Study on the mechanism of curcumol inhibiting the proliferation of breast cancer cells T 47D / 中国药房

OBJECTIVE To study the mechanism of curcumol inhibiting the pro liferation of breast cancer cells T 47D. METHODS MTT assay was used to detect the inhibitory effects of different doses of curcumol （0，6.25，12.5，25，50，100 μg/mL）on the proliferation of T 47D cells. After treated with curcumol （12.5，25，50，100 μg/mL），the morphology of T 47D cells was observed by inverted phase contrast microscope. The cell cycle and the levels of reactive oxygen species （ROS）were detected by flow cytometry. Quantitative real-time PCR （qRT-PCR）was used to detect the expressions of proliferating cell nuclear antigen （PCNA），cell cycle regular p 21 and cyclin-dependent kinase 2（CDK2）mRNA. Western blot assay was used to detect the protein expression of CDK 2，CDK6，Cyclin D ，PCNA，nucler transcription factor E 2-related factor （Nrf2）and Kelch-like ECH associated protein 1（Keap1）. Breast cancer cells T 47D were divided into 2 groups，one group was given different doses of curcumol ，and another group was given curcumol 33 μg/mL for 6，12，24，48 h. After the optimal oxidation time and administration concentration were determined according to the results of the above two groups ，the blank control group ，N-acetylcysteine（NAC）group（ROS antioxidant NAC alone ），curcumol group （curcumol alone ），curcumol combined with NAC group （pretreatment with ROS antioxidant NAC ，and then adding into curcumol ）. Cell cycle and fluorescence intensity of ROS were detected. RESULTS Curcumol could significantly increase the inhibitory rate of the proliferation of T 47D cells （P＜0.05 or P＜0.01），and showed a certain dose and time dependent trend. Curcumol blocked the ， cycle in the G 1 phase and significantly increased the level of ROS （P＜0.05 or P＜0.01）；ROS antioxidant NAC could significantly reverse above inductive effect of curcumol （P＜ 0.01）. qRT-PCR showed that curcumol down-regulated the com expression of PCNA and CDK 2 mRNA and up-regulated the expression of p 21 mRNA（P＜0.05 or P＜0.01）. Western blot assay showed that curcumol significantly down-regulated the edu.cn protein expression of Keap 1，Nrf2，CDK2，CDK6 and Cyclin D（P＜0.05，P＜0.01）；ROS antioxidant NAC could reverse the down-regulation effects of curcumol on the expression of these proteins（P＜0.05 or P＜0.01）. CONCLUSIONS Curcumol may induce oxidative stress and cell arrest in G 1 phase to inhibit the proliferation of T 47D cells.

Cytotoxicity Studies Of An Ethnobotanically Selected Solanum Pubescens Willd. Against Selected Human Cancer Cell Lines

Objective: The present study was aimed to determine the cytotoxicity concentration (CTC50) of different extracts made from the leaf and stem bark of an ethno botanically selected S. pubescens against Human liver carcinoma (Hep G2), Human colon carcinoma (CaCo2) and Human breast cancer (T-47 D) cell lines. Methods: Ethnobotanical survey was done through interviewing traditional medicinal practitioners then a potential herbal plant was selected after a thorough literature survey and its identity was confirmed. The soxhlet extraction method was adopted using five different solvents from leaf and stem bark powders of the study plant and the CTC50 of all the extracts were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay method. Results: Among the different extracts, CTC50 values were significant for stem bark extracts made from methanol (24.51±0.08 µg/ml) against Hep G2, while leaf chloroform extract was promising (57.15±1.75 µg/ml) against CaCo2 and n-hexane extract of leaf exhibited significant value (20.27±1.52 µg/ml) against T-47 D cancer cell lines. Conclusion: The major findings of the present study clearly provides evidence that the leaf and stem bark of S. pubescens possesses the potential anticancer bioactive compound solasodine.

