Synthesis and Cytotoxic Activity of Methoxylated Chalcones in Breast Cancer MCF-7 and Prostate Cancer DU-145 Cell Lines

BackgroundLoss of BIN1 tumor suppressor expression is abundant in human cancer and its frequency exceeds that of genetic alterations, suggesting the role of epigenetic regulators (DNA methylation). BIN1 re-expression in the DU145 prostate cancer cell line after 5-aza-2'-deoxycytidine treatment was recently reported but no methylation of the BIN1 promoter CpG island was found in DU145.MethodsMethylation-sensitive arbitrarily-primed PCR was used to detect genomic loci abnormally methylated in breast cancer. BIN1 CpG island fragment was identified among the differentially methylated loci as a result of direct sequencing of the methylation-sensitive arbitrarily-primed PCR product and subsequent BLAST alliance. BIN1 CpG island cancer related methylation in breast and prostate cancers was confirmed by bisulphite sequencing and its methylation frequency was evaluated by methylation sensitive PCR. Loss of heterozygosity analysis of the BIN1 region was performed with two introgenic and one closely adjacent extragenic microsatellite markers.BIN1 expression was evaluated by real-time RT-PCR.ResultsWe have identified a 3'-part of BIN1 promoter CpG island among the genomic loci abnormally methylated in breast cancer. The fragment proved to be methylated in 18/99 (18%) and 4/46 (9%) breast and prostate tumors, correspondingly, as well as in MCF7 and T47D breast cancer cell lines, but was never methylated in normal tissues and lymphocytes as well as in DU145 and LNCaP prostate cancer cell lines. The 5'-part of the CpG island revealed no methylation in all samples tested. BIN1 expression losses were detected in MCF7 and T47D cells and were characteristic of primary breast tumors (10/13; 77%), while loss of heterozygosity was a rare event in tissue samples (2/22 informative cases; 9%) and was ruled out for MCF7.ConclusionBIN1 promoter CpG island is composed of two parts differing drastically in the methylation patterns in cancer. This appears to be a common feature of cancer related genes and demands further functional significance exploration. Although we have found no evidence of the functional role of such a non-core methylation in BIN1 expression regulation, our data do not altogether rule this possibility out.

Date seeds are rich in many dietary elements, antioxidants, and anti-inflammatory and anticancer phytochemical compounds. The present study aimed to evaluate the anticancer potential of Iraqi Date Palma dactylifera L. seed extracts on breast cancer MCF7 and prostate cancer PC3 cell lines. The seeds were extracted with 70% ethanol and explored for the presence of many anticancer compounds by High-performance liquid chromatography (HPLC) analysis . The cytotoxicity of date seed extracts on cell lines was studied using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. The cytotoxic assay of multi-parameters was carried out to detect the extract activity on valid cell count, total nuclear intensity, cell membrane permeability, mitochondrial membrane potential and cytochrome C release by using High-Content Screening (HCS) assay. The results of HPLC confirmed the presence of caffeic acid, sinapic acid and gallic acid. The results of MTT assay showed significantly reduced cell viability of MCF7, PC3 and control HdFn cell lines with IC50 of 269,325 and 1499 μg/ml, respectively. Analysis of HCS findings indicated significant changes in all tested parameters at concentrations of 100 μg/ml. Ethanolic extract of seeds was rich in many antioxidant compounds and the extract appeared to have strong cytotoxic activity against both MCF7 and PC3 cancer cell lines. The cells of MCF7 were more sensitive to extract than PC3. The viability of normal HDFn cells was not affected by the extract. The study showed the importance of date seeds as a very effective antioxidant and can contribute to reducing the risk of breast and prostate cancer.

