Sari Haryanti
Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional, Tawangmangu

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Ursolic Acid Enhances Doxorubicin Cytotoxicity on MCF-7 Cells Mediated by G2/M Arrest Arifin, Ibrahim; Hermawan, Adam; Ikawati, Muthi; Haryanti, Sari; Anindyajati, .; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 3, No 3 (2012)
Publisher : Indonesian Research Gateway

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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 Hedyotis corymbosa L. Increases Cytotoxic Activity of Doxorubicin on MCF-7 Breast Cancer Cell Haryanti, Sari; Junedi, Sendy; Meiyanto, Edy
Indonesian Journal of Biotechnology Vol 14, No 1 (2009)
Publisher : Universitas Gadjah Mada

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Abstract

Hedyotis corymbosa L. with ursolic acid as the main compound is one of the plants that has been used for traditional medicine including to cure breast cancer disease. The aim of this research is to examine the cytotoxic activity of rumput mutiara herb ethanolic extract (ERM) and its effect in combination with doxorubicin against MCF-7 breast cancer cell line as cell model of doxorubicin resistance. Hedyotis corymbosa L. herb powder extraction was done by maceration using ethanol 96% then the extract is detected for ursolic acid content. Cell viability assay of ERM, doxorubicin and  the combination of ERM and doxorubicin treatments were carried out by MTT assay to determine IC50 and CI (Combination Index). Cell cycle distribution was determined by flowcytometry. Apoptosis assay was performed by ethidum bromide-acridine orange DNA staining method. Investigation on Bcl-2 expression was determined by immunocytochemistry method. Thin Layer Chromatography of ERM had similar Rf with ursolic acid standard: 0,6. ERM and doxorubicin inhibited cell growth against MCF-7 with IC50  of 77 µg/mL and 349 nM (0,19 µg/mL) respectively. Combination of ERM and doxorubicin showed synergistic effect (CI 0.66-0.99). Combination of 25 ìg/mL ERM- 200 nM doxorubicin induced apoptosis and decreased Bcl-2 expression but showed no cell accumulation on cell cycle. Doxorubicin induced high cell accumulation in G2/M phase, but ERM at the concentration of 25 ìg/mL had a low effect in G1 phase, and ERM IC50 did not induce cell accumulation otherwise apoptosis. These results concluded that the apoptosis mechanism of combination doxorubicin-ERM is mediated by cell cycle arrest and non cell cycle arrest. Therefore ERM has a potential activity to be developed as co-chemotherapeutic agent.  
Aktivitas sitotoksik ekstrak air dan etanolik kulit manggis (Garcinia mangostana Linn.) pada beberapa model sel kanker Haryanti, Sari; Widayanti, Elok; Widiyastuti, Yuli
Jurnal Tumbuhan Obat Indonesia Vol 10, No 1 (2017): Jurnal Tumbuhan Obat Indonesia
Publisher : Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/jtoi.v10i1.6742.1-9

