Cytotoxic Assay of Semipolar Fraction Of Ethanolic Extract From Sugar Apple (Annona Squamosa L.) Stem Bark on T47D Cells

Previous research has shown that some compounds in leaves and seeds of sugar apple have a cytotoxic activity. The aim of this research was to determine the cytotoxicity of semipolar fraction of ethanolic extract from sugar apple stem bark (Annona squamosa L.) on T47D cancer cells. The semipolar fraction of ethanolic extract from sugar apple stem bark was collected by fractionation using Vacuum Liquid Chromatography (VLC) with hexane:ethyl acetic (9:1, 8:2, 7:3, and 6:4) as mobile phase. Cytotoxicity from the fractions of five different concentration namely; 25, 50, 100, 150, and 250, µg/mL was measured by MTT assay. The potency of the cytotoxicity was defined by the ability of the fraction to inhibit the growth of T47D cells indicated by the value of IC50. Qualitative analysis of contained compounds in the fraction was done by Thin Layer Chromatography (TLC) method using silica gel F 254 as a stationary phase and hexane:ethyl acetic (7:3) as a mobile phase. UV 254 and 366 nm lamp also Dragendorff, citroboric, and FeCl3 spray reagents were used to visualize the spots of the secondary metabolites. The result proved that the semipolar fraction of ethanolic extract from sugar apple stem bark showed potential cytotoxicity on T47D cancer cells with IC50value of 70,77 µg/mL. Qualitative analysis showed that the fraction contained flavonoids and alkaloids which is presumably responsible for its cytotoxic activity.

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Synergistic Combination of Ciplukan (Physalis angulata) Herbs Ethanolic Extract and Doxorubicin on T47D Breast Cancer Cells

Indonesian Journal of Cancer Chemoprevention ◽

10.14499/indonesianjcanchemoprev1iss1pp26-31 ◽

2010 ◽

Vol 1 (1) ◽

pp. 26

Author(s):

Inna Armandari ◽

Kartika Dyah Palupi ◽

Sofa Farida ◽

Adam Hermawan ◽

Ratna Asmah Susidarti ◽

...

Keyword(s):

Breast Cancer ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Cardiac Toxicity ◽

Chemotherapeutic Agent ◽

Ethanolic Extract ◽

High Dose ◽

T47d Cells ◽

Ic50 Value

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

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Secang (Caesalpinia sappan L.) Heartwood Ethanolic Extract Shows Activity as Doxorubicin Co-chemotherapeutic Agent by Apoptotis Induction on T47D Breast Cancer Cells

Indonesian Journal of Cancer Chemoprevention ◽

10.14499/indonesianjcanchemoprev3iss2pp376-383 ◽

2012 ◽

Vol 3 (2) ◽

pp. 376 ◽

Cited By ~ 1

Author(s):

Ika Nurzijah ◽

Dyaningtyas Dewi Pamungkas Putri ◽

Erlina Rivanti ◽

Edy Meiyanto

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Chemotherapeutic Agent ◽

Annexin V ◽

Ethanolic Extract ◽

Multi Drug Resistance ◽

Caesalpinia Sappan ◽

T47d Cells ◽

Induced Apoptosis

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

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CYTOTOXIC EFFECT OF AQUADEST, ETHANOLIC, AND CHLOROFORM EXTRACTS OF Spathoglottis plicata Blume ON BREAST CANCER CELLS LINE (T47D CELLS)

KnE Life Sciences ◽

10.18502/kls.v2i1.118 ◽

2015 ◽

Vol 2 (1) ◽

pp. 64

Author(s):

Mukhlish Jamal Musa Holle ◽

Hestri Dyah Puspitasari ◽

Andaru Satryo ◽

Wahyu Dewi Astuti Ningrum ◽

Digdo Sudigyo ◽

...

Keyword(s):

Breast Cancer ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Mtt Assay ◽

Breast Cancer Cells ◽

Chloroform Extract ◽

Probit Analysis ◽

Antioxidant Compounds ◽

T47d Cells ◽

Ic50 Value

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.