Aim: Ancient Indian medicine has long employed the xerophytic plant Aloe vera (Family: Liliaceae) to treat several diseases and disorders, including diabetes and cancer, despite the limited scientific validation of these claims. This study aimed to identify and characterize secondary metabolites from the ethanol extract of A. vera leaves and assess their anti-cancer effects. In-vitro cytotoxicity was assessed against human OAW42 (ovarian) and MCF-7 (breast) cancer cell lines, while the in-vivo antitumor efficacy was evaluated using the Ehrlich ascites carcinoma (EAC) tumour model in Wistar rat model. Background: Among two prepared fractions of Aloe vera extraction such as ethyl acetate soluble fraction (EAF) and the chloroform fraction (CF), EAF executed higher potency compared to others and was further utilized to isolate bioactive compounds, Barbaloin (1) and Gallic acid (2), which exhibited significant anti-cancer activity in both in-vitro and in-vivo assays. We further examined the antitumor activity of the ethyl acetate extract and the isolated compounds effectively modulated tumour-associated parameters and restored various hematological indices in EAC tumour-bearing rat. Objectives: The basic objective of this research work is to describe the significant molecular mechanism of barbaloin and gallic acid against the cancer activity against MCF-7 and OAW42 cancer cell lines in-vitro and in a Wistar rat model in-vivo. Methods: Barbaloin and gallic acid demonstrated dose-dependent growth inhibition along with the significant IC50 values for barbaloin and gallic acid against MCF-7 and OAW42 cell lines. Flow cytometry revealed that both compounds induced apoptosis and G2/M cell cycle arrest. Western blot analysis exhibited modulation of apoptosis and cell cycle regulatory proteins. In-vivo studies in Wistar rats with Ehrlich ascites carcinoma demonstrated that barbaloin and gallic acid treatment decreased the significant parameters of cancers compared to controls. Results: Comparing with barbaloin and gallic acid with their individual effect to the combination of those, the combination of these compounds exhibits promising anti-cancer activities both in the study of in-vitro and in-vivo. Tumor volume is quantified based on the volume of ascitic fluid, the number of viable tumor cells present in that fluid which can be surgically removed from the animal body. Conclusion In conclusion, this study demonstrates that barbaloin and gallic acid individually possess significant anti-cancer activities individually and in combination against MCF-7 breast cancer and OAW42 ovarian cancer cell lines in-vitro and in a Wistar rat model of Ehrlich ascites carcinoma in-vivo. The combination of barbaloin and gallic acid significantly decrease the viable cell count and increase the non-viable cell count.

The combination therapy of breast cancer has preferred for the patients to minimize possible side effects, drug resistance, recurrence and toxic effects. In this study, we aim to investigate the cytotoxic and antitumor activities the tamoxifen and doxorubicin combination in breast cancer cell lines, MCF-7 and BT-474. Tamoxifen (Tam) and doxorubicin (Dox) and their combination with different concentrations (0.625–20 μM Tam; 0.0625–2 μM Dox and 5 μM Tam+ 0.5/1.0/1.5 μM Dox combination were applied to MCF-7 and BT-474 cells for 48 hours. Afterthat, their cytotoxic activities were analyzed with MTT assay. Bcl-2, Mapt and Mrp1 are genes that induce cell proliferation, inhibit apoptosis and play role in drug resistance in cancer cells. To evaluate the antitumor activities of these genes in combination treatment, mRNA levels were analyzed by quantitative PCR. According to the MTT assay, it was determined that IC50 values as 17.26 μM and 16.65 μM for tamoxifen on MCF-7 and BT-474 breast cancer cell lines. IC50 values of doxorubicin in MCF-7 and BT-474 cells were 1.65 μM and 1.57 μM, respectively. It was found that the application of the combination drugs (15 μM tamoxifen and 1.5 μM doxorubicin) in MCF-7 and BT-474 cells have the lowest combination index values as 1.09 and 1.26, respectively. Therefore, the combination of 15 μM tamoxifen and 1.5 μM doxorubicin was selected and applied to both breast cancer cell lines for gene expression analysis. It was found that while Mrp1 and Mapt genes expressions were significantly upregulated, Bcl-2 gene expression was downregulated in MCF-7 cells. However, Bcl-2, Mrp1 and Mapt genes expressions in BT-474 cells were not significantly regulated. Altogether, these findings suggest that the combination of these two drugs may have a potential antagonistic interaction according combination index values.

