Objective:To determine the destructive ability of oxocrebanine,an anti-breast cancer active compound obtained from Stephania hainanensis H.S.Lo et Y.Tsoong,on microtubule network,and investigate the effect of oxocreba...Objective:To determine the destructive ability of oxocrebanine,an anti-breast cancer active compound obtained from Stephania hainanensis H.S.Lo et Y.Tsoong,on microtubule network,and investigate the effect of oxocrebanine on microtubule network homeostasis at both molecular and cellular levels.Methods:the EBI site competition method and molecular docking method were used to determine the occupation of the microtubule site of oxocrebanine.Western Blot was used to detect the effect of oxocrebanine on microtubule-associated proteins including STAT3,PAK1,CAMK4,and PKA.Results:The results of EBI site competition assay showed that the binding of EBI toβ-Tubulin covalent fusions produced adducts that appeared in regions of lower molecular weight thanβ-tubulin(ctrl 2).Molecular docking results showed that oxocrebanine could occupy the colchicine site of microtubule proteins.As revealed by Western Blot,the expression of STAT3 protein was decreased after MCF-7 cells have been treated with low,medium,and high concentration of oxocrebanine or the positive drug taxol for 48 h(P<0.01).The expression levels of PAK1 and Camk4 proteins aslo showed significant reductions(P<0.05,or P<0.01).Oxocrebanine also decreased the PKA protein in MCF-7 cells compared to the control group(P<0.01).Conclusions:Oxocrebanine,a ligand that binds at the colchicine site of tubulin,perturbs tubulin polymerization and causes mitosis in MCF-7 cells,thus leading to MCF-7 cell death.Oxocrebanine may promote microtubule dynamics through stathmin by inhibiting the expression levels of STAT3,PAK1,Camk4,and PKA proteins in MCF-7 cells.Oxocrebanine interfers with spindle formation,and ultimately causes mitotic catastrophe in MCF-7 cells.展开更多
Fucoidan is an active component of seaweed, which inhibits proliferation and induces apoptosis of several tumor cells while the detailed mechanisms underlying this process are still not clear. In this study, the effec...Fucoidan is an active component of seaweed, which inhibits proliferation and induces apoptosis of several tumor cells while the detailed mechanisms underlying this process are still not clear. In this study, the effect of Fucoidan on the proliferation and apoptosis of human breast cancer MCF-7 cells and the molecular mechanism of Fucoidan action were investigated. Viable cell number of MCF-7 cells was decreased by Fucoidan treatment in a dose-dependent manner as measured by MTT assay. Fucoidan treatment resulted in G1 phase arrest of MCF-7 cells as revealed by flow cytometry, which was associated with the decrease in the gene expression of cyclin D 1 and CDK-4. Annexin V/PI staining results showed that the number of apoptotic cells was associated with regulation of cytochrome C, cas- pase-8, Bax and Bcl-2 at transcriptional and translational levels. Both morphologic observation and Hoechst 33258 assay results confirmed the pro-apoptotic effect of Fucoidan. Meanwhile, the ROS pro- duction was also increased by Fucoidan treatment, which suggested that Fucoidan induced oxidative damage in MCF-7 cells. The results of present study demonstrated that Fucoidan could induce GI phase arrest and apoptosis in MCF-7 cells through regulating the cell cycle and apoptosis-related genes or proteins expression, and ROS generation is also involved in these processes.展开更多
Objective: To investigate whether dietary daidzein interact with endogenous 17β-Estradiol (E2) to give rise to additive or inhibitory effects on proliferation and apoptosis in breast cancer cells. Methods: Cell ...Objective: To investigate whether dietary daidzein interact with endogenous 17β-Estradiol (E2) to give rise to additive or inhibitory effects on proliferation and apoptosis in breast cancer cells. Methods: Cell cycle distribution and apoptosis induction were analyzed by using flow cytometry when breast cancer cell lines MCF-7 were cotreated with daidzein (1, 5 μmol/L) and E2 (0.1-10 nmol/L) for 5 days. Whether daidzein could alter E2-modulated mRNA expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERI3) and ERβ-estrogen response element (ERE) dependent transcription was investigated by RT-PCR and luciferase induction assays. The effects of daidzein on E2-modulated expression of proapoptotic p53, bax and antiapoptotic bcl-2 at both mRNA and protein levels were also investigated by RT-PCR and Western blot. Results: Daidzein enhanced the antiapoptotic effect in an Ea dose-dependent manner, but had no effect on E2-induced proliferation. Daidzein antagonized E2-induced ERβ mRNA expression and ERβ-ERE dependent transcription. In addition, daidzein only antagonized E2-upregulated expression of p53 and bax, but had no effect on E2-upregulated expression of bcl-2. Conclusion: Daidzein enhances the antiapoptotic effect of E2 on breast cancer cells by inhibiting E2-mediated p53-bax proapoptotic pathway. These results suggest that dietary daidzein may enhance deleterious effect of endogenous E2 in hormone-dependent breast cancer.展开更多