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The Anticancer Activity of Srikaya Leaves Fraction (Annona squamosa L.): An In Vitro Study

BioScientia Medicina ◽

10.32539/bsm.v3i4.102 ◽

2019 ◽

Vol 3 (4) ◽

pp. 39-44

Author(s):

Makbruri Amin ◽

Irsan Saleh ◽

Rachmat Hidayat

Keyword(s):

Cytotoxic Activity ◽

Anticancer Activity ◽

Anticancer Drugs ◽

T Test ◽

Therapeutic Effects ◽

Annona Squamosa ◽

Methanol Fraction ◽

T47d Cells ◽

Average Percentage ◽

Ic50 Value

Abstract Introduction: Anticancer drugs are aimed primarily at inhibiting the growth and proliferation of cancer cells. Srikaya leaves (Annona squamosa L.) had been proven to possess various therapeutic effects and potential to be developed as anticancer drugs due to its cytotoxic activity. Aim of study: This study aimed to assess the anticancer activity of srikaya leaves (Annona squamosa L.) fraction. Methods: Methanol fraction of srikaya leaves were obtained at concentrations of 500; 250; 125; 62.5; 31.25 Âµg/ml. Srikaya methanol fraction and cisplatin as control were given to a plate that was sealed with T47D cells for MTT assay. Identification of compounds in the methanol fraction of srikaya leaves was performed with thin layer chromatography (TLC). Data were collected in the form of absorbance value and half-maximal inhibitory concentration (IC50) value was determined â€‹â€‹by linear regression. Data analysis was carried out with paired T test, unpaired T test, and ANOVA. Results: Average percentage of T47D cells viability increased with the decrease in the concentration of srikaya methanol fraction. Obtained IC50 value was 174.25 Âµg/ml which was quite active and potential to be developed as an anticancer drug. Methanol fraction of srikaya leaves contained secondary terpenoid metabolites, steroids, phenols, flavonoids, alkaloids and tannins. Flavonoid was the dominant metabolites in phytochemical tests and believed to play a major role in cytotoxic activity of srikaya leaves. Conclusion: Methanol fraction of srikaya leaves possessed the cytotoxic effect on T47D cancer cell line through the role of flavonoid metabolites. Keywords: srikaya, Annona squamosa, anticancer, T47D cells

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Efek Sitotoksik Ekstrak dan Fraksi Umbi Paku Atai Merah (Angiopteris ferox Copel) Terhadap Sel Kanker Payudara T47D

Jurnal Farmasi Galenika (Galenika Journal of Pharmacy) ◽

10.22487/j24428744.2020.v6.i2.15255 ◽

2020 ◽

Vol 6 (2) ◽

Author(s):

Andi Nur Aisyah ◽

Syamsu Nur ◽

Endang Lukitaningsih ◽

Rumiyati Rumiyati ◽

Asril Burhan ◽

...

Keyword(s):

Breast Cancer ◽

Ethyl Acetate ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Supportive Therapy ◽

Ethanolic Extract ◽

Ethanol Extract ◽

Ethyl Acetate Fraction ◽

Screening Tests

The use of natural products has been widely used as a resource of new bioactive chemical compounds. One of them is the Paku Atai Merah (Angiopteris ferox Copel) tuber which has long been used empirically by the Dayak tribe of East Kalimantan as an anti-cancer. The purpose of this study was to determine the anticancer cytotoxic activity of the extract and fractions of Paku Atai Merah tuber against T47D breast cancer cells in vitro. Extract of Paku Atai Merah tubers was obtained by maceration method using ethanol solvent until obtained the ethanolic extract then fractionated using various solvents to obtain n-hexane, ethyl acetate, and aqueous-ethanol fractions. The cytotoxic effect was carried out based on the MTT assay. Phytochemical screening tests showed positive results for the presence of flavonoid, phenolic, tannin, saponin and steroid compounds. The results of the cytotoxic activity study showed that the ethyl acetate fraction had moderate cytotoxic activity in T47D cancer cells with an IC50 value of 84.8 µg/ml. Ethanol extract (513.06 µg/ml) and n-Hexane frsction (881.97 µg/ ml) were also included in the weak category. This study indicates that ethyl acetate fraction can be developed as a supportive therapy for breast cancer treatment.