Effects of Stilbene Glucoside on the Proliferation and Estrogen Receptor Expression of Human Breast Cancer T-47D Cells / 中国药房

OBJECTIVE：To study the effects of stilbene glucoside （TSG）on the proliferation and estrogen receptor （ER）of human breast cancer T- 47D cells ，and to explore its estrogen-like effect and potential mechanism. METHODS ：Taking ER positive human breast cancer T- 47D cells as subjects ，using β-estradiol（β-E2，1×10－8 mol/L）as positive control ，CCK-8 assay was used to detect the cell proliferation after treated with different concentrations of TSG （1×10－8，1×10－7，1×10－6，1×10－5，1×10－4 mol/L）for 24，48，72 h；the cell proliferation rate was calculated. Western blotting assay and RT-PCR methods were adopted to detect the protein and mRNA expression of ER-α and ER-β in cells after treated with low，medium and high concentrations of TSG （1×10－8， 1×10－6，1×10－4 mol/L）for 48 h. RESULTS ：After treated with different concentrations of TSG for 24，48，72 h，the cell proliferation rate of each administration group at each time point （except for β-E2 group at 48 h）increased significantly ，compared with blank group ；those of TSG groups （1×10－5，1×10－6，1×10－7 mol/L）were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. After treated with low ，medium and high concentrations of TSG for 48 h，protein and mRNA expression of ER-α and ER-β in cells were increased significantly，compared with blank group （P＜0.05 or P＜0.01）；protein expression of ER-β in TSG low concentration group ，mRNA expression of ER-α in TSG groups as well as mRNA expression of ER-β in TSG low and high concentration groups were significantly higher than β-E2 group（P＜0.05 or P＜0.01）. CONCLUSIONS ：TSG can induce the in vitro proliferation of T- 47D cells and exert estrogen-like effects by promoting protein and mRNA expression of ER-α and ER-β， which is stronger than that of β-E2 at a certain concentration.

Effects of Processed Polygonum multiflorum Containing Serum on the Proliferation and the Expression of ER of Human Breast Cancer T- 47D Cells / 中国药房

OBJECTIVE： To study the effects of processed Polygonum multiflorum containing serum on the proliferation and the expression of estrogen receptor （ER） of human breast cancer T-47D cells， and to investigate its phytoestrogen （PE）-like effect. METHODS： Sexually immature SD rats were randomly divided into estradiol valerate （Ev） group （positive control， 0.12 mg/kg）， processed P. multiflorum low-dose and high-dose groups （0.75， 3 g/kg， by crude drug）， low-dose and high-dose processed P. multiflorum+Ev groups （same dose as single drug group）， with 10 rats in each group. Blank group was given constant volume of water intragastrically， and administration groups were given relevant medicine intragastrically； once day and night， for consecutive 4 days. Two hours after last administration， blank serum and containing serum were prepared. T-47D cells were also randomly divided into blank group， Ev group， low-dose and high-dose processed P. multiflorum groups， low-dose and high-dose processed P. multiflorum+Ev groups， and then were cultured in medium which contained 20％ blank serum or drug containing serum. CCK-8 assay was used to detect proliferation rate （PR）. Western blotting assay and RT-PCR were used to detect the protein and mRNA expression of ER-α and ER-β. RESULTS： Compared with blank group， PR of administration groups [each administration group （24 h）， other administration groups （48， 72 h） except for high-dose processed P. multiflorum+Ev group] were increased significantly； high-dose processed P. multiflorum group （72 h） was significantly higher than Ev group， and low-dose processed P. multiflorum+Ev group （72 h） was significantly higher than the same-dose processed P. multiflorum group； high-dose processed P. multiflorum+Ev group （72 h） was significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. Relative protein expression of ER-α in Ev group， high-dose processed P. multiflorum group and low-dose processed P. multiflorum+Ev group， relative mRNA expression of ER-α and protein expression of ER-β in administration groups， relative mRNA expression of ER-β in Ev group， low-dose processed P. multiflorum group and processed P. multiflorum+Ev groups were all increased significantly. Relative protein and mRNA expression of ER-α in Ev group were significantly higher than processed P. multiflorum groups and combination groups. Relative protein and mRNA expression of ER-β in Ev group were significantly lower than low-dose processed P. multiflorum+Ev group， but relative mRNA expression of ER-β was significantly higher than processed P. multiflorum groups and high-dose processed P. multiflorum+Ev group. Relative protein and mRNA expression of ER-α and ER-β in low-dose processed P. multiflorum+Ev group as well as relative mRNA expression of ER-β in high-dose processed P. multiflorum+Ev group were significantly higher than the same-dose processed P. multiflorum group. Relative protein and mRNA expression of ER-α in high-dose processed P. multiflorum+Ev group were significantly lower than the same-dose processed P. multiflorum group （P＜0.05 or P＜0.01）. CONCLUSIONS： The processed P. multiflorum containing serum can promote the proliferation of human breast cancer T-47D cells， and play the PE-like role through promoting protein and mRNA expression of ER-α and ER-β. However， the above effects are weaker than estrogen， and the combination of the two may antagonize the effect of estrogen.

