Indonesian Journal of Cancer Chemoprevention
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Cytotoxic Activity on MCF-7 Cells and in Silico Study of Sapogenin Steroids From Trigonella foenum-graecum L.
Get PDFTrigonella foenum-graecum (TFG) is one of medicinal plants containing several steroidal sapogenins, such as diosgenin, yamogenin, gitogenin, tigogenin and trigoneoside, and also alkaloid trigonellin and some flavonoids such as vitexin, isovitexin, orientin, isoorientin, which has many activities, such as antidiabetic, estrogenic, and also anticancer. As phytoestrogen, TFG was predicted to have potency as Selective Estrogen Receptor Modulators (SERMs) which is used for hormonal-dependent breast cancer treatment. This experiment was carried out to investigate interaction of some sapogenin steroids and flavonoids in TFG to estrogen receptor alpha (ERα) and its activity to breast cancer cell line as confirmation. In silico prediction was carried out to investigate their estrogenic activity by analyzing their binding affinity to ERα using AutoDock Vina program. In vitro activity confirmation of TFG extract and its fractions were carried out using MTT assay on Erα-positive human breast cancer cell line, MCF-7. Results showed that free binding energies of diosgenin and yamogenin were -6.4 kcal/mol, estradiol was -6.0 kcal/mol, and tamoxifen was -5.1 kcal/mol. While cytotoxicity assay showed that ethyl acetate fraction gave the lowest IC50 of 41.81 ppm, with total steroid content of 20.03 ppm. From these results, we can conclude that diosgenin and yamogenin have greater binding affinity to ERα comparing to estradiol and tamoxifen. In vitro assay confirmation showed that ethyl acetate fraction has cytotoxic effect on MCF-7 cells.Keywords: Trigonella foenum-graecum, sapogenin steroids, MCF-7, estrogen receptor alpha, binding affinit
Cytotoxic and Apoptotic Activity on Mcf7 Cell from Ethanolic Extract of Trigonella foenum-graecum L., Aglaia elliptica Blume. and Foeniculum vulgare Mill.
Get PDFIndonesian medicinal herbs have so many biological activity, including anticancer activity. This assay aims to evaluate three Indonesian medicinal plants (Trigonella foenum-graecum L. (TFG), Aglaia elliptica Blume. (AE) and Foeniculum vulgare Mill. (FV)) which have potency in anticancer activity. The experiment was done by MTT assay to observe the cytotoxicity activity on breast cancer cell lines MCF7. Apoptotic potency was evaluated based on % population of MCF7 cell in subG0/G1 phase. The results showed that IC50 from ethanolic extract of TFG was 241.24 ppm, ethanolic extract of AE was 19.442 ppm and ethanolic extract of FV was 69.41 ppm. Apoptotic potency for TFG was 6.46%, AE was 11.94% and for FV was 8.73%. Thus, it is concluded that ethanolic extract of AE gives the lowest IC50 and the higher potency of apoptotic by significantly (p<0.05) increase % population of cells in subG0/G1 phase on 100ppm concentration.Keywords : Trigonella foenum-graecum L., Aglaia elliptica Blume. and Foeniculum vulgare Mill., MCF-7, Apoptotic, Cytotoxi
Combination Effect of N-Hexane Extract of Plectranthus amboinicus (Lour.) Spreng. with Doxorubicin Againts HeLa Cell Lines
Get PDFThe study was aimed to investigate the growth inhibiting effect of Plectranthus amboinicus, (Lour.) Spreng n-hexane extract (PAN) in combination with doxorubicin againts HeLa cell Lines, to observe the apoptotic induction and immunocytochemistry of HeLa cell Lines on cyclin D1, Bcl-2, and Cox-2 after treatment of PAN. The percentage viability of the cell were carried out by using MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of apoptosis was observed by flowcytometry assay in single dose of PAN. The expression of cyclin D1, Bcl2 and COX-2 proteins were investigated on HeLa cell lines by using immunocytochemistry. The result showed that PAN showed strong synergistic effect with doxorubicin againts HeLa cells based on Combination Index analysis. No significant apoptotic induction of PAN in HeLa cell lines, but the extract caused necrosis. The immunocytochemical study showed suppression of cyclin D1, Bcl2 and COX-2 expression on HeLa cell lines. The results concluded that PAN could be a potential co-chemoterapeutic agent with doxorubicin on cervix cancer cells but need to be explored further by its combination on specific molecular target.Keywords : Plectranthus amboinicus, (Lour.) Spreng., combination, HeLa cell line