<span style="font-family:Verdana;">To synthesize, characterize and evaluate the antitumor potential derived from ruthenium compounds was generated in this study, from the precursor K[RuCl</span>&...<span style="font-family:Verdana;">To synthesize, characterize and evaluate the antitumor potential derived from ruthenium compounds was generated in this study, from the precursor K[RuCl</span><sub><span style="font-family:Verdana;">4</span></sub><span style="font-family:Verdana;">(bipy)] a route in a simple and reproducible synthesis for a novel compound of coordinating Ru</span><sup><span style="font-family:Verdana;">+3</span></sup><span style="font-family:Verdana;"> with bipy and L-trip. The spectroscopic characterization in the mi</span><span style="font-family:Verdana;">ddle infrared region (FTIR) shows the interactions between Ru-(L-trip), evidenced by the displacement of the carboxylate ion band for</span><span><span style="font-family:Verdana;"> higher energies, and also by the displacements of aliphatic amine bands, suggesting that bidentate coordination of the L-trip ligand occurred. Analysis of the results obtained with thermoanalytical techniques showed that the minimum formula of the compound, [RuCl</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">(bipy)(L-trip)]1/2H</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">O. Evaluation of the</span></span><span><span style="font-family:Verdana;"> antitumor potential of precursor K[RuCl</span><sub><span style="font-family:Verdana;">4</span></sub><span style="font-family:Verdana;">(bipy)] showed the toxic effects on MCF-7 cell line, but </span></span><span style="font-family:Verdana;">did not show selectivity and not reached PBMC cells to the same extent. The evaluation of the antitumor potential of the newly synthesized compound, [RuCl</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">(bipy)(L-trip)], demonstrated that the insertion of an L-tryptophan molecule into the precursor coordination sphere made it selective when compared to PBMC cells, for MCF-7 type tumor cells.</span>展开更多
Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferatio...Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.展开更多
Objective: To examine three plant extracts [Lumnitzera racemosa(Combretaceae)(L.racemosa), Albizia procera(Fabaceae)(A.procera) and Cananga odorata(Annonaceae)] for their potential as source of photosensitizers in pho...Objective: To examine three plant extracts [Lumnitzera racemosa(Combretaceae)(L.racemosa), Albizia procera(Fabaceae)(A.procera) and Cananga odorata(Annonaceae)] for their potential as source of photosensitizers in photodynamic therapy.Methods: Human mammary adenocarcinoma(MCF-7) cells were treated with the plant extracts, which were irradiated with 5.53 m W and 0.553 mW broadband light.Cell viability was assessed using MTT assay and induction of apoptosis was determined using terminal deoxynucleotidyl transferase-dUTP nick end labeling assay.Results: The crude ethanolic extracts, independently, were nontoxic against cancer and non-cancer cells but when irradiated with 5.53 mW broadband light, L.racemosa and A.procera extracts were cytotoxic against MCF-7 with IC_(50) of 11.63 mg/mL and10.73 mg/mL, respectively.With 0.553 mW broadband light, the IC_(50) values were higher at 17.14 mg/mL and 19.59 mg/mL, respectively.Photoactivated L.racemosa and A.procera extracts were found to be more cytotoxic against MCF-7 than the non-cancer cell line, human dermal fibroblast-neonatal.Moreover, the cytotoxicity of the extracts was mediated by apoptosis.Conclusions: Two of the plant extracts used, L.racemosa and A.procera were toxic and induced apoptosis to mammary cell adenocarcinoma, MCF-7 when photoactivated.These extracts were also more toxic to human cancer than non-cancer cell lines.展开更多
In my research,I studied the antioxidant effect of extracts of various rosemary samples obtained by Soxlet extraction on MCF-7 epithelial cells.I wanted to discover which of the extracts will have the highest content ...In my research,I studied the antioxidant effect of extracts of various rosemary samples obtained by Soxlet extraction on MCF-7 epithelial cells.I wanted to discover which of the extracts will have the highest content of total phenols determined in vitro with the Folin-Ciocalte reagent.The results show the highest content of total phenols in the rosemary sample from North Macedonia,so I assumed that it would have the biggest effective activity on MCF-7 cells.While doing the free radical inhibition test,I came to the conclusion that the plant with the highest antioxidant content does not necessarily have the best effect on the cells at different concentrations of the extracts.I suppose that it happens because antioxidants are polar and they need to transport through cell membrane conveyors whose number is limited in the membrane,that is,more antioxidants get assembled on the conveyors and cannot go through the membrane.展开更多
