Cytotoxic and Apoptotic Properties of the Flavonoid-rich Ethyl Acetate Fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells

Introduction@#Breast cancer is the most common cancer among women in the Philippines and about 3 in every 100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer. @*Objectives@#This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines. @*Methods@#Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining. @*Results@#The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses. @*Conclusion@#The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.

Isobavachalcone induces cell death through multiple pathways in human breast cancer MCF-7 cells / 南方医科大学学报

OBJECTIVE@#To explore the effects of isobavachalcone (IBC) on cell death of human breast cancer MCF-7 cells and explore the possible mechanism.@*METHODS@#MCF-7 cells were treated with different concentrations of IBC, and the changes in cell proliferation were assessed using MTT assay. Apoptosis of MCF-7 cells following treatment with 10, 20, and 40 μmol/L IBC was analyzed using flow cytometry with annexin V-FITC/PI double staining and fluorescence microscopy, and the expressions of apoptosis- and autophagy-related proteins (Bax, Bcl-2, Akt, p-Akt, p62, and LC3) were detected with Western blotting. Electron microscopy was used to observe the changes in submicrostructure of the cells following treatment with 40 μmol/L IBC. JC-1 assay kit, ATP assay kit, and reactive oxygen species (ROS) kit were used to determine the effect of IBC on mitochondrial function of the cells.@*RESULTS@#MTT assay showed that IBC significantly inhibited the proliferation of MCF-7 cells in a concentration- and time-dependent manner, with IC50 values of 38.46, 31.31, and 28.26 μmol/L at 24, 48, and 72 h, respectively. IBC also concentration-dependently induced apoptosis of MCF-7 cells. IBC-induced cell death was inhibited by z-VAD-fmk, a caspase inhibitor (P < 0.05), but not by the necroptosis inhibitor necrostatin-1 (Nec-1). Western blotting showed that IBC-induced MCF-7 cell apoptosis by increasing Bax expression and down-regulating the expressions of Bcl-2, Akt and p-Akt-473 (all P < 0.05). With the increase of IBC concentration, the expression of autophagy-related protein p62 and the LC3-II/I ratio increased progressively. Electron microscopy revealed the presence of autophagic bodies in IBC-treated MCF-7 cells. IBC treatment also resulted in decreased mitochondrial membrane potential and intracellular ATP level and increased ROS accumulation in MCF-7 cells (P < 0.05).@*CONCLUSION@#IBC is capable of inducing both apoptosis and autophagy in MCF-7 cells, suggesting the potential value of IBC as a lead compound in the development of anti-breast cancer agents.

In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

Exploration of organoid in breast cancer related research / 生物工程学报

Breast cancer is the most common cancer in women. At present, the in vivo model and traditional cell culture are mainly used in breast cancer researches. However, as high as 90% clinical trials are failed for drugs explored by the above two methods, due to the inherent species differences between humans and animals, as well as the differences in the tissue structure between organs and cells. Therefore, organoid three-dimensional culture is emerging. As a new tumor research model, organoid, a three-dimensional cell complex with spatial structure, has broad application prospects, such as precision medicine, organ transplantation, establishment of refractory disease model, gene therapy and drug research and development. Therefore, organoid is considered as one of the ideal carriers for life science research in the future. Breast cancer, a heterogeneous disease with complex phenotypes, has a low survival rate. Breast cancer organoid can reproduce many key features of human breast cancer, thus, the construction of organoid biological library of breast cancer will provide a new platform for studying the occurrence, development, metastasis and drug resistance mechanism of breast cancer. In this review, we systematically introduce the culture conditions of organoids and their application in breast cancer related research, and the application prospect of organoids.

A nanoemulsion-based delivery system for imatinib and in vitro anticancer efficacy

A self-nanoemulsifying drug delivery system (SNEDDS) composed of ethyl oleate, Tween 80 and polyethylene glycol 600 was prepared as a new route to improve the efficacy of imatinib. The drug-loaded SNEDDS formed nanodroplets of ethyl oleate stabilized by Tween 80 and polyethylene glycol 600 with a diameter of 81.0±9.5 nm. The nanoemulsion-based delivery system was stable for at least two months, with entrapment efficiency and loading capacity of 16.4±0.1 and 48.3±0.2%, respectively. Imatinib-loaded SNEDDS was evaluated for the drug release profiles, and its effectiveness against MCF-7 cell line was investigated. IC50 values for the imatinib-loaded SNEDDS and an imatinib aqueous solution were 3.1 and 6.5 µg mL-1, respectively.