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Identifikasi Isolat Ekstrak Etanol Kulit Batang Rhizophora apiculata Blume dan Rhizophora mucronata Lam Serta Sitotoksisitasnya Terhadap Sel MCF-7 dan T47D

Pharmacon Jurnal Farmasi Indonesia ◽

10.23917/pharmacon.v18i01.12252 ◽

2021 ◽

Vol 18 (1) ◽

pp. 9-22

Author(s):

Joko Kristianto ◽

Haryoto Haryoto ◽

Peni Indrayudha

Keyword(s):

Ethyl Acetate ◽

Cytotoxic Activity ◽

Silica Gel ◽

Methanol Extract ◽

Ethanol Extract ◽

Ethyl Acetate Fraction ◽

Stem Bark ◽

T47d Cells ◽

Static Phase ◽

Mcf 7

Mangrove plants as traditional medicine have long been used by the society for the therapy of anticancer diseases. So far, the potency of mangrove plants as anticancer has not been studied intensively. The methanol extract of R. mucronata stem bark has cytotoxic activity on myeloma cells. This study aims to determine the isolates of the ethyl acetate fraction of the methanol extract of the stem bark of R. mucronata and R. apiculata, and to examine the potency of cytotoxic activity against MCF-7 and T47D cells. The powder of the stem bark of R. mucronata and R. apiculata were extracted with methanol by maceration, and then the methanol extract was fractionated consecutively using n-hexane, ethyl acetate, and methanol. Ethyl acetate fraction was then purified by column chromatography with a diameter of 2 cm and a height of 30 cm with a static phase in the form of silica gel 60 (0.063-0.2 mm) Merck 7734 and eluent n-hexane:ethyl acetate (6: 4, 5: 5, 6: 4, 7: 3, 8: 2, 9: 1) and n-hexane: methanol (9,5: 0,5) 10 ml each. The eluates that produce the same number and pattern of stains were combined into one isolate. Then the eluate was evaporated at room temperature. Then the purification results of the extract R. mucronata were isolated using preparative TLC with 0.25 mm GF254 silica gel as static phase and n-hexane mobile phase: ethyl acetate (6: 4), while the apiculata extract with n-hexane mobile phase: ethyl acetate (5: 5). The results of the stain separation of the compound were scraped off and separated from the static phase using a solvent. The purity of isolates compound was examined using TLC in the static phase of silica gel GF254 0.25 mm. The qualitative test results of the ethyl acetate fraction of R. apiculata stem bark with an NMR spectrophotometer showed a composition of cis-3- (3,5-dihydroxyphenyl) acrylic acid, while the ethyl acetate fraction of R. mucronata stem bark contained n-hexan-3-ol compounds. The cytotoxicity test of ethanol extract, n-hexane fraction, ethyl acetate fraction and ethanol extract was carried out on MCF-7 and T47D cells using the MTT method. The results proved that the ethanol extract, n-hexane fraction, ethyl acetate fraction and ethanol fraction from the ethanol extract of R. mucronata and R. apiculata stem bark, based on cytotoxic activity, were inactive against MCF-7 and T47D cells.

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New curcumin analog, CCA-1.1, synergistically improves the antiproliferative effect of doxorubicin against T47D breast cancer cells

INDONESIAN JOURNAL OF PHARMACY ◽

10.22146/ijp.681 ◽

2020 ◽

pp. 244-256

Author(s):

Febri Wulandari ◽

Muthi' Ikawati ◽

Dhania Novitasari ◽

Mitsunori Kirihata ◽

Jun-ya Kato ◽

...

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Cytotoxic Activity ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Cell Cycle Progression ◽

Synergistic Effects ◽

Annexin V ◽

Curcumin Analog ◽

T47d Cells

An improved compound of pentagamavunone-1 (PGV-1), chemoprevention-curcumin analog 1.1 (CCA-1.1), has been synthesized and proven to have antiproliferative effects against breast cancer cells. This study is designed to investigate the potency of CCA-1.1 alone and in combination with doxorubicin (Dox) on T47D cells in comparison with that of PGV-1. We used the MTT assay to assess cytotoxic activity. Propidium iodide (PI), annexin-V–PI, and DCFDA staining were respectively used to determine cell cycle profiles, apoptosis, and intracellular reactive oxygen species (ROS) levels. Senescent cells were identified using the SA-b-galactosidase assay. Our results revealed that CCA-1.1 possesses cytotoxic effects similar to those of PGV-1 on T47D cells. Synergistic effects during co-treatment with Dox were also observed. CCA-1.1 arrested cell cycle progression at the G2/M phase and limited sub-G1 accumulation, which is correlated with apoptosis. CCA-1.1 alone and in combination with Dox increased senescence and intracellular ROS to a similar level to those achieved by PGV-1. CCA-1.1 alone and in combination with Dox enhanced cytotoxic activity toward T47 cells compared to PGV-1. Thus, this curcumin analog may be a potential chemotherapeutic/co-chemotherapeutic candidate for estrogen receptor-positive (ER+) breast cancer therapy.

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