Annona muricata's Leaves Ethanolic Extract Enhance p53 Expression in Rat Hepatic Cells Induced by Dimethylbenz(a)Anthracene (DMBA)
Get PDFAnnona muricata or soursop contains acetogenin which can induce apoptosis in vitro and has cytotoxic effect. The study has been done to examine soursop leaves extract’s potency in vivo especially to induce apoptosis in 7,12-Dimethylbenz[a]anthracene (DMBA)-induced Sprague Dawley rats. The induction process was done twice a week for five weeks. The soursop leaves extract were given in three dosages, that are 200, 400, and 800 mg/kgBW for seventeen days after two weeks DMBA induction. Data analysis showed that Annona muricata leaves ethanolic extract could enhance p53 pathway significantly with a dose independent manner in DMBA-induced rats.Keywords: Proapoptosis, Annona muricata leaves ethanolic extract, hepatom
Cytotoxic Activity and Apoptosis Induction of Ethanolic Extract of Pericarps of Mangosteen (Garcinia mangostana Linn.) on WiDr Cells and Interaction Study of Alpha-mangosteen to IKK and VEGF Based on Molecular Docking
Get PDFOne of the compounds found efficacious as an anti-proliferative on colon cancer is alpha-mangosteen, a xanthon compound that is abundant in pericarp of mangosteen. This study focused to evaluate anticancer activity of the ethanolic extract of pericarp of mangosteen (Garcinia mangostana Linn.) on WiDr colon cancer cells and to observe the affinity of alpha-mangosteen in inhibiting IKK and VEGF. Cytotoxic effect was tested by MTT assay and apoptosis induction was evaluated by double staining method on WiDr colon cancer cells, while interaction between alpha-mangosteen to the receptors was measured by molecular docking. The ethanolic extract was proven to have cytotoxic activity against WiDr colon cancer cells with IC50 of 25 µg/mL. In addition, the observation of apoptosis induction by double-staining method showed that apoptosis associated the cytotoxic effect of ethanolic extract of pericarp of mangosteen (EPM) on WiDr colon cancer cells. Molecular docking showed that active compounds in pericarp of mangosteen could compete with the native ligand of VEGF because the docking score of alpha-mangosteen was almost equal with native ligand. From these results we could be concluded that the cytotoxic effect of EPM to WiDr cells was mediated by cell apoptosis. This extract was potential to be developed as chemopreventive agent.Keywords : Garcinia mangostana Linn., cytotoxic effect, apoptosis, WiDr cell, molecular docking
Potency of Radical Scavenging Activity and Determination of Total Phenolic Content of Five Ethanolic Extract of Rhizome Zingiberaceae Family
Get PDFSeveral studies show that some plants and fruits could protect human body from free radical danger exposure. Zingiberaceae family has some chemical substances that have antiradical activity, such as phenolic compound and flavonoid. The aim of this study is to determine the correlation of total phenolic compound towards its antiradical activity. Antiradical activity assay was determined by DPPH method (1,1 difenil-2-pikrilhidrazil). The total phenolic compound is determined by spectrofotometry using Folin-Ciocalteu reagent and counted as GAE (Gallic Acid Equivalent). Based on this study, the total phenolic concentration of ethanolic extract of jahe (Zingiber officinale (L.) Rosc), kencur (Kaemferia galanga L.), lengkuas (Languas galanga), lempuyang gajah (Zingiber littorale) and lempuyang pahit (Zingiber americana) is 73.99 mg/g; 23.848 mg/g; 37.841 mg/g; 49.725 mg/g and 40.802 mg/g respectively. The radical scavenging activity (IC50) of jahe (27.38 µg/mL), lengkuas (93,27 µg/mL), lempuyang pahit (105,08 µg/mL), meanwhile lempuyang gajah and kencur show low activity of radical scavenging.Keywords: Zingiberaceae, total phenolic compound, antiradical, DPP
Ethanolic Extract of Secang (Caesalpinia sappan L.) Wood Performs as Chemosensitizing Agent Through Apoptotic Induction on Breast Cancer MCF-7 Cells
Get PDFResistance 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.
Ethanolic Extract of Papaya (Carica papaya) Leaf Exhibits Estrogenic Effects In Vivo and In Silico
Get PDFThe menopause women have the low level of estrogen in the body. The lack of estrogen changes physiological function in women’s body that affects in health condition. Carica papaya L. leaf contains flavonoid quercetin which exhibits estrogenic effect. The aim of this study is to determine the estrogenic effect of papaya leaves extract (PLE) in vivo, and in silico. Papaya leaves were extracted by ethanol 70% maceration. The in silico study were done by molecular docking between quersetin and Estrogen Receptor (ERα and ERβ) to obtain the docking score. Based on this study, docking score of quercetin was almost similar to the native ligand of ER. The in vivo study was done as follow: 36 female rats Sprague Dawley divided into six groups. The groups are shame-ovariectomized (S-OVX), control ovariectomized (OVX), CMC-Na control (OVX+CMC-Na), positive control (OVX+Estradiol), and the PLE treatment groups dose 750 mg/kgBW (OVX+750mg/kgBW) and dose 1000 mg/kgBW (OVX+1000 mg/kgBW). Administrations of PLE were done in three weeks orally, while estradiol was administrated intraperitonially. The mammae and uterine were sliced for analysis. Based on the study, the treatment of PLE increased the number of mammae lobules and uterine weight as well as estrogen does. In summary, PLE can be developed as a source of phytoestrogens.Keywords: Carica papaya L., phytoestrogen, estrogen receptor, mammae lobule, uterin
Ursolic Acid Enhances Doxorubicin Cytotoxicity on MCF-7 Cells Mediated by G2/M Arrest
Get PDFUrsolic 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 cycl
Secang (Caesalpinia sappan L.) Heartwood Ethanolic Extract Shows Activity as Doxorubicin Co-chemotherapeutic Agent by Apoptotis Induction on T47D Breast Cancer Cells
Get PDFDoxorubicin, primary chemoteurapeutic agent used for breast cancer treatment, is known to have various side effects included multi drug resistance (MDR) phenomenon. Therefore, exploration of co-chemotherapeutic agent is important to be conducted in order to prevent MDR. Secang (Caesalpinia sappan L.) which contains active compounds brazilin and brazilein, is proven to have activity as anticancer. The aim of this study is to determine the potency of Caesalpinia sappan L. ethanolic extract (CEE) as co-chemotherapeutic agent of doxorubicin and its mechanism through apoptosis induction on T47D breast cancer cells. Caesalpinia sappan L. heartwood powder was macerated with ethanol 70%. The cytotoxic effect of CEE alone and its combination with doxorubicin was analyzed using MTT assay. Apoptosis assay was done by flowcytometry-annexin V method. CEE showed cytotoxic activity on T47D cells with IC50 value of 35 µg/ml, while combinatorial test showed that all of combination doses of CEE and doxorubicin gave synergistic effect. Flowcytometry-annexin V assay proved that treatment of CEE induced apoptosis of doxorubicin. Based on these results, we conclude that Caesalpinia sappan L. heartwood ethanolic extract is potential to be developed as co-chemotherapeutic agent of doxorubicin.Keywords : Caesalpinia sappan L., doxorubicin, apoptosis, T47D cell