Cytotoxic and Apoptotic properties of the Flavonoid-rich Ethyl Acetate fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells

Introduction@#Breast cancer is the most common cancer among women in the Philippines and about 3 in every 100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer. @*Objectives@#This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines. @*Methods@#Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining. @*Results@#The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses. @*Conclusion@#The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of S. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.

Enhancing effect of Lupeol on migration and invasion abilities of ST3Gaim-silenced MDA-MB-231 cells / 解剖学报

Objective In this study, we expored the enhancing effect of Lupeol on migration and invasion abilities of ST3Gal III-silenced MDA-MB-231 cells. Methods Human breast cancer cell line ST3Gal III -silenced MDA-MB-231 was cultured in vitro. The cell adhesion, Transwell and woud healing test were utilized to test the effect of Lupeol on ST3GalM-silenced MDA-MB-231 cells. The expression of matrix metalloproteinase( MMP)-2, -9, phosphatidylinositol 3-kinase/protein kinase B/nuclear factor κB ( PI3K/Akt/NF-KB) signaling pathway were examined by Western blotting. Results No significant influence of the decreased expression of ST3Gal III and Lupeol ( 5μmol/L) on proliferation and apoptosis of MDA-MB-231 cells were found. Lupeol inhibited the migration and invasion of ST3Gal III -silenced MDA-MB-231 cells in vilro( P<0. 05). Furthermore, the expression of NF-κB p65, p-Akt, Akt, p85, MMP-2 and MMP-9 levels were significantly down-regulated. Conclusion These observations suggest that Lupeol may inhibit the abilities of migration and invasion of ST3Gal III-silenced MDA-MB-231 cells in vitro by inhibiting the protein expression of MMP-2, -9 and effect the signaling pathway of PI3K/Akt/NF-κB.

Regulatory effects of glabridin and quercetin on energy metabolism of breast cancer cells / 中国中药杂志

It is reported that energy metabolism is the core feature of tumor cells. This study is aimed to investigate the regulatory effect of two flavonoids( glabridin and quercetin) on energy supply and glycolysis of breast cancer cells,and provide reference for developing some anticancer herbal drugs with the function of regulating tumor energy metabolism. Based on the characteristics of each pathway during energy metabolism,in the present study,the triple negative breast cancer tumor cells( MDA-MB-231) were selected to investigate the effects of glabridin and quercetin on the energy metabolism of breast cancer cells and discuss the possible mechanisms from the following five potential targets: glucose uptake,protein expression of glucose transporter 1( GLUT1),adenosine triphosphate( ATP) level,lactate dehydrogenase( LDH) activity,and lactic acid( LD) concentration. The results showed that both quercetin and glabridin could decrease the glucose uptake capacity of breast cancer cells by down-regulating the protein expression of GLUT1. Quercetin had no significant effect on LDH activity and LD concentration; it did not affect the glycolysis process,but increased the intracellular ATP level. Glabridin decreased the activity of LDH and reduced LD concentration,thereby inhibiting the glycolysis metabolism of breast cancer cells. Therefore,both quercetin and glabridin can regulate the energy metabolism of breast cancer cells and can be used as potential anticancer agents or anti-cancer adjuvants.

Synergistic Effect of Flavonoids from Artocarpus heterophyllus Heartwoods on Anticancer Activity of Cisplatin Against H460 and MCF-7 Cell Lines

Artocarpus heterophyllus has been used as traditional medicine. This plant is one of the sources of flavonoid. Flavonoid compounds possessed a wide range of biological properties including anticancer. This study was performed to investigate the cytotoxic effect of flavonoids from A. heterophyllus on H460 and MCF-7 cell lines. The interaction of flavonoids and cisplatin against tested cancer cells was also evaluated. MTT assay was used to determine the cytotoxic effect of flavonoid. Isobologram analysis was selected to evaluate the synergistic effect between flavonoid and cisplatin, their interaction was then confirmed using AO/PI staining method. Amongst of flavonoid compounds, artocarpin exhibited strong cytotoxic effect on both MCF-7 and H460 cell lines with IC₅₀ values of 12.53 µg/mL (28.73 µM) and 9.77 µg/mL (22.40 µM), respectively. This compound enhanced anticancer activity of cisplatin against H460 and MCF-7. The combination produced a synergistic effect on H460 and MCF-7 cell lines with a combination index (CI) values of 0.2 and 0.18, respectively. The AO/PI stained demonstrated that the combination of artocarpin and cisplatin caused morphological changes that indicated apoptosis. Moreover, artocarpanone also significantly increased cytotoxic effect of cisplatin compared to its single concentration with CI below than 1. This result suggested the potency of flavonoid named artocarpin to enhance the anticancer activity of cisplatin on H460 and MCF-7 cell lines.