Cryptocarya pulchrinervia is an Indonesian indigenous plant that grows in Sumatra, Kalimantan and Papua. One of the new compounds extracted from this plant was cryptobrachytone C, which was known to be cytotoxic against cancer cells of Murine leukemia P388 with IC50 10.52 μM. In this study, the cytotoxicity and anticancer properties of cryptobrachytone C on proliferation, apoptosis, migration and clone formation of MCF-7 and T47D breast cancer cell lines were examined, which had not previously been done before. The cytotoxicity of the compound was measured using an MTT (3- (4,5-dimethylthiazol-2- yl) -2,5-di-phenyl-tetrazolium bromide) assay. The cell proliferation was measured using a BrdU assay, and the cell apoptosis was measured using annexin-V FITC, while the cell migration was measured using a transwell filter. The cytotoxic test result demonstrated that cryptobrachytone C was cytotoxic against MCF-7 cells with IC50 12.94 ± 0.32 μM but not against T47D cells with IC50 65.33 ± 2.33 μM nor against normal MRC-5 cells with IC50 122.57 ± 19.84 μM. The cell proliferation assay showed that cryptobrachytone C at IC50 concentration had antiproliferative properties against MCF-7 cancer cell lines (p < 0.05) but did not significantly reduce T47D cell proliferation (p < 0.07). Although the results of the cell apoptosis test showed that cryptobrachytone C could induce the apoptosis of the MCF-7 and T47D cells, it was insignificant (p > 0.05). The cell migration test showed that cryptobrachytone at IC50 concentrations could inhibit the migration of the MCF-7 and T47D cells. The clonogenic test showed that cryptobrachytone C at IC50 concentration can induce the inhibition of the formation of MCF-7 and T47D cell colonies. The cryptobrachytone C anti-cancer character was more signi icant on the MCF-7 cell line compared to the T47D. This study showed that cryptobrachytone C was cytotoxic and had potential as an anti-cancer compound against MCF-7 and T47D breast cancer cell lines.

Differentially expressed proteins among cancer cell lines fit each cell line as a model for gaining knowledge of heterogeneity in cancer. The aim of our study was to identify differentially expressed proteins between MCF7 (ER+, low HER2 expression) and SKBR3 (ER-, high HER2 expression) cell lines by a proteomic approach. Moreover, a number of proteins in MCF7 cell line were randomly selected and identified. MCF7 and SKBR3 cell lysates were subjected to two dimensional gel electrophoresis and spots of interest were identified by MALDI-TOF/TOF mass spectrometry. Upregulated proteins (≥2 fold and p value &amp;lt;0.05) in MCF7 cells were cellular retinoic acid binding-protein 2, Hsp27, nucleophosmin, electron transfer flavoprotein ≤ subunit, and profilin, and in SKBR3 cells were Rho GDP dissociation inhibitor-α (RhoGDI-α), voltage-dependent anion channel 2, aldehyde dehydrogeanase-2 (ALDH2), LDH- A, LDH-B, pyrophosphates 1, GAPDH, cathepsin D preprotein, and apolipoprotein A-I binding. Most of the identified proteins have been a candidate marker for cancer aggressiveness or drug resistance, but their differential expressions between SKBr3 and MCF7 cells were not known. Apo-lipoprotein binding-protein has not been described in cancer so far. Differential expression of RhoGDI-α was further validated by using western blotting with specific antibody. Further studies are required to clarify the importance of differential expressions of the identified proteins in SKBr3 and MCF7 breast cancer cell line models. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4785. doi:1538-7445.AM2012-4785

Sophora pachycarpa Schrenk ex C.A.Mey. is an annual plant belonging to the family Fabaceae. The cytotoxic activities of methanol-dichloromethane extracts (1:1) of different parts of S. pachycarpa were investigated on DU145 (prostate cancer cell line) and MCF-7 (breast cancer cell line) cell lines. The root extract of S. pachycarpa was the only extract that showed significant cytotoxic activity with IC50 values of 39.88 and 16.49 μg/mL on DU145 and MCF-7 cell lines, respectively. The root extract was then subjected to RP-HPLC for further fractionations. Among the isolated fractions from root extract, only one of them had remarkable cytotoxic effects with IC50 value of 26.43 on MCF-7 and 7.54 μg/mL on DU145 cell lines. Further purification led to isolation of a compound with IC50 values of 5.44 and 2.44 μg/mL on MCF-7 and DU145 cell lines, respectively. Based on 1H NMR and 13C NMR spectra, together with LC-MS, the structure of the purified compound was assigned as the flavonostilbene alopecurone A.