Effects of acacetin on T47D cell proliferation / 中国药理学通报

Aim To investigate the effect of acacetin on cell proliferation and the influence of acacetin on estrogen receptor expression in vitro.Methods The proliferation rates and the cell cycle changes of acace-tin-treated T47D cells were measured by sulforhodam-ine B(SRB)assay and flow cytometry,respectively. Moreover,the mRNA expressions of estrogen receptor-alpha(ERα),estrogen receptor-beta(ERβ)and pro-liferating antigen(Ki67)were determined by quantita-tive real time PCR (qPCR).Western blot was em-ployed to detect the ERαand ERβprotein expression. Results Acacetin significantly promoted the prolifera-tion and increased the amount of cells arrested in S and G2 /M phase under the concentration of 0.001 ~1 0μmol·L -1 .Ki67 mRNA level and the ERαprotein level in T47D cells were remarkably upregulated after acacetin treatment.To clarify which estrogen receptors played a role in acacetin induced the proliferation of T47D cells,the combination treatment of acacetin and ERαinhibitor (MPP)/ERβ inhibitor (PHTPP) was employed.We found that MPP could reverse the cell proliferation,the cell arrested in S and G2 /M phase and the increased Ki67 mRNA level induced by acace-tin.PHTPP also alleviated the T47D cell proliferation induced by acacetin,whereas no significant changes were found in cell cycle and Ki67 mRNA level.Con-clusion Acacetin stimulates the cell proliferation of T47D cells in the concentration from 0.001 μmol · L -1 to 1 0 μmol·L -1 ,which is mainly mediated by ERα.

Effects of Active Constituents of Sinopodophylli Fructus on Cell Proliferation,Cell Cycle and Mitochon-drial Membrane Potential of Human Breast Cancer Cell / 中国药房

OBJECTIVE:To explore the effects and mechanism of extracts,active constituents and constituent combination of Sinopodophylli Fructus on cell proliferation of human breast cancer. METHODS:Acid phosphatase method was conducted to deter-mine the effects of 4 extracts [ethanol extract (Xc),petroleum ether extract from ethanol extract (Xp),ethyl acetate extract from ethanol extract (Xe),n-butanol extract from ethanol extract (Xz)],5 active constituents [podophyllotoxin (S1),deoxypodophyllo-toxin (S2),4-desmethyl deoxypodophyllotoxin (S3),8-isopentenyl kaempferol (S4),8,2′-diisoprenyl quercetin-3-methyl ether (S5)] and 3 active constituent combination [combination 1,S1-S2-S3-S4-S5 (2:4:1:4:32),Z1;combination 2,S2-S4 (1:1),Z2;combination 3,S3-S4(1:4),Z3] on the MDA-MB-231,MCF-7 cell proliferation;flow cytometry was adopted to detect the effects of above-mentioned samples on MDA-MB-231,MCF-7(T47D)cell cycle and mitochondrial membrane potential. RESULTS:The active constituent combination Z1 showed significant inhibitory effects on MDA-MB-231,MCF-7 cells,the half inhibitory concen-trations(IC50)were(0.27±0.2),(0.11±0.1)μg/mL;extracts Xc,Xp,Xe,active constituents S2,S4 and active constituent combi-nation Z2,Z3 showed relatively strong inhibitory effects on MDA-MB-231,MCF-7 (T47D) cell proliferation (IC50<15 μg/mL). Both extracts and active constituents can block MDA-MB-231,MCF-7 cell cycle in G2/M phase;all active constituents can block MDA-MB-231,T47D cell cycle in G0/G1 phase,and can reduce MDA-MB-231,T47D cell mitochondrial membrane potential. CONCLUSIONS:The active constituents and constituent combination of Sinopodophylli Fructus can inhibit cell proliferation of breast cancer by affecting cell cycle and mitochondrial mem-brane potential.