Effect of hypoxia on epithelial growth factor receptor expression and cell apoptosis of breast cancer and cervical cancer xenografts in nude mice / 中华放射肿瘤学杂志

Objective To observe the effect of hypoxia on the expression of epithelial growth factor receptor (EGFR) and cell apoptosis of breast and cervical cancer xenografts in nude mouse models.Methods The nude mouse models with MCF-7 and HeLa xenografts were established.The degree of hypoxia and EGFR expression were observed by confocal microscopy.The influence of EGFR expression on cell apoptosis under hypoxia was observed by TUNEL assay.Results EGFR expression was either up-regulated or down-regulated in the MCF-7 and HeLa cells with high degree of hypoxia.Furthermore,the degree of apoptosis was reduced in tumor tissues with high EGFR expression compared with that in those with low expression of EGFR.Conclusion The hypoxia in MCF-7 and HeLa cells exerts heterogeneous effect on EGFR expression.Under hypoxic conditions,EGFR exoression is negatively correlated with cell apoptosis.

Value of the intraoperative cytology examination of sentinel lymph node in breast cancer / Valor do exame citológico intraoperatório do linfonodo sentinela no câncer de mama

Objective: Evaluate the value of imprint cytology in the intraoperative analysis of sentinel lymph node (SLN) in patients with breast cancer. Methods: An agreement study for the evaluation of the imprint cytology technique as a diagnostic test for intraoperative SLN among patients diagnosed with breast cancer from January 2007 to January 2017. Results: We studied 210 cases of breast cancer patients submitted to intraoperative sentinel node imprint cytology, aged between 24 and 86 years (mean age 59 years and median age 60 years). The sensitivity of the intraoperative study was 58.3% (95%CI 46.1­69.8%) and the specificity was 97.8% (95%CI 93.8­99.5). The positive predictive value (PPV) was 93.3% (95%CI 81.7­98.6) and the negative predictive value (NPV) was 81.8% (95%CI 75.1­87.4). From the analyzed variables, the presence of macrometastasis was the only one that significantly increased the sensitivity of the imprint to 73.2% (95%CI 59.7­84.2), while micrometastasis presented a sensitivity of only 6.3% (95%CI 0.2­30.2). Conclusion: The use of imprint cytology in the intraoperative SLN study showed good accuracy in predicting axillary status. However, the surgeon and pathologist are fully aware of the set of clinical and histological variables that can influence the sensitivity of the method.

Objetivo: Avaliar o valor do imprint citológico na análise intraoperatória do linfonodo sentinela (LS) em pacientes com câncer de mama. Métodos: Estudo de concordância para avaliação da técnica do imprint citológico como teste diagnóstico do LS no intraoperatório, entre pacientes com diagnóstico de câncer de mama, no período de janeiro de 2007 a janeiro de 2017. Resultados: Foram estudados 210 casos de pacientes com câncer de mama submetidas à citologia de impressão (IC) do linfonodo sentinela no intraoperatório, com idade entre 24 e 86 anos (média de 59 anos e mediana de 60 anos). A sensibilidade do estudo intraoperatório foi de 58,3% (IC95% 46,1­69,8) e a especificidade de 97,8% (IC95% 93,8­99,5). O valor preditivo positivo (VPP) foi de 93,3% (IC95% 81,7­98,6) e o valor preditivo negativo (VPN) de 81,8% (IC95% 75,1­87,4%). Das variáveis analisadas, a presença de macrometástase foi a única que aumentou significativamente a sensibilidade do imprint para 73,2% (IC95%, 59,7­84,2%), enquanto na micrometástase apresentou sensibilidade de apenas 6,3% (IC95% 0,2­30,2). Conclusão: A utilização do imprint citológico no estudo intraoperatório do LS apresentou boa acurácia na previsão do status axilar. Entretanto, é importante o pleno conhecimento, pelo cirurgião e patologista, do conjunto de variáveis clínicas e histológicas que podem influenciar a sensibilidade do método

Effect of matrine on tumor growth and inflammatory factors and immune function in Wistar rat with breast cancer / 中国应用生理学杂志