Abstract

Indonesia merupakan salah satu surga di Asia sebagai tempat tumbuh tanaman tropis dengan keunikan morfologi, rasa, dan khasiat biologis. Salah satunya adalah buah manggis (Garcinia mangostana Linn.), yang lebih dikenal dengan “ratu buah”. Rebusan kulit manggis digunakan oleh masyarakat di Indonesia untuk mengobati kanker. Data riset mengenai efikasi rebusan kulit manggis untuk kanker belum banyak dipublikasikan. Penelitian ini bertujuan untuk mengkaji aktivitas sitotoksik ekstrak air dan etanol dengan MTT assay pada sel HeLa, MCF-7, T47D, dan HepG2, serta karakterisasi golongan kimia menggunakan KLT. Ekstrak air (EAM) diperoleh dengan metode infusa, sedangkan ekstrak etanol 96% (EEM) dengan maserasi. Pengamatan sel setelah perlakuan kedua ekstrak memperlihatkan perubahan morfologi yang sesuai dengan kondisi apoptosis. Hasil MTT assay menunjukkan  perlakuan EEM dan EAM efektif terhadap sel kanker hati HepG2 dengan IC50 berturut-turut 96,1 and 87,3 μg/mL. Kedua ekstrak memiliki aktivitas sitotoksik lemah terhadap sel HeLa, MCF-7, dan T47D dengan rentang IC50 137-660 μg/ml. Berdasarkan arakterisasi KLT, kedua ekstrak mengandung senyawa terpenoid dan flavonoid. Dengan demikian, ekstrak kulit manggis potensial dikembangkan dalam pengobatan kanker hati.Indonesia is one of “the heaven place” in Asia for many tropical plants with interesting morphology, taste and biological activities. One of those plants is mangosteen (Garcinia mangostana Linn.), well recognized as “the queen of fruits”. Indonesian people have used mangosteen rinds to fight cancer nowadays. They usually prepared the rinds with boiling water. There is lack of data for this preparation efficacy for cancer treatment. This research aimed to evaluate cytotoxic activity of mangosteen-rinds aqueous and ethanolic extract on some human cancer cell line. This research also characterize the major compound of the extracts by TLC. Aqueous extract (EAM) was obtained by infusion, while ethanolic extract (EEM) by maceration method. MTT assay was done to determine viability of HeLa, MCF-7, T47D, and HepG2 cell line. The treatment of both extract against all cells caused morphological changes similar to apoptosis. The results of MTT assay revealed that EEM and EAM was effective against HepG2 with IC50 of 96,1 and 87,3 μg/mL respectively. Both had low activity against the other cell, with the IC50 137-660 μg/ml. The result of TLC characterization of both extracts revealed terpenoid and flavonoid as the major compound. This research suggest the mangosteen rind extract as potential candidate for hepatocarcinoma treatment.
The synergistic effect of doxorubicin and ethanolic extracts of Caesalpinia sappan L. wood and Ficus septica Burm. f. leaves on viability, cell cycle progression, and apoptosis induction of MCF­7 cells Haryanti, Sari; Pramono, Suwijiyo; Murwanti, Retno; Meiyanto, Edy
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.26105

Abstract

Caesalpinia sappan L. and Ficus septica Burm. f  known asa potential plant with wide variety of medicinal properties, including anticancer. Present study was aimed to explore cytotoxic effect ofsappan wood (ECS) and awar-awar leaves (EFS), and its combination with doxorubicin (dox) on MCF-7 cells focusing on cell cycle progression and apoptosis induction.The result of MTT assay showed that single treatment of ECS and dox performed cytotoxic effect with the IC50 value of 32 µg/mL and 6 µM respectively, while EFS performed low cytotoxic effect with the IC50 value of 282 µg/mL. The combination of ECS with EFS and doxorubicin showed synergistic cytotoxic effect. Flow cytometry analysis revealed that combination of ECS (16 µg/mL) with EFS (8 µg/mL) and doxorubicin (2 µM) induced apoptosis, and cell accumulation at sub-G1 and G2/M phases.Immunoblotting assay confirmed the apoptosis induction of this combination through increasing of cleavage of PARP-1. Based on these results, the synergistic cytotoxic effect of this combinationwas through G2/M phase accumulation and apoptosis inductionand potentially to be developed as co-chemotherapeutic agent.
Ethanolic Extract of Hedyotis corymbosa L. Increases Cytotoxic Activity of Doxorubicin on MCF-7 Breast Cancer Cell Haryanti, Sari; Junedi, Sendy; Meiyanto, Edy
Indonesian Journal of Biotechnology Vol 14, No 1 (2009)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.7809

