 3 (2012)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev3iss3pp444-449

Abstract

Resistance to chemotherapy is believed to cause treatment failure of the patient cancer. Secang (Caesalpinia sappan L.) has been proven to possess anticancer activity on some cancer cell lines. The aimed of this study to develop ethanolic extract of secang wood (EES) as chemosensitizing agent through apoptotic induction on breast cancer MCF-7 cells. Extraction of secang was done by using maceration with 70 % ethanol. Single and combinatorial treatment of EES and doxorubicin on MCF-7 breast cancer cells were analyzed by using MTT assay to determine the IC50 value and combination index (CI) to evaluate the combinatorial effect. Apoptosis was analyzed with flowcytometry (annexin V).  EES showed a dose-dependent cytotoxicity (IC50 value of 37 µg/ml), while combinatorial treatment showed that 7 concentrations was found to be synergist with doxorubicin on MCF-7 cells. Combinatorial treatment also triggered apoptotic instead of single treatment. Based on this result, we conclude that ethanolic extract of secang wood is potential as chemosensitizing agent in breast cancer.Keywords: Caesalpinia sappan L, MCF-7 cells, doxorubicin, apoptosis.  
Co-Authors Adam Hermawan Aditya Fitriasari Agustina Setiawati Amalina, Nur Dina Anindyajati Anindyajati Anisa Fauzia Ahsani Beni Lestari Dian Saputri Dyaningtyas Dewi Putri Pamungkas Ediati Sasmito Gergorius Gena Maran Haruma Anggraini Muflikhasari Ibrahim Arifin Ika Putri Nurhayati Ismanurrahman Hadi Marsya Yonna Nurrachma Muthi Ikawati Nindya Budiana Putri Nur Fitra Sari Ragil Anang Santoso Rahmi Khamsita Ratna Asmah Susidarti Riris Istighfari Jenie Sari Haryanti Sendy Junedi Yuni Fajar Esti