OBJECTIVE@#To study the effect of matrine on tumor growth, inflammatory factors and immune function in Wistar rat with breast cancer.@*METHODS@#Sixty female Wistar rats were randomly divided into control group (=10) and the modeling group of breast cancer cell tumor-bearing rat (=50), then the rats in modeling group were randomly divided into five groups (=10):vehicle group, matrine low dose group (50 mg/kg), medium dose group (100 mg/kg), high dose group (200 mg/kg), and lentinan group (200 mg/kg). Except the control group, each rat in the other groups was subcutaneously inoculated 0.4 ml Walker 256 breast cancer cell suspension (5×10 cells/ml) in the right axillary. Each group was treated with corresponding drug by ig administration (10 ml/kg body weight) twice a day for 14 days. After 14 days, the blood sample was collected from ventral aorta, the tumor was removed and weighed to calculate tumor inhibitory rate. The levels of interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth factor-β (TGF-β), CD3, CD4, CD8, IgG, IgM, IgA in peripheral blood were determined.@*RESULTS@#The mean tumor weight of matrine low-dose, medium-dose, high-dose groups and lentinan group were (4.99±0.93) g, (4.52±0.92) g, (4.22±1.18) g and (4.52±0.92) g respectively, which were significantly lower than that in model group. There was no statistical difference on the mean tumor weight among matrine groups and lentinan group (>0.05). After the drug intervention, the tumor inhibitory rates of matrine low-dose, medium dose, high-dose groups and lentinan group were 24.6%, 31.7%, 36.3%, and 27.9% respectively, there was no statistical difference among the four groups. The levels of IL-2, IFN-γ, CD8+ in vehicle group were lower than those of control group obviously (<0.01), however, the levels of IL-6, IL-10, TGF-β, CD3, CD4, IgG, IgM, IgA were higher significantly than those of control group (<0.01). The levels of IL-2, IFN-γ, CD8 in matrine low-dose, medium dose, high-dose groups and lentinan group were higher than those of vehicle group obviously (<0.01, <0.05); while the levels of IL-6, IL-10, TGF-β, CD3, CD4, IgG, IgM, IgA were lower than those of model group markedly (<0.01, <0.05). The levels of IgM and IgA in matrine low-dose and medium-dose groups were higher than those of lentinan group obviously (<0.01, <0.05); the levels of IL-2, IFN-γ and IgA in matrine high-dose group were higher than those of lentinan group obviously (<0.01, <0.05); while the levels of IFN-γ in matrine low-dose group were lower than those of lentinan group markedly (<0.05); the levels of IL-10 and CD4 in matrine high-dose group were lower than those of lentinan group markedly (<0.01, <0.05).@*CONCLUSIONS@#Matrine has an obvious antitumor action which is related to its ability to enhance cellular and humoral immunity, reduce inflammatory reaction.

Preparation of Argininate Betulinic Acid and Its Effect of the Proliferation of Triple-negative Human Breast Cancer Cell MDA-MB-231 / 中国药房

OBJECTIVE:To prepare argininate betulinic acid,and to investigate the effect of the proliferation of triple-negative human breast cancer cell MDA-MB-231. METHODS:By using argininate as the solubilization carrier,argininate betulinic acid was prepared by co-grinding equal molar ratio of betulinic acid and argininate. The argininate betulinic acid was characterized with powder X-ray diffractometry,infrared spectroscopy and differential scanning calorimetry. The solubility of betulinic acid and argininate betulinic acid were compared. MTT method was used to assay the effects of 15,30,60,120 μ g/mL betulinic acid, argininate betulinic acid and 5-FU on the proliferation of MDA-MB-231 cell. RESULTS:Prepared argininate betulinic acid was a new phase which was different from the physical mixing of argininate and betulinic acid,among which carboxyl group of betulinic acid and amino group of argininate formed as a salt,and the salt had no obvious melting peak. Betulinic acid was almost insoluble in water. The solubility of betulinic acid in argininate betulinic acid aqueous solution was 50.72 μg/mL. Compared with betulinic acid,the inhibitory rate of argininate betulinic acid on the growth of MDA-MB-231 cell was increased significantly(P<0.05), there was no statistical significance between its effect and 5-FU(P>0.05). CONCLUSIONS:Argininate betulinic acid with good solubility is prepared successfully,and can inhibit the proliferation of MDA-MB-231 cell.

