• 한국식품영양과학회지
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• 제36권11호
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• pp.1385-1390
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• 2007

당귀로부터 항암활성을 화합물을 silicagel column chromatography를 이용하여 분리 및 정제한 후, $^1H$와 $^{13}C-NMR$ MS, HMQC 및 COSY spectrum(500 MHz $CDCl_3$) 분석을 통하여 분자량 328의 decursin임을 구조 동정하였다. 정제된 decursin을 MCF-7에 24시간 처리한 결과 $20{\mu}g/mL$ 이상에서 농도 의존적으로 암세포주의 성장을 억제하였으며, SW480, HepG2, MCF-7및 293과 같은 인체 암세포주에 대한 decurcin의 $IC_{50}$값은 각각 31.04, 27.24, 20.45 및 $33.60{\mu}g/mL$로 나타나, decursin은 인체 정상세포주인 293세포를 포함한 다른 세포에 비하여 MCF-7에 대하여 가장 높은 성장억제 활성을 보여주었다. MCF-7세포에 decursin을 처리한 결과 대조군에서 균일한 핵의 형태가 관찰된 것에 반해 처리군의 경우는 핵의 응축과 apoptotic body를 보였고, DNA 절편이 관찰되었다. 이 결과는 당귀에서 분리한 decursin은 MCF-7세포에서 apoptosis를 통하여 세포의 성장을 억제하는 것을 암시한다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1499-1506
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• 2016

Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.487-493
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• 2018

Cluster of differentiation 44 (CD44), a cell surface receptor for hyaluronic acid (HA), is involved in aggressive cancer phenotypes. Herein, we investigated the role of the CD44 standard isoform (CD44s) in hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) regulation using MCF7 overexpressing CD44s (pCD44s-MCF7). When pCD44s-MCF7 was incubated under hypoxia, levels of $HIF-1{\alpha}$, vascular endothelial growth factor, and the $HIF-1{\alpha}$ response element-derived luciferase activity were significantly increased compared to those in the control MCF7. Incubation of pCD44s-MCF7 cells with HA further increased $HIF-1{\alpha}$ accumulation, and the silencing of CD44s attenuated $HIF-1{\alpha}$ elevation, which verifies the role of CD44s in $HIF-1{\alpha}$ regulation. In addition, the levels of phosphorylated extracellular signal-regulated kinase (ERK) was higher in hypoxic pCD44s-MCF7 cells, and $HIF-1{\alpha}$ accumulation was diminished by the pharmacological inhibitors of ERK. CD44s-mediated $HIF-1{\alpha}$ augmentation resulted in two functional outcomes. First, pCD44s-MCF7 cells showed facilitated cell motility under hypoxia via the upregulation of proteins associated with epithelial-mesenchymal transition, such as SNAIL1 and ZEB1. Second, pCD44s-MCF7 cells exhibited higher levels of glycolytic proteins, such as glucose transporter-1, and produced higher levels of lactate under hypoxa. As a consequence of the enhanced glycolytic adaptation to hypoxia, pCD44s-MCF7 cells exhibited a higher rate of cell survival under hypoxia than that of the control MCF7, and glucose deprivation abolished these differential responses of the two cell lines. Taken together, these results suggest that CD44s activates hypoxia-inducible $HIF-1{\alpha}$ signaling via ERK pathway, and the $CD44s-ERK-HIF-1{\alpha}$ pathway is involved in facilitated cancer cell viability and motility under hypoxic conditions.

• 생명과학회지
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• 제21권9호
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• pp.1288-1294
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• 2011

식물에서 추출한 파이토케미컬은 암세포의 여러 신호전달 기작에 관여함으로써 apoptosis를 유도한다. 본 연구에서는 파이토케미컬의 한 종류인 레스베라트롤을 MCF-7 세포에 처리함으로써 암세포의 증식 억제와 apoptosis 유도 효과를 알아보았고, 이러한 효과가 암세포의 성장과 증식에 관여하는 단백질인 mTOR와 COX-2의 발현 양상에 어떠한 영향을 미치는지 알아보고자 하였다. 그 결과 MCF-7 세포에 레스베라트롤을 처리했을 때 농도가 증가함에 따라 암세포의 생존률이 감소하였고, Hoechst 33342를 이용한 chromatin 염색과 Annexin V-propodium iodide staning을 통하여 암세포의 세포증식 효과가 apoptosis에 의해 유도된 것임을 알 수 있었다. MCF-7 세포에 레스베라트롤을 처리했을 때 mTOR 및 COX-2의 발현 양상을 확인하기 위해 Western blotting을 실시한 결과, 레스베라트롤의 농도가 높아짐에 따라 mTOR 및 COX-2의 발현이 감소함을 확인 하였다. 이와 같은 결과는 MCF-7 유방암 세포에서 레스베라트롤에 의한 암세포의 증식 억제 및 apoptosis 유도가 mTOR 신호경로 저해를 통한 COX-2의 발현을 감소시킴으로써 나타나는 것으로 보인다.