Cytotoxic activity and phytochemical standardization of Lunasia amara Blanco wood extract

Objective: To evaluate the cytotoxic activity of wood extracts of Lunasia amara Blanco (L. amara) and to perform further phytochemical standardization. Methods: The wood extracts of L. amara were assessed for cytotoxic activity by in vitro tetrazolium bromide (MTT) method against two human cancer cell lines, cervical cancer cells (HeLa) and breast cancer cells (T47D). Thin layer chromatography, Dragendorf, acetic anhydride-sulfuric acid and ferric chloride were used to detect alkaloids, steroids and polyphenols, respectively. Furthermore, quantitative determination of total alkaloid by ultra fast liquid chromatography-photodiode array detection using lunacrine as a marker compound was performed as well. Results: The ethyl acetate extract exhibited higher cell-growth inhibition than methanol and n-hexane extracts on HeLa and T47D cancer line cells with the IC50 of 71.15 and 79.04 mg/mL, respectively. Total alkaloid in ethyl acetate extract was counted as (10.46 ± 0.28)%(w/w), while lunacrine determined by ultra fast liquid chromatography-photodiode array detection method was found to be (3.55 ± 0.26)%(w/w). Conclusions: The high total alkaloid and lunacrine concentration on the extract confirm the potential cytotoxic property of ethyl acetate wood extract of L. amara.

Cytotoxic activity and phytochemical standardization of Lunasia amara Blanco wood extract

Objective To evaluate the cytotoxic activity of wood extracts of Lunasia amara Blanco (L. amara) and to perform further phytochemical standardization. Methods The wood extracts of L. amara were assessed for cytotoxic activity by in vitro tetrazolium bromide (MTT) method against two human cancer cell lines, cervical cancer cells (HeLa) and breast cancer cells (T47D). Thin layer chromatography, Dragendorf, acetic anhydride-sulfuric acid and ferric chloride were used to detect alkaloids, steroids and polyphenols, respectively. Furthermore, quantitative determination of total alkaloid by ultra fast liquid chromatography-photodiode array detection using lunacrine as a marker compound was performed as well. Results The ethyl acetate extract exhibited higher cell-growth inhibition than methanol and n-hexane extracts on HeLa and T47D cancer line cells with the IC

Research on anti T47 D breast cancer activity and its G protein-coupled estrogen receptor pathway of tanshinone IIA / 中国药理学通报

Aim To explore the effects of tanshinone IIA on cell proliferation via G protein-coupled estrogen receptor inductive and regulative pathway in typical es-trogen receptor and G protein-coupled estrogen receptor positive T47D breast cancer cells. Methods The pro-liferation rate of T47 D cells influenced by tanshinone IIA was analyzed by MTT assay. G protein-coupled es-trogen receptor agonist G1 and GPER antagonist G15 were employed as tools. GPER SiRNA was applied to build GPER gene silence T47D cells. GPER expres-sion influenced by tanshinone IIA was measured by Western blot. Results The proliferation rates of T47D cells treated with 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol· L-1 of tanshinone IIA were decreased significantly. Such effects could be attenuated by G1 or enhanced by G15 . Growth of GPER SiRNA transfected T47 D cells were significantly inhibited by 1 × 10 -5 mol·L-1 - 1 × 10 -7 mol·L-1 of tanshinone IIA treating. Result of Western blot showed that tanshinone IIA at 1 × 10 -5 mol· L-1 and 1 × 10 -6 mol · L-1 could induce de-crease of GPER protein expression in T47D cells. Conclusions Tanshinone IIA shows inhibitory effects on proliferation rate of T47 D breast cancer cells via GPER pathway. Tanshinone IIA could perform regula-tive function on GPER expression level in target cells.