Abstract

Hedyotis corymbosa L. with ursolic acid as the main compound is one of the plants that has been used for traditional medicine including to cure breast cancer disease. The aim of this research is to examine the cytotoxic activity of rumput mutiara herb ethanolic extract (ERM) and its effect in combination with doxorubicin against MCF-7 breast cancer cell line as cell model of doxorubicin resistance. Hedyotis corymbosa L. herb powder extraction was done by maceration using ethanol 96% then the extract is detected for ursolic acid content. Cell viability assay of ERM, doxorubicin and  the combination of ERM and doxorubicin treatments were carried out by MTT assay to determine IC50 and CI (Combination Index). Cell cycle distribution was determined by flowcytometry. Apoptosis assay was performed by ethidum bromide-acridine orange DNA staining method. Investigation on Bcl-2 expression was determined by immunocytochemistry method. Thin Layer Chromatography of ERM had similar Rf with ursolic acid standard: 0,6. ERM and doxorubicin inhibited cell growth against MCF-7 with IC50  of 77 µg/mL and 349 nM (0,19 µg/mL) respectively. Combination of ERM and doxorubicin showed synergistic effect (CI 0.66-0.99). Combination of 25 ìg/mL ERM- 200 nM doxorubicin induced apoptosis and decreased Bcl-2 expression but showed no cell accumulation on cell cycle. Doxorubicin induced high cell accumulation in G2/M phase, but ERM at the concentration of 25 ìg/mL had a low effect in G1 phase, and ERM IC50 did not induce cell accumulation otherwise apoptosis. These results concluded that the apoptosis mechanism of combination doxorubicin-ERM is mediated by cell cycle arrest and non cell cycle arrest. Therefore ERM has a potential activity to be developed as co-chemotherapeutic agent.  
The aqueous extract of Gerrardanthus macrorhizus caudex enhanced doxorubicin activity in MCF-7 human breast cancer cells Haryanti, Sari; Widiyastuti, Yuli; Wahyono, Slamet
Indonesian Journal of Biotechnology Vol 23, No 1 (2018)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.32519

Abstract

Gerrardanthus macrorhizus (GM) caudex, is traditionally used in cancer therapy by the Tetun people in Belu District, East Nusa Tenggara Province, Indonesia, where it is known as “akar batu”. This study aimed to explore the cytotoxic effects of G. macrorhizus caudex aqueous extract, as well as its combination with doxorubicin, on MCF-7 cells. Also investigated were the possible mechanisms of interaction through cell cycle progression and apoptosis induction. Single treatments of 5–320 µg/mL of the extract showed morphological alterations in MCF-7 cells, but did not show any cytotoxic effect. Combining the extract with doxorubicin resulted in a synergistic cytotoxic effect. Doxorubicin concentrations equivalent to 1/12, 1/8, and 1/5 fold of the IC50 combined with 20 µg/mL decreased viability to 48%. We then explored the combination effect of doxorubicin 0.4 µM with GM 5 and 20 µg/mL using a flow cytometer. A low concentration of the extract (5 µg/mL) combined with 0.4 µM of doxorubicin resulted in slight cell cycle modulation by G1, G2M arrested and apoptosis induction. The combination of doxorubicin and a higher concentration of the extract (20 µg/mL) did not show cell cycle modulation, and led to necrosis. Therefore, G. macrorhizus caudex at low concentrations has the potential to be developed further as a co-chemotherapeutic agent.
Evaluation of The Genotoxicity of Three Food Additives using CHO-K1 Cells under in vitro Micronucleus Flow Cytometry Assay Lestari, Beni; Novitasari, Dhania; Putri, Herwandhani; Haryanti, Sari; Sasmito, Ediati; Meiyanto, Edy
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/indonesianjcanchemoprev8iss2pp74-80