The Suppressive Effect of Carthamin Yellow on the Proliferation and Migration of Human Breast Cancer Cells and Its Related Molecular Mechanisms / 昆明医科大学学报

Objective To study the effect of Carthamin Yellow (CY) on cell proliferation, apoptosis, migration and invasion ability of breast cancer and its related molecular mechanisms. Methods CCK-8 assay was used to detect cell viability of MDA-MB-231 human breast cancer cells by different concentrations of CY at different time;flow cytometry was used to test the apoptosis rate of MDA-MB-231 cells treated by different concentrations of CY and transwell assay was used to investigate the effect of various concentrations of CY on MDA-MB-231 cell migration and invasion.After the intervention of different concentrations of CY on MDA-MB-231 cells, apoptosis-related protein Cleaved-Caspase-3, survival protein p-Akt and metastasis-related protein MMP2 were detected by western blot. Results (1) CY could inhibit the proliferation of MDA-MB-231 cells in a dose-and-time-dependent manner. (2) CY significantly promoted the apoptosis of breast cancer cells ( <0.01) . (3) CY could decrease the expression of p-Akt and increase the expression of Cleaved-Caspase-3. (4) CY impaired migration and invasion of MDA-MB-231 cells ( <0.01), and can inhibit the expression of MMP2. Conclusion CY could promote the apoptosis of breast cancer cells through activation of apoptosis signaling, and can inhibit breast cancer cell metastasis by suppressing MMP2. And CY may be a potential therapeutic drug for human breast cancer.

Synthesis, crystal structure, and anti-breast cancer activity of a novel metal-porphyrinic complex [YK(TCPP)(OH)2·(solvents)x]

A novel heterometallic metal-porphyrinic framework (MPFs) built from Y and K ions as nods and meso-tetra(4-carboxyphenyl)porphyrin as linkers has been successfully synthesized and characterized. The single crystal X-ray diffraction indicated that this complex 1 exhibited a bilayered architecture of the porphyrins, which is seldom seen in MPFs. In addition, in vitro anticancer activity of complex 1 on three human breast cancer cells (BT474, SKBr-3 and ZR-75-30) was further determined.

Effect of dihydromyricetin on proliferation and apoptosis of breast cancer MCF-7 cells / 国际检验医学杂志

Objective To investigate the effect of dihydromyricetin on proliferation and apoptosis of breast cancer MCF-7 cells.Methods From March 2014 to February 2015,breast cancer MCF-7 cells were treated with 99% pure DMY as an inhibitor.MTT assay,flow cytometry and immunocytochemistry were used to analyze the proliferation,apoptosis and protein expression of breast cancer cell MCF-7.Results When the DMY concentration was higher than 20 μg/mL,the inhibitory effect appeared,but not good.When 40 and 80 μg/mL DMY were used,the proliferation of MCF-6 cells were significantly inhibited,and have different degrees of sensitivity to it.When DMY was 80 μg/mL,the IC50 was 226.9 μg/mL.The inhibition rate and IC50 were compared with 0 μg/mL DMY,there was significant difference(P 50%,especially in DMY with 80 μg/mL,the positive rate was 10.00%.Compared with 0 μg/mL DMY,the difference was significant(P<0.05).Conclusion The use of dihydromyricetin in breast cancer patients can effectively inhibit the rapid increase of cancer cells,accelerate apoptosis,slow down the patient′s condition,the effect is outstanding.

Impact of TLR5 and NLRC4 activation on proliferation of different breast cancer cell lines / 中国免疫学杂志

Objective:To explore the impact of TLR5 and NLRC4 activation on the proliferation of different breast cancer cell lines,MCF-7 and MDA-MB-23 i.Methods:Induction,expression,purification and identification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97 (unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4).Using different concentration of recombinant flagellin to stimulate MCF-7 and MDA-MB-231 cell lines,72 h later,the proliferation of tumor cells were detected with CCK8.We also used soft AGAR forming experiments to detect the inhibition ratio of recombinant flagellin on breast cancer cell lines.Briefly,1 000 cells were plated in the 6-well plate,then stimulated with 1 μg/ml recombinant flagellin,14 days later,the number of cloning were counted after crystal violet staining.Results:After stimulation with four recombinant flagellins at the concentration of 0.1 μ,g/ml,the inhibition ratio on MCF-7 reached 30％,and FliC△90-97 were dose-dependent on the inhibition of MCF-7 proliferation.At the concentration of 1 μg/ml,FliC-L3A which only activated TLR5 showed stronger inhibition ratio than FliC.FliC△90-97:L3A which did not activate both TLR5 and NLRC4 also inhibited the proliferation of MCF-7.After adding transfection reagent,four recombinant flagellins showed inhibition effect on MDA-MB-231.Conclusion:Flagellin can inhibit the proliferation of MCF-7 and MDA-MB-231,and the mechanism of inhibition on the proliferation were not TLR5 and NLRC4 pathway dependent.There might exist new mechanisms to explain this phenomenon.