Background: Breast cancer is the most common cause of cancer-related death in women worldwide. Therefore, there is an urget need to identify and develop therapeutic strategies against this deadly disease. This study is the first to investigate the effects of Hemolymph Serum of Potamon persicum Crab (HSPPC) on MCF-7 and MDA-231 breast cancer cell lines. Materials and Methods: LDH and MTT assays were performed on MCF-7 and MDA-231 breast cancer cell lines as well as human umbilical vein endothelial cells (HUVEC) to determine the cytotoxic and antiproliferative activity of the HSPPC at different concentrations. Further, the apoptosis inducing action of the hemolymph serum was determined by TUNEL (terminal deoxynucleotidyl transferasemediated dUTP nick end labeling) and cell death assay. Results: The IC50 values of HSPPC for MCF-7 and MDA-231 cell lines were 960±0.369 and 850±1.422 μg/mL, respectively. The growth of both MCF-7 and MDA-231 cell lines were significantly (P&lt;0.001) inhibited by HSPPC as compared with untreated controls at 48 hours. The results showed that HSPPC had no cytotoxic effects but significantly inhibited cell growth in a dose and time dependent manner. In addition, DNA fragmentation analysis (TUNEL) and cell death assay indicated induction of apoptosis by HSPPC in MCF-7 and MDA-231 cell lines. Conclusion: The results suggest that HSPPC contains bioactive compound(s) with potentials for the treatment of breast cancer.

Development of certain aminoquinazoline scaffolds as potential multitarget anticancer agents with apoptotic and anti-proliferative effects: Design, synthesis and biological evaluation

1,3,5-triazine derivatives as potential anticancer agents against lung and breast cancer cell lines: Synthesis, biological evaluation, and structure-based drug design studies

Recombinant human tumor necrosis factor-related apoptosis-inducing ligand (rhTRAIL), the optimized form of the endogenous death ligand TRAIL, shows therapeutic potential for cancer due to its ability to induce apoptosis in cancer cells independent of p53, while exhibiting minimal toxicity to normal cells. Despite this, a majority of breast cancers display resistance to rhTRAIL treatment due to up-regulation of pro-apoptotic proteins, down-regulation of anti-apoptotic proteins, and/or up-regulation of death receptors (DR) 4 and 5. To overcome rhTRAIL resistance, natural compounds have been investigated as sensitizing agents. This study considered the application of the naturally occurring flavonol Quercetin (Q). Q has been shown to have the ability to up-regulate DR5 and down-regulate anti-apoptotic proteins in cancer cells, and thereby, making Q a favorable choice to be employed as a sensitizing agent. The intention of this study was to ascertain the capacity of Q to sensitize rhTRAIL-resistant triple negative breast cancer BT-20 cells and hormone-dependent breast cancer MCF-7 cells to rhTRAIL-induced apoptosis and elucidate the underlying mechanism for Q’s sensitization. Q demonstrated the ability to intensify rhTRAIL’s pro-apoptotic effects in the breast cancer BT-20 and MCF-7 cell lines as detected through Annexin V/PI assays followed by FACS analysis. In comparison to single agent treatments, the cotreatment of Q and rhTRAIL enhanced the induction of the extrinsic pathway of apoptosis as marked by PARP cleavage (a hallmark of apoptosis), activation of caspase 8, and activation of the executioner caspases 3 and 7. The mechanism for Q’s augmentation in breast cancer was determined to be through the down-regulation of c-FLIPL (caspase 8 inhibitor) in a dose-dependent manner. Furthermore, Q promoted the ubiquitination of c-FLIPL facilitating the proteasome-mediated degradation of c-FLIPL in breast cancer. An additional mechanism for Q’s sensitization was displayed in breast cancer BT-20 cells. Q up-regulated DR5 membrane and protein expression in breast cancer BT-20 cells in a dose-dependent manner. RT-PCR analysis revealed that Q’s influence on DR5 expression occurred at the transcriptional level in those breast cancer cells. Thus, these data suggest that the cotreatment of Q and rhTRAIL possesses the therapeutic potential to be an effective anti-breast carcinoma regimen. Citation Format: Jasmine M. Manouchehri, Michael Kalafatis. TRAIL-induced apoptosis in TRAIL-resistant breast carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2117. doi:10.1158/1538-7445.AM2017-2117