[Objectives] To investigate the mechanism of DNA damage of cisplatin( DDP),a broad spectrum anticancer drug on breast cancer MCF-7 cells,and to study the mechanism of apoptosis induced by DDP.[Methods]MCF-7 cells were...[Objectives] To investigate the mechanism of DNA damage of cisplatin( DDP),a broad spectrum anticancer drug on breast cancer MCF-7 cells,and to study the mechanism of apoptosis induced by DDP.[Methods]MCF-7 cells were treated by DDP( 0 mg/L,2 mg/L,4 mg/L,6 mg/L,6 mg/L,and 10 mg/L) for 48 hours. MTT assay was used to detect the inhibitory effect of DDP on MCF-7 cells and IC50 value was calculated. Western blot was adopted to detect the expression of γ-H2 AX,which was the marker of DNA double stranded breaks( DSBs) and ATM( sensory molecules of DSBs),the apoptotic signal transduction molecule cleaved caspase-3,and the proteins associated with apoptosis calpain.[Results]DDP inhibited MCF-7 cell activity in a concentration-dependent manner and IC50 was 7. 57 mg/L. In contrast to the control group( without DDP treatment),MCF-7 cells with DDP treatment expressed more γ-H2 AX,ATM,cleaved caspase-3 and calpain.[Conclusions] DDP could inhibit the activity of breast cancer MCF-7 cells. Its mechanisms may be associated with inhibition of MCF-7 cell apoptosis,induction of DNA double strand breaking and the expression of pro-apoptotic protein up-regulation.展开更多
The biochemical consequences induced by xenobiotic stress are featured in dose-response and timeresolved landscapes.Understanding the dynamic process of cellular adaptations is crucial in conducting the risk assessmen...The biochemical consequences induced by xenobiotic stress are featured in dose-response and timeresolved landscapes.Understanding the dynamic process of cellular adaptations is crucial in conducting the risk assessment for chemical exposure.As one of the most phenotype-related omics,metabolome in response to environmental stress can vary from seconds to days.Up to now,very few dynamic metabolomics studies have been conducted to provide time-dependent mechanistic interpretations in understanding xenobiotics-induced cellular adaptations.This study aims to explore the time-resolved metabolite dysregulation manner and dynamically perturbed biological functions in MCF-7 cells exposed to bisphenol A(BPA),a well-known endocrine-disrupting chemical.By sampling at 11 time points from several minutes to hours,thirty seven significantly dysregulated metabolites were identified,ranging from amino acids,fatty acids,carboxylic acids and nucleoside phosphate compounds.The metabolites in different pathways basically showed distinct time-resolved changing patterns,while those within the common class or same pathways showed similar and synchronized dysregulation behaviors.The pathway enrichment analysis suggested that purine metabolism,pyrimidine metabolism,aminoacyl-tRNA biosynthesis as well as glutamine/glutamate(GABA)metabolism pathways were heavily disturbed.As exposure event continued,MCF-7 cells went through multiple sequential metabolic adaptations from cell proliferation to energy metabolism,which indicated an enhancing cellular requirement for elevated energy homeostasis,oxidative stress response and ER-αmediated cell growth.We further focused on the time-dependent metabolite dysregulation behavior in purine and pyrimidine metabolism,and identified the impaired glycolysis and oxidative phosphorylation by redox imbalance.Lastly,we established a restricted cubic splinebased model to fit and predict metabolite’s full range dysregulation cartography,with metabolite’sensitivity comparisons retrieved and novel biomarkers suggested.Overall,the results indicated that 8 h BPA exposure leaded to global dynamic metabolome adaptions including amino acid,nucleoside and sugar metabolism disorders,and the dysregulated metabolites with interfered pathways at different stages are of significant temporal distinctions.展开更多
Activator protein-1(AP-1)transcription factor is a key component of many signal transduction pathways involved in the regulation of cellular processes and controls rapid responses of mammalian cells when exposed to th...Activator protein-1(AP-1)transcription factor is a key component of many signal transduction pathways involved in the regulation of cellular processes and controls rapid responses of mammalian cells when exposed to the variety of stimulus.The phorbol 12-myristate 13-acetate and Forskolin(Fo)are well-known kinase activators/stimulators of Protein Kinase C(PKC)and Protein Kinase A(PKA)respectively.Importantly,these kinases are found to be present in transitional points of many cell signaling pathways,especially those involved in