• Environmental Analysis Health and Toxicology
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• 제27권
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• pp.10.1-10.7
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• 2012

Objectives: In recent years, a number of structurally diverse Histone deacetylase (HDAC) inhibitors have been identified and these HDAC inhibitors induce growth arrest, differentiation and/or apoptosis of cancer cells in vitro and in vivo. This study aimed at investigating the antitumor activity of newly synthesized HDAC inhibitor, 3-(4-dimethylamino phenyl)-N-hydroxy-2-propenamide (IN-2001) using human breast cancer cells. Methods: We have synthesized a new HDAC inhibitor, IN-2001, and cell proliferation inhibition assay with this chemical in estrogen receptor-positive human breast cancer MCF-7 cells. Cell cycle analysis on MCF-7 cells treated with IN-2001 was carried out by flow cytometry and gene expression was measured by RT-PCR. Results: In MCF-7 cells IN-2001 showed remarkable anti-proliferative effects in a dose- and time-dependent manner. In MCF-7 cells, IN-2001 showed a more potent growth inhibitory effect than that of suberoylanilide hydroxamic acid. These growth inhibitory effects were related to the cell cycle arrest and induction of apoptosis. IN-2001 showed accumulation of cells at $G_2$/M phase and of the sub-$G_1$ population in a time-dependent manner, representing apoptotic cells. IN-2001-mediated cell cycle arrest was associated with HDAC inhibitor-mediated induction of CDK inhibitor expression. In MCF-7 cells, IN-2001 significantly increased $p21^{WAF1}$ expression. Conclusions: In summary, cyclin-dependent kinase (CDK) induced growth inhibition, possibly through modulation of cell cycle and apoptosis regulatory proteins, such as CDK inhibitors, and cyclins. Taken together, these results provide an insight into the utility of HDAC inhibitors as a novel chemotherapeutic regime for hormone-sensitive and insensitive breast cancer.

• Natural Product Sciences
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• 제22권4호
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• pp.293-298
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• 2016

Plant-derived triterpenoids commonly possesses biological properties such as anti-inflammatory, antimicrobial, anti-viral and anti-cancer. Luvunga scandens is one of the plant that produced triterpenoids. The aims of the study was to analyze cell cycle profile and to determine the expression of p53 unregulated modulator of apoptosis (PUMA), caspase-8 and caspase-9 genes at mRNA level in MCF-7 cell line treated with two triterpenoids, flindissol (1) and 3-oxotirucalla-7,24-dien-21-oic-acid (2) isolated from L. scandens. The compounds were tested for cell cycle analysis using flow cytometer and mRNA expression level using quantitative RT-PCR. The number of MCF-7 cells population which distributed in Sub G1 phase after treated with compound 1 and 2 were 7.7 and 9.3% respectively. The evaluation of the expression of genes showed that both compounds exhibited high level of expression of PUMA, caspase-8 and caspase-9 as normalized to ${\beta}-actin$ via activation of those genes. In summary, the isolated compounds of L. scandens plant showed promising anticancer properties in MCF-7 cell lines.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8113-8117
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• 2016

Glehnia littoralis (GL) is widely used as an oriental medicine for cough, fever, stroke and other disease conditions. However, the anti-cancer properties of GL on MCF-7 human breast cancer cells have not been investigated. In order to elucidate anti-cancer properties and underlying cell death mechanisms, MCF-7cells ($5{\times}10^4/well$) were treated with Glehnia littoralis root extract at 0-400 ug/ml. A hot water extract of GL root inhibited the proliferation of MCF-7 cells in a dose-dependent manner. Analysis of the cell cycle after treatment of MCF-7 cells with increasing concentrations of GL root extract for 24 hours showed significant cell cycle arrest in the G1 phase. RT-PCR and Western blot analysis both revealed that GL root extract significantly increased the expression of p21 and p27 with an accompanying decrease in both CDK4 and cyclin D1. Our reuslts indicated that GL root extract arrested the proliferation of MCF-7 cells in G1 phase through inhibition of CDK4 and cyclin D1 via increased induction of p21 and p27. In summary, the current study showed that GL could serve as a potential source of chemotherapeutic or chemopreventative agents against human breast cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1511-1515
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• 2014