Aims: The sour cherry pit oil (SCPO) displays the potent anti-inflammatory, and antioxidant activities. In the present study, we have produced the SCPO nanoemulsion (SCPO-NE) to evaluate their anticancer impacts on breast cancer comparing with its un-processed oil.Methods: We employed an ultrasonication method to formulate the stable SCPO-NE. Their size, stability, and morphology were measured. Then, their cytotoxic impacts and apoptotic activity were checked on MCF7 breast cancer cells and compared with the normal Human foreskin fibroblasts (HFF). Finally, their anti-tumour effect was studied on murine breast cancer model (inoculated with TUBO cancer cells).Results: The results indicated the 36.5 nm stable SCPO-NE significantly decreased the MCF7 cells viability comparing with normal HFF cells, and reduced the tumour size in the murine model.Conclusion: We suggest that SCPO-NEs are able to efficiently inhibit breast cancer progression in both MCF7 cells and murine breast cancer model through apoptotic death induction.

Despite the fact that the ovaries stop to produce oestrogens at menopause, high levels can be detected locally in breast tumours. There are several enzymes responsible for the local production of sex steroids such as aromatase, and the 17 beta hyroxysteroid dehydrogenase (17βHSD) family has also proven to have a significant importance in breast cancer. 17βHSD1 converts estrone to estradiol, 17βHSD2 estradiol to estrone, 17βHSD5 androstendione to testosterone and in the next step aromatase is responsible for formation of testosterone to estradiol, the function of 17βHSD14 is still not clear. The regulation of 17βHSDs needs further investigation. In several studies combined hormone replacement therapy (estrogens and progestins) has shown an increased risk to develop invasive breast cancer, compared to women treated with only estrogens. It is plausible that progestins influence the expression of members in the 17βHSD-family. Previous studies investigating the role of progestins on steroid conversion have focused on measuring steroidal levels and have not investigated changes in expression levels of enzymes responsible for this conversion. The aim of this study was to treat breast cancer cell lines with progesterone and progestins to detect possible influence on 17βHSD1, 17βHSD2, 17βHSD5 and 17βHSD14 mRNA expression. We further investigated conversion of androstendione in cultured media. The breast cancer cell lines T47D, MCF7 and ZR75-1, choosen for their different endogenous expression of progesterone receptor (PgR) were exposed for progesterone, medroxyprogesterone acetate (MPA) or levenogestrel for 48 and 72 hours at 10-7 and 10-9 M. The mRNA expression of 17βHSD1, 17βHSD2, 17βHSD5 and 17βHSD14 were measured by real time PCR using assay on demand and ACTH as endogenous control. Conversion of H3 labelled androstendione was investigated using HPLC coupled radio detector. We found that mRNA expression of 17βHSD1 increased significantly after 48 h in MPA treated T47D (10-9 M p=0.03) and MCF7 cells (10-7 p=0.001; 10-9 p=0.004). Further, mRNA expression of 17βHSD5 increased after 48 h in MPA treated T47D (10-7 M p=0.0006; 10-9 M p=0.04) and MCF7 cells (10−9 M p=0.005). No significant differences were found in cells treatment with levonogestrel or progesterone. We did neither find any differences in the mRNA expression levels of 17βHSD2 nor 17βHSD14. In the ZR75-1 cell line no significant changes were detected. We have preliminary results indicating that androstendione is converted in T47D cells treated with MPA. In conclusion, we show that reductive members of the 17βHSD-family increase (17βHSD1 and 17βHSD5) in breast cancer cells with high PgR expression levels. Our results may indicate that 17βHSD5 can be involved in the high risk for breast cancer assessed with hormone replacement treatment using combined estrogen and progestin in post menopausal women. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2286. doi:10.1158/1538-7445.AM2011-2286