Abstract

Exposure of genotoxic substances come from various sources such as food additives. The aim of this study is to evaluate the genotoxicity of food additives in CHO-K1 cells by micronucleus test flow cytometry. The food additives: sodium saccharine (SS), monosodium glutamate (MSG), and sodium benzoate (SB) were assessed by in vitro cytotoxicity and genotoxicity using Chinese Hamster Ovary-K1 (CHO-K1) cells. The cytotoxic effect of those compounds was evaluated by MTT Assay on CHO-K1 Cells. The genotoxic evaluation was observed by in vitro micronucleus test by flowcytometry with double staining method. The results showed that the three compounds did not perform cytotoxic effect, increased the frequency of micronucleus, and changed the cell cycle profiles. In general, these studies obtained that none of three food additives showed cytotoxic and genotoxic effect on CHO-K1 cells. Micronucleus test using flow cytometry is suitable for this purpose study.Key words : food additives, genotoxic, cytotoxic, micronucleus
Different 4T1 Cells Migration under Caesalpinia sappan L. and Ficus septica Burm.f Ethanolic Extracts Haryanti, Sari; Murwanti, Retno; Putri, Herwandhani; Ilmawati, Gagas Pradani Nur; Pramono, Suwijiyo; Meiyanto, Edy
Indonesian Journal of Cancer Chemoprevention Vol 8, No 1 (2017)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev8iss1pp21-26

Abstract

Secang (C. sappan L.) and awar (F. septica Burm.f) are known of Indonesian traditional medicine that highly consumed throughout centuries in order to cure various diseases. Recently, researchers also concern about its effects as anti-cancer on various cell types. This study was conducted to understand the ethanolic extract of C. sappan L. (ECS) and F. septica Burm.f (EFS) effects on 4T1 cells migration at various concentrations. Firstly, we examine cell proliferation profile with MTT assay under treatment with the extracts and obtained the IC50 value of ECS (20 μg/mL) and EFS (15 μg/mL). Subsequent assay conducted was to examine the cells migration under low concentration resulting in the migration inhibitory effect of both EFS and ECS with different intensity.  EFS performed better migration inhibitory effect than ECS. Finally, we conducted gelatin zymography and western blot and revealed that the migration inhibitory effect of EFS may correlate to the lowering of protein expression of MMP9 and Rac-1 after 24 hours of treatment. We conclude that both extracts are potential to be developed as anticancer agent and EFS is more potent for anti-metastasis.Keywords: C. sappan L., F. septica Burm.f, 4T1, anti-migration
Ursolic Acid Enhances Doxorubicin Cytotoxicity on MCF-7 Cells Mediated by G2/M Arrest Arifin, Ibrahim; Hermawan, Adam; Ikawati, Muthi'; Haryanti, Sari; Anindyajati, Anindyajati; Meiyanto, Edy
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
Selectivity Index of Alpinia galanga Extract and 1’-Acetoxychavicol Acetate on Cancer Cell Lines Da'i, Muhammad; Meilinasary, Khairunnisa Azani; Suhendi, Andi; Haryanti, Sari
Indonesian Journal of Cancer Chemoprevention Vol 10, No 2 (2019)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev10iss2pp95-100

Abstract

Previous research stated that galangal (Alpinia galanga) extract has a potential as cytotoxic agent with active compound of 1?-Acetoxychavicol Acetate (ACA). The objective of this study was to determine the selectivity of ethanol extract, ethyl acetate fraction, and methanol fraction of of galangal, and ACA on cancer cell lines. Cytotoxic activity was carried out using the MTT method on T47D breast cancer, WiDr colon cancer, HeLa cervical cancer, and Vero normal cell lines. The results showed that galangal ethanol extract and its fractions had selectivity index equal to or less than 2 on cancer cells. Meanwhile, ACA had selectivity index more than 3 on T47D cell and HeLa cell. ACA showed a strong cytotoxic activity against cancer cells T47D, HeLa, and WiDr with IC50 values of 3.14, 7.26, and 12.49 ?g/ml, respectively. Based on data, it could be concluded that ACA was the most selective to inhibit T47D cell with a selectivity index of 6.6.Keywords: 1?-Acetoxychavicol acetate, galangal (Alpinia galanga), selective index, cytotoxic