Background and Aims: To explore the molecular mechanisms of miR-886-5p in breast cancer., we examined roles in inhibiting growth and migration of MCF-7 cells. Methods: MiR-886-5p mimics and inhibitors were used to express or inhibit MiR-886-5p, respectively, and MTT and clone formation assays were used to determine the survival and proliferation. Hoechst 33342/ PI double staining was applied to detect apoptosis. The expression of caspase-3, caspase-8, caspase-9, MT1-MMP, VEGF-C and VEGF-D was detected by Western blotting, and the levels of MMP2 and MMP9 secreted from MCF-7 cells were assessed by ELISA. MCF-7 cell migration was determined by wound healing and Transwell assays. Results: We found that the growth of MCF-7 cells was inhibited upon decreasing miR-886-5p levels. Inhibiting miR-866-5p also significantly induced apoptosis and decreased the migratory capacity of these cells. The expression of VEGF-C, VEGF-D, MT1-MMP, MMP2, and MMP9 was also found to be decreased as compared to controls. Conclusions: Our data show that downregulation of miR-886-5p expression in MCF-7 cells could significantly inhibit cell growth and migration. This might imply that inhibiting miR-886-5p could be a therapeutic strategy in breast cancer.

• 대한한방부인과학회지
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• 제28권2호
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• pp.26-39
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• 2015

Objectives : Dendropanax morbifera is known as a tree that has been used in traditional medicine for various diseases. However, its biological activities in cancer have not yet been clearly elucidated. In this study, we investigated the anti-cancer effects of water extract of Dendropanax morbifera (DP) on 2 human breast cancer cell lines (estrogen dependent MCF-7 and estrogen independent MDA-MB-231). Methods : The MTT assay and flow cytometry were used to assess cell proliferation, along with cell cycle analysis. Nitric oxide production was detected by Griess assay. The expression of apoptosis related gene was assessed by quantitative real-time PCR. Results : Our data revealed that DP inhibits the cell growth in a dose dependent manner (0, 50, 100, 250, and 500 μg/ml) of both estrogen independent MDA-MB-231 and estrogen dependent MCF-7 breast cancer cells. Also, LPS induced nitric oxide production was significantly reduced by DP. Cell cycle analysis showed an increased G1 phase in the MCF-7 cell and G2/M phase in the MDA-MB-231 cell. DP decreased mRNA expression of apoptotic suppressor gene Bcl-xL, and increased mRNA expression of pro-apoptotic genes. DP increased mRNA expression of p21, and Rip1 in both cell. And DP decreased mRNA expression of survivin in the MCF-7 cell. Conclusions : Taken together, these results indicate that DP extract are source of anti-cancer potential and could be developed botanical drug.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5719-5723
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• 2013

Pharmacological and preventive properties of Petroselinum sativum seed extracts are well known, but the anticancer activity of alcoholic extracts and oil of Petroselinum sativum seeds on human breast cancer cells have not been explored so far. Therefore, the present study was designed to investigate the cytotoxic activities of these extracts against MCF-7 cells. Cells were exposed to 10 to $1000{\mu}g/ml$ of alcoholic seed extract (PSA) and seed oil (PSO) of Petroselinum sativum for 24 h. Post-treatment, percent cell viability was studied by 3-(4, 5-dimethylthiazol-2yl)-2, 5-biphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed that PSA and PSO significantly reduced cell viability, and altered the cellular morphology of MCF-7 cells in a concentration dependent manner. Concentrations of $50{\mu}g/ml$ and above of PSA and $100{\mu}g/ml$ and above of PSO were found to be cytotoxic in MCF-7 cells. Cell viability at 50, 100, 250, 500 and $1000{\mu}g/ml$ of PSA was recorded as 81%, 57%, 33%, 8% and 5%, respectively, whereas at 100, 250, 500, and $1000{\mu}g/ml$ of PSO values were 90%, 78%, 62%, and 8%, respectively by MTT assay. MCF-7 cells exposed to 250, 500 and $1000{\mu}g/ml$ of PSA and PSO lost their typical morphology and appeared smaller in size. The data revealed that the treatment with PSA and PSO of Petroselinum sativum induced cell death in MCF-7 cells.