Anti-cancer by baicalein combined with U0126 on human breast cancer in vitro / 中国免疫学杂志

Objective:To investigate the effects and mechanisms of anti-cancer by bacailein combined with U0126 on human breast cancer in vitro. Methods: The human breast cancer cell line MCF-7 was treated by baicalein,U0126 and baicalein combined with U0126 respectively. CCK8 assay measured cell proliferation of MCF-7;flow cytometry tested the cell cycle and apoptosis of MCF-7;microscopy observed the amount;TUNEL assay evaluated the apoptosis of MCF-7;Western blot detected the protein level of proliferation and apoptosis related protein;scratch assay measured the ability of migration. Results: Human breast cancer cell line MCF-7 was treated by baicalein or U0126 at different concentration for 24 h, CCK8 assay suggested that both of them can dramatically inhibit MCF-7 proliferation in a dose-dependent way (P<0. 05). Compared to the blank and DMSO groups,the human breast cancer cell line MCF-7 was treated with baicalein for 24 h,the cellular rate at G0-G1 phase increased a lot (91%) (P<0. 05),while the cellular rate at S phase reduced dramatically (P<0. 05),cell apoptosis increased dramatically by microscopy and TUNEL assay(P<0. 05),the level of ERK1/2,CyclinD1 and JNK reduced quickly (P<0. 05). Compared to the baicalein group,MCF-7 was treated by baicalein combined with U0126,the cellular rate at S phase decreased remarkably (P<0. 05),apoptosis was much obvious (P<0. 05),the phosphorylation level of ERK1/2 and JNK reduced a lot (P<0. 05),and the proliferation accelerator CyclinD1 highly decreased (P<0. 05);the scratch assay demonstrated that cell migration was dramatically inhibited when MCF-7 was treated by 20 μmol/L baicalein ( P<0. 05 ) . Conclusion:Both of baicalein and U0126 can inhibit the proliferation and migration,induce the apoptosis of human breast cancer cell line MCF-7 through decreasing the level of ERK, JNK and CyclinD1. Baicalein and U0126 can provide some novel avenues to treat breast cancer in clinic.

Flavonoids isolated from Sinopodophylli Fructus and their bioactivities against human breast cancer cells / 中国天然药物

Four prenylated flavonoids compounds 1-4, named sinopodophyllines A-D, and a flavonoid glycoside (compound 13), sinopodophylliside A, together with 19 known compounds (compounds 5-12 and 14-24) were isolated from the fruits of Sinopodophyllum hexandrum. The structures of new compounds were elucidated by extensive spectroscopic analysis, including HRESIMS, 1D and 2D NMR. Compounds 1-6, 9-11, and 14-17 were tested for their cytotoxicity against human breast-cancer T47D, MCF-7 and MDA-MB-231 cells in vitro, and compounds 2, 5, 6, 10 and 11 showed significant cytotoxicity (IC values < 10 μmol·L) against T47D cells.

Effect of lupeol on migration and invasion of human breast cancer MDA-MB-231 cells and its mechanism / 药学学报

In this study, we examined the inhibitory effects of lupeol, an extract of Euphorbia fischerana Steud, on human breast cancer MDA-MB-231 cells migration and invasion. Lupeol was found to inhibit the invasion of MDA-MB-231 in the cell adhesion assay, transwell test and wound healing assay. The expression of cyclooxygenase-2(COX-2), matrix metalloproteinase-2(MMP-2), -9(MMP-9) and nuclear transcription factor-kappa B (NF-κB) p65 in breast cancer following treatment with different concentrations of lupeol was analyzed with Western blot. Lupeol inhibited the migration and invasion of MDA-MB-231 cells in a dosedependent manner in vitro (PκB p65 levels was significantly down-regulated. These observations suggest that lupeol can inhibit the abilities of invasion of MDA-MB-231 cells by inhibiting the protein expression of COX-2, MMP-2 and MMP-9. Its mechanism may be related to inhibition of the nuclear NF-κB signal pathway.