BackgroundNotch signalling is normally involved in lateral inhibition during embryogenesis and is found to be de regulated in many breast cancers. Gamma-secretase is an enzyme complex formed by constitution of Nicastrin, APH1, Presenilin and Pen2 and is involved in the cleavage of notch proteins. We investigated the effects of γ-secretase inhibition and its mechanism of action in breast cancer cell lines.Materials and MethodsGrowth assay was performed using Gammasecretase inhibitor-1(Z-Leu-Leu-Nle-CHO) in MCF-7, MDA-MB-231, T47D and ZR-75 breast cancer cell lines and 226 U19 and MCF10A non tumourigenic epithelial cell lines at concentrations between10nM and 60uM. Expression of Bcl-2, Bcl-XL, P-Bad and XIAP were measured by Western blot and cell cycle analysis was done by FACS in MCF-7 and MDA-MB-231 cells treated with GSI-1 at 0.75μM, 2μM and 5μM for 48 hours. Caspase 3 and 7 activity was measured in MCF-7 and MDA-MB-231 cells treated with GSI-1 at 0.75μM, 2μM and 5μM for 24 and 48 hours using Promega assay kit. MCF-7 cells were also treated with 0.75μM GSI-1 and DMSO in five replicates for 48 hours and gene array was performed using 44K Agilent array. Fold changes of AKR1B10 and Osteopontin genes were confirmed by Real time PCR at 0.75μM of GSI-1 in MCF-7 and they were also measured at concentrations of 2μM and 5μM in MCF-7 and MDA MB 231 cells using Taqman.ResultsThe 4 cancer cell lines MCF-7, MDA-MB231, ZR-75 and T47D showed growth inhibition in a dose dependant manner and were killed at 5uM. However this concentration did not affect the non tumourigenic cell lines. Expression of Bcl-2, Bcl-XL, inhibitor of apoptosis (XIAP) and P-BAD went down with escalating doses of GSI-1 at 48 hours in both MCF-7 and MDA MB 231 cells. Cell cycle analysis confirmed increase in the percentage of cells in G2M arrest leading to apoptosis at 48 hours in the same cell lines. However we observed increase in Caspase 3 and 7 activity only in MDA-MB-231 not in MCF-7 cells. Gene array in MCF-7 cells treated with GSI1 showed 15 and 4 fold changes in the expression of AKR1B10 and Osteopontin. There was also dose dependant increase in the expression these two genes with GSI-1 in MCF-7 but no such change was observed in MDA MB 231 cells.DiscussionThis is the first time we are reporting the effect of GSI-1 and its possible mechanism of action in breast cancer cell lines. Apoptosis appears to be the main mechanism of cell death as evidenced by our Western blots and FACS analysis in MCF-7 and MDA MB 231 cell lines. However, MCF-7 did not show any Caspase 3/7 activity implying other mechanisms may be involved in the induction of apoptosis. Production of reactive oxygen species (ROS) triggers the damage to mitochondrial membrane and release of cytochrome c, Apoptosis inducing Factor and Endonuclease G resulting in the initiation of Caspase independent apoptosis. AKR1B10 and Osteopontin were shown to be highly expressed in conditions resulting in increased production of ROS. Thus inhibition of γ-secretase enzyme results in breast cancer cell death in a dose which was non lethal to non tumourigenic cell lines. Both Caspase dependant and independent apoptosis appear to be involved in the mechanism of cell death. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3129.

Breast cancer is one of the cancers with the highest incidence and mortality rate in the world. Curcuma caesia (Black turmeric) is known to have neuroprotective, anthelmintic, sunscreen activity, anti-diabetic and also anti-cancer activity. This study aimed to determine the phytochemical content and possible cytotoxic activity of C. caesia water extract (EWCC) and C. caesia ethanol extract (EECC) on breast cancer cell lines. Samples were extracted using two extraction methods, maceration technique using 96% ethanol as solvent and double boiling using water as a solvent. The phytochemical content, total phenolic content (TPC), and total flavonoid content (TFC) of each extract were determined using spectrophotometer method. Cytotoxic activity was measured using MTT assay on MCF-7 and 4T1 breast cancer cell lines (24 hrs and 48 hrs). Results showed that EWCC and EECC contained flavonoids, alkaloids, and polyphenols. The values of TPC and TFC were 106.08 mg GAE/100 g and 4.32 mg QUE/g for EECC, respectively, and 73.29 mg GAE/100 g and 3.40 mg QUE/g for EECC, respectively. Meanwhile, the cytotoxic activity test found that EWCC did not provide cytotoxic activity on MCF-7 and 4T1 cells. In the MCF-7 cell line, EECC exhibited IC50 values of 212.90 µg/mL at 24 hrs and 311.91 µg/mL at 48 hrs. Similarly, in the 4T1 cell line, EECC exhibited IC50 values of 161.06 µg/mL at 24 hrs and 150.89 µg/mL at 48 hrs. From these results, it can be concluded that EECC has cytotoxic activity in MCF-7 and 4T1 breast cancer cell models, so this extract can be studied more deeply to determine its pathway of action.