Effect ofβ-sitosterol on T47D Cell Proliferation and Cell Cycle and Its Mechanisms / 世界科学技术-中医药现代化

This study was aimed to observe the influence of β-sitosterol (BSS) on estrogen receptor (ER) positive the human breast cancer cell line T47D and to study its mechanisms. ER antagonist ICI182 780 was employed to observe the influence on the proliferation. Proliferations of T47D cells influenced by different concentrations of BSS were analyzed by MTT assay. Cell cycle analyses were examined by flow cytometry. The protein expression of cyclin D1 was measured by western blot analysis and cyclin D1 mRNA was quantified by real-time PCR assay. The results showed that BSS in high dose exhibited significant inhibitory effects that were partly antagonized by ICI182 780 and decreased the proliferative index on T47D cells. However, BSS in low dose obviously promoted the proliferation that was completely inhibited by ICI182 780 and increased the proliferative index on T47D cells. The mRNA and protein levels of cyclin D1 were increased in low-dose BSS. The effect was blocked by ICI182 780. It was concluded that BSS in low concentration had phytoestrogenic effect by up-regulating the expression of cyclin D1 via ER pathway.

Cytotoxicity study of ethanol extract of the stem bark of asam kandis (Garcinia cowa Roxb.) on T47D breast cancer cell line

Objective: To investigate the cytotoxic effect of ethanol extract of the stem bark of asam kandis [Garcinia cowa Roxb. (G. cowa)] on T47D breast cancer cell line. Methods: The cytotoxicity of ethanol extract was carried out against human breast cancer cell line (T47D) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. The extract was added at various concentrations (0.1, 1, 10 and 100 μg/mL). The level of cytotoxicity was determined by calculating the level of IC

The effects of Hydnophytum formicarum ethanolic extract towards lymphocyte, vero and T47d cells proliferation in vitro.

This research aims were to explore the effects of Hydnophytum formicarum Jack (Rubiaceae) known as the Ant plant on proliferations of mouse lymphocyte, Vero and T47D's cell lines. Plant hypocotyls were collected from West Papua. Some parts were dried to gain plant with ant residue (A), while other parts were separated from the ant residue to gain plant without ant residue (B) and ant residue (C). After being macerated in ethanol and evaporated to dryness, each extract was tested on Balb/c mice lymphocytes and Vero cell lines by MTT assay in the presence and absence of Doxorubicin (Dox). Effect on T47D was observed by using Dox 25 g/mL as control. Data obtained were analyzed by ANOVA and paired sample T-test. Thin layer chromatography profiles were evaluated to identify the difference in chemical contents. The extracts A and B had the ability to increase lymphocyte proliferation by increasing concentration. Both extracts could not significantly enhanced the proliferation of Vero cells, neither in the presence nor absence of Dox. The extract B in 100 g/mL showed the highest inhibition against T47D cells by 53.45%. There was no chemical content difference observed on extracts A and B.

Cytotoxic activity and apoptosis induction of 8-hydroxyisocapnolactone-2´,3´-diol and its combination with Doxorubicin on MCF-7 and T47D cells.

The cytotoxic activity and apoptosis induction of 8-hydroxyisocapnolactone-2΄,3΄-diol (HICD) and its combination with doxorubicin (Doxo) on MCF-7 and T47D cells have been evaluated. The cytotoxic assay was performed using MTT method. The IC50 was used to express the cytotoxic potency, while the combination index (CI) was calculated to determine the effect of combination. The apoptotic assay was carried out using acrydine orange - ethidium bromide double staining method, while Bcl-2 and Bax expression were investigated by immunocytochemistry. The HICD exhibited cytotoxic activity on MCF-7 and T47D cells with the IC50 of 8 μg/ml (21.2 μM) and 4 μg/ml (10.6 μM), respectively. The combination of HICD with Doxo showed synergistic effect and increased apoptosis induction on both cell lines. The HICD did not affect Bcl-2 but increased Bax expression on MCF-7 cells, while on T47D cells it suppressed Bcl-2 expression but did not modulate Bax expression.

Cytotoxicity Studies of Xanthorrhizol and Its Mechanism Using Molecular Docking Simulation and Pharmacophore Modelling.

Xanthorrhizol (XNT) is one of major compounds from temulawak`s rhizome and its activity in several cancer cells is known. The aim of this study was to identify mechanism of xanthorrhizol from temulawak`s rhizome as an hERα inhibitor against breast cancer human cell lines. The cytotoxicity of XNT from temulawak`s rhizome on T47D human breast cancer cells lined by sulforhodamine B (SRB) method has been carried out, while molecular docking simulation and pharmacophore modelling methods were employed to predict mechanism of xanthorrhizol as hER inhibitor. Cytotoxicity studies showed that XNT of the isolated and standard had an IC50 100 and 55.50 μg/mL in T47D cells, respectively. Subsequently, molecular docking interaction showed that XNT might be able to compete with estradiol (E2) as a potential ERα inhibitor with the calculated binding free-energy of -8.2 kcal/mol, even the compound superimposed with tamoxifen (4-OHT). XNT formed hydrogen bonds with Arg394 and Glu353 as mention E2 and tamoxifen also formed same interaction with same residue and interacted hydrophobic bonds similar to 4-OHT with: Leu387, Leu384, Leu391, Phe404, L349, Leu346, Met388, and Leu525 of estrogen alpha Ligan Binding Domain (LBD), although 4-OHT indicated stronger hydrophobic when the tail of tamoxifen interacted with Tyr347, Asp351, Trp383 and Leu428. XNT missed two chemical features into HipHop models pharmacophore thus may result in reduced inhibitory activity against T47D compared than 4-OHT. The xanthorrhizol mechanism as a hER inhibitor is postulated as partial estrogen antagonist, is justifiable based on its competitive characteristic versus tamoxifen (OHT-200) which was located on the active side of HER-α.

Combinational effects of hexane insoluble fraction of Ficus septica Burm. F. and doxorubicin chemotherapy on T47D breast cancer cells

<p><b>OBJECTIVE</b>To evaluate the effects of n-hexane insoluble fraction (HIF) of Ficus septica leaves in combination with doxorubicin on cytotoxicity, cell cycle and apoptosis induction of breast cancer T47D cell lines.</p><p><b>METHODS</b>The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay. Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange. The expression of cleaved-poly (ADP-ribose) polymerase (PARP) on T47D cell lines was identified using immunocytochemistry.</p><p><b>RESULTS</b>The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells. In addition, combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis. HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase. The combination also exhibited upregulation of cleaved-PARP in T47D cells.</p><p><b>CONCLUSIONS</b>Based on this results, HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest. However, the molecular mechanism need to be explored further.</p>

Combination of low-concentration of novel phytoestrogen (8,9)-furanyl-pterocarpan-3-ol from Pachyrhizus erosus attenuated tamoxifen-associated growth inhibition on breast cancer T47D cells

Objective: To investigate the estrogenic effect of (8,9)-furanyl-pterocarpan-3-ol (FPC) on growth of human breast cancer T47D cells and the interactions between the FPC and tamoxifen (TAM), on the growth of estrogen receptor-dependent breast cancer T47D cells.Methods:T47D cells were treated with FPC alone (0.01-200 μmol/L) or in combination with TAM 20 nmol/L. The proliferation effect of FPC were conducted on T47D cells in vitro by MTT test. Furthermore, the expression of ERα or c-Myc were also determined by immunohistochemistry.Results:inhibitory effect on T47D cells, wheraes co-administered with low concentration (less than 1μmol/L) of FPC attenuated to promote cell proliferation. In contrast, the combination of TAM with higher doses (more than 20 μmol/L) of FPC showed growth inhibitory. This result was supported by immunocytochemistry studies that the administration of 20 nmol/L TAM down-regulated ER-αand c-Myc, but the combination of 20 nmol/L TAM and 1 μmol/L FPC robustly up-regulated expression of ER-α. Thus, the reduced growth inhibition of TAM 20 nmol/L by FPC 1 μmol/L on T47D cells may act via the modulation of ER-α.Conclusions:The findings indicate and suggest that FPC had estrogenic activity at low The results indicated that administration of an anti-estrogen TAM showed growth concentrations and anti-estrogenic effect that are likely to be regulated by c-Myc and estrogen receptors. We also confirm that low concentration of FPC attenuated the growth-inhibitory effects of TAM on mammary tumor prevention. Therefore, the present study suggests that caution is warranted regarding the consumption of dietary FPC by breast cancer patients while on TMA therapy.

Combinational effects of hexane insoluble fraction of Ficus septica Burm. F. and doxorubicin chemotherapy on T47D breast cancer cells

Objective: To evaluate the effects of n-hexane insoluble fraction (HIF) of Ficus septica leaves in combination with doxorubicin on cytotoxicity, cell cycle and apoptosis induction of breast cancer T47D cell lines. Methods: The in vitro drugs-stimulated cytotoxic effects were determined using MTT assay. Analysis of cell cycle distribution was performed using flowcytometer and the data was analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange. The expression of cleaved-poly (ADP-ribose) polymerase (PARP) on T47D cell lines was identified using immunocytochemistry. Results:The combination exhibited higher inhibitory effect on cell growth than the single treatment of doxorubicin in T47D cells. In addition, combination of doxorubicin and HIF increased the incidence of cells undergoing apoptosis. HIF could improve doxorubicin cytotoxic effect by changing the accumulation of cell cycle phase from G2/M to G1 phase. The combination also exhibited upregulation of cleaved-PARP in T47D cells. Conclusions: Based on this results, HIF is potential to be developed as co-chemotherapeutic agent for breast cancer by inducing apoptosis and cell cycle arrest. However, the molecular mechanism need to be explored further.

Effects of icariin and icaritin on proliferation of breast cancer T47D cells / 中草药

Objective: To investigate the effects of icariin (Ica) and icaritin (Ict) on the proliferationin of breast cancer T47D cells in vitro and to explore the relationship between estrogen receptor (ER) and the mechanisms. Methods: The ER positive T47D cells were treated with Ica and Ict at different concentration. The proliferation of T47D cells was measured by MTT method, and the cell cycle changes during the proliferation were further evaluated using flow cytometric assay. The effects of Ica and Ict on the expression of ERα and ERβ in T47D cells were analyzed by Western blotting. Results: Ica and Ict (10-9-10-6 mol/L) stimulated the proliferation of T47D cells, and the proliferation was inhibited by the estrogen antagonist ICI 182.780 (10-6 mol/L). Compared with the control group, Ica and Ict (10-7 mol/L) could increase the amount of T47D cells arrested in S phase, and the stimulation was inhibited by ICI 182.780. As well Ica and Ict (10-7 mol/L) could up-regulate the expression of ERα and ERβ proteins (P < 0.01). Conclusion: Ica and Ict stimulate the proliferation of T47D cells, and the effect may be performed through up-regulating the expression of ERα and ERβ proteins.

Research on the phytoestrogenic effect and its mechanism of isopsoralen in T47D cells / 中国药理学通报

Aim To study the phytoestrogenic effect and its mechanism of isopsoralen in estrogen receptor (ER) positive T47D and ER negative MDA-MB231 breast cancer cell line.Methods The proliferation rate and cell cycle distribution of T47D and MDA-MB231 cells influenced by isopsoralen were analyzed by MTT and flow cytometry assay respectively.PS2 mRNA level in T47D cells was quantified by Real-time RT-PCR assay.PS2 expression was measured by flow cytometry.Estrogen receptor antagonist ICI180,782 was employed as a tool.Results The proliferation rates and indexes of T47D cells treated with 1?10-6 mol?L-1 and 1?10-7 mol?L-1 isopsoralen were increased.These effects could be blocked by ICI182,780.Meanwhile,isopsoralen didn't show stimulative effects on proliferation rate of MDA-MB231 cells.Real-time RT-PCR and flow cytometry results showed that isopsoralen at 1?10-6 mol?L-1 and 1?10-7 mol?L-1 could induce elevation of pS2 mRNA and protein expression level in T47D cells.These effects could be partly blocked by ICI182,780.Conclusion Isopsoralen has phytoestrogenic effect and its effect is attained mainly via ER pathway.