scholarly journals Enhancement of Cytotoxicity and Apoptosis Induction of Doxorubicin by Brazilein Containing Fraction of Secang (Caesalpinia sappan L.) on T47D Cells

Author(s):  
Rohmad Yudi Utomo ◽  
Annisa Novarina ◽  
Prisnu Tirtanirmala ◽  
Ria Fajarwati Kastian ◽  
Riris Istighfari Jenie

Combination chemotherapy (co-chemotherapy) is a recent strategy to reduce the toxicity effect and increase the effectivity of chemotherapeutic agent, such as Doxorubicin (Dox). Caesalpinia sappan L. are potential to be developed as co-chemoterapeutic agents due to its strong cytotoxicity toward several breast cancer cells. The purpose of this research is to observe the cytotoxicity of Brazilein containing fraction (BCF) in single and its combination with doxorubicin on T47D cells. BCF was obtained by fractionation using chloroform:ethyl acetate (40:60 v/v) as mobile phase. Molecular docking results showed that Brazilein and Brazilin interacted with Bcl-2 with different binding properties. Based on MTT assay, Dox and BCF performed potent cytotoxicity with IC50 value of 403 nM and 68 μg/mL, respectively. BCF increased the cytotoxicity of Dox and performed synergism with CI value <1 and decreased possible toxicity with DRI value>1. Under Annexin V PI staining Flowcytometry, BCF in single and its combination with doxorubicin induced apoptosis. In conclusion, single treatment of BCF and its combination with Dox performed cytotoxic effect and induced apoptosis on T47D cell lines.Keywords: Brazilein containing fraction, Doxorubicin, Co-chemoteraphy, Apoptosis, T47D cells

Author(s):  
Ika Nurzijah ◽  
Dyaningtyas Dewi Pamungkas Putri ◽  
Erlina Rivanti ◽  
Edy Meiyanto

Doxorubicin, 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 cells


2015 ◽  
Vol 12 (2) ◽  
pp. 55-59
Author(s):  
Edy Meiyanto

As chemotherapeutic backbone for breast cancer therapy, doxorubicin showed various side effects and induced resistancy of breast cancer cells. Development of targeted therapy on breast cancer focused on combinatorial therapy of doxorubicin and molecular targeted agents. PGV-0 and PGV-1, a curcumin analogue showed potency as co-chemotherapeutic agent with doxorubicin. Our previous study of PGV-0 and PGV-1 showed cytotoxic activity in T47D cells. Therefore, the aim of this research is to examine the synergistic effect of PGV-0, PGV-1 on the cytotoxic activity of doxorubicin through cell cycle modulation and apoptotic induction on MCF-7 breast cancer cell lines. The cytotoxic assay of PGV-0, PGV-1, doxorubicin, and their combination were carried out by using MTT assay. Cell cycle distribution and apoptosis were determined by flowcytometer FACS-Calibur and the flowcytometry data was analyzed using Cell Quest program. Single treatment of PGV-0, PGV-1 and doxorubicin showed cytotoxic effect on MCF-7 with cell viability IC50 value 50 µM, 6 µM and 350 nM respectively. Single treatment of Doxorubicin 175 nM induced G2/M arrest. Single treatment of PGV-0 5 µM induced G2/M arrest while in higher dose 12.5  µM, PGV-0 induced apoptosis. Combination of doxorubicin 175 nM and PGV-0 5 µM induced apoptosis. Combination of doxorubicin 175 nM and PGV-0 12.5 µM also increased apoptosis induction. Single treatment of PGV-1 0.6 µM induced G1 arrest while in higher dose 1.5  µM, PGV-1 induced apoptosis. Combination of doxorubicin 175 nM and PGV-1 0.6 µM induced apoptosis. Combination of doxorubicin 175 nM and PGV-0 1.5 µM also increased apoptosis induction. PGV-0 and PGV-1 are potential to be delevoped as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle modulation, but the molecular mechanism need to be explored detail.  Key words: PGV-0, PGV-1, doxorubicin, co-chemotherapy, breast cancer, cell cycle arrest, apoptosis


Author(s):  
Prisnu Tirtanirmala ◽  
Annisa Novarina ◽  
Rohmad Yudi Utomo ◽  
Raisatun Nisa Sugiyanto ◽  
Riris Istighfari Jenie ◽  
...  

Anticancer activity of secang’s heartwood (Caesalpinia sappan L.) is based on its main compound: brazilin and brazilein. Brazilin, brazilein, and other compounds such as caesalpiniaphenol can affect proteins that have a role in apoptosis. In this study, we observed cytotoxic activity of fraction containing brazilein (FCB) alone or in combination with chemotherapeutic agent, cisplatin and the ability of the combination to induce apoptosis in T47D breast cancer cell lines. Cytotoxicity assay was determined using MTT assay, whereas the detection apoptosis induction was conducted using flow cytometry using Annexin-V and propidium iodide. FCB and cisplatin showed cytotoxic effect on T47D cells with IC50 value of 68 µg/mL and 16 µM, respectively. Combination of FCB and cisplatin result synergistic combination at the concentration ratio of 1/2 IC50 with CI value of 0.66. Its combination also able to induce apoptosis on T47D cell population 13% larger than the single treatment. Based on this study, we conclude that FCB is able to enhance the cytotoxic effects of cisplatin by inducing apoptosis.Keywords:  Caesalpinia sappan L., cisplatin, apoptosis, breast cancer


Author(s):  
Inna Armandari ◽  
Kartika Dyah Palupi ◽  
Sofa Farida ◽  
Adam Hermawan ◽  
Ratna Asmah Susidarti ◽  
...  

Doxorubicin is one of chemotherapeutic agent widely used in breast cancer treatment, but in high dose doxorubicin gives negative side effect, including vomit, nausea, immune suppression, and cardiac toxicity. This toxicity hopefully could be reduced by combination chemotherapy using natural herbs such as ciplukan herb. This research was conducted to explore cytotoxic activity of single ciplukan herbs ethanolic extract and its combination with doxorubicin on T47D breast cancer cells. Cytotoxic activity of ciplukan herbs ethanolic extract only and its combination with doxorubicin were tested on T47D cells using MTT assay to obtain IC50 value and combination index (CI), respectively. Single extract showed cytotoxic activity on T47D cells with IC50 value of was 160 µg/ml. Thus, combination treatment from ciplukan herbs ethanolic extract and doxorubicin showed synergistic effect (CI<1,0). This effect was reached at concentration of ciplukan herbs ethanolic extract-doxorubicin 80 μg/ml- 2 nM, 80 μg/ml-4 nM, and 80 μg/ml-8 nM. This research indicated that ciplukan herbs ethanolic extract is potential to be applied as co-chemotherapeutic agent in breast cancer therapy.Key word : ciplukan herbs, doxorubicin, co-chemotherapy, T47D cells


Author(s):  
Poppy Anjelisa Zaitun Hasibuan ◽  
Rosidah ◽  
Pandapotan Nasution ◽  
Syafruddin Ilyas

The objectives of the study are to investigated the growth-inhibiting and apoptosis mediating effect of Plectranthus amboinicus (Lour.) Spreng n-hexane extract (PANE) on T47D cell lines. The assays were performed in the study were cytotoxicity assay and apoptosis induction of T47D cells. The cytotoxicity effects were determined by using MTT [3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The effect of apoptosis were observed by ethidium bromide-acrydine orange method. The growth of T47D was inhibited by treatment with PANE on concentrations 20; 40; dan 80 µg/mL. The cell death (apoptosis) induced by PANE was characterized by orange fluorescent on the IC50 concentration. These results concluded that the n-hexane extract of Plectranthus amboinicus, (Lour.) Spreng. inhibited the growth of T47D breast cancer cells in dose and time dependent manners. PANE also could induced apoptosis on T47D breast cancer cells, but the combination of PANE-doxorubicine did not show the synergistic effect. The results suggesting that PANE may be a potent agent for the chemoprevention of breast cancer.


2017 ◽  
Vol 21 (1) ◽  
pp. 29 ◽  
Author(s):  
Sari Haryanti ◽  
Suwijiyo Pramono ◽  
Retno Murwanti ◽  
Edy Meiyanto

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.


Author(s):  
Rollando Rollando

Objective: Ursolic acid was a compound found in Hedyotis corymbosa L., (HCoL) while berberine found in Tinospora crispa (TCa) which are proven to have cytotoxic effect to cancer cells. This research aims to review the effect of cisplatin, ethanolic extract of HCoL and TCa to the sensitivity increase on breast cancer cells, which will be confirmed through apoptosis induction and cell cycle modulation.Methods: The cytotoxic effect was tested using 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide assay on T47D cell using the IC50 parameter. The combination was tested by determining their combination index (CI) and cell viability. The combination effect of apoptosis induction and cell cycle modulation was observed using flow cytometry method.Results: The cytotoxic test result of the combination shows CI value of below 1 at the concentration of HCoL ethanolic extract as much as 1 μg/mL, TCa ethanolic extract as much as 6 μg/mL, and cisplatin as much as 2,5 μM. The combination of HCoL ethanolic extract, TCa ethanolic extract, and cisplatin results in phase S cell accumulation (29.98%) on breast cancer cell T47D and was able to induce apoptosis.Conclusion: The result proves that ethanolic extract of HCoL and TCa can be developed as a cochemotherapeutic agent with cisplatin to increase the effectivity of breast cancer treatment.


Author(s):  
Rochmah Kurnijasanti ◽  
Amaq Fadholly

Capsicum annum L. is a potential natural plant that have a lot of various pharmacological effects, including as anticancer agent. This study Aim to analyze Capsicum annum extract (CAE) on T47D cells. CAE (10, 20, 40, 60, 80µg/mL) treated on T47D cells to determined IC50 value by MTT assay. Apoptosis induction is also investigated through caspase-3 expressions (IC50, 2IC50). The present study showed that CAE suppress T47D cells proliferation with IC50 value of 75.81µg/mL. The caspase-3 expression on 2IC50 is higher (67.16%) than IC50 (52.16%). This result indicate that CAE has ability as anticancer agent by inhibiting cell growth and induce apoptosis through caspase-3 expression on T47D cells. Further study of CAE holds potential for novel therapies of cancer prevention and treatment.


2015 ◽  
Vol 2 (1) ◽  
pp. 64
Author(s):  
Mukhlish Jamal Musa Holle ◽  
Hestri Dyah Puspitasari ◽  
Andaru Satryo ◽  
Wahyu Dewi Astuti Ningrum ◽  
Digdo Sudigyo ◽  
...  

Breast cancer is one of cancer with high mortality. This cancer not only attacks women, but also men. Indonesia has many plants which potential as anticancer, such as orchids. Spathoglottis plicata is one of the orchid species that abundant in Indonesia and has a lot of antioxidant compounds which is guessed have anticancer properties. The objectives of this study were to study the cytotoxic activity and IC50 value of aquadest, ethanolic, and chloroform extracts of S. plicata’s pseudobulbs, leaves, and whole plants on T47D cells (breast cancer cells line) as well as cytotoxic activity of the specific fraction of the most toxic crude extract. S. plicata used in this study was obtained from Bungarinte nursery. Extractions were done by maceration method using aquadest, ethanol, and chloroform as the solvent. Cytotoxic test on T47D cells were done by MTT assay. The cytotoxic data were analyzed using one-way ANOVA followed by Tukey’s HSD test. The IC50 of each extracts were calculate by probit analysis. The lowest IC50 value among all extracts was fractionated and isolated by preparative TLC. The cytotoxic activity and IC50 of this fractions were analyzed. The results showed that only 2 from 9 crude extracts that able to calculate its IC50 because those two extracts have concentration dependent pattern of inhibition concentration. Chloroform extract have the lowest IC50 value (369,837 μg/mL). Then, this extract fractionated by eluen n-hexane : ethyl acetate 4:1. Four fractions were collected. The lowest IC50 value is fraction IV (144,41 μg/mL). Based on the results it could be concluded that S. plicata leaves have moderate potency to develop as anticancet agents, especially on breast cancer. Keywords: S. plicata, T47D cells, cytotoxic, MTT assay, preparative TLC. 


Author(s):  
Nur Ismiyati ◽  
Dyaningtyas Dewi Pamungkas Putri ◽  
Siska Andrina Kusumastuti ◽  
Rifki Febriansyah

Eugenia uniflora Lam. is one of herbal products developed for anticancer. The aim of the present study was to identify the antiproliferative effect of ethanolic extract of Eugenia Uniflora Lam. leaves (EEU) on breast cancer cell line T47D. This Research was initiated by extracting the active contents of Eugenia uniflora Lam. leaves by maceration with ethanol 96%. The extract was then analyzed by thin layer chromatography (TLC). Cytotoxic assay of EEU was carried out by using MTT assay. Apoptosis phenomenon was observed with double staining using acridine orange-ethidium bromide. EEU showed cytotoxic effect on T47D cells with IC50 value of 65 µg/ml. Moreover, EEU 50µg/ml and 100µg/ml induced apoptosis. TLC examination showed that EEU used in this study contain phenolic, flavonoid, and saponin compounds which were suggested to be responsible for antiproliferative effect. Further molecular mechanism underlying EEU antiproliferative effect needs to be done.Keywords: Eugenia uniflora Lam., T47D cells, antiproliferative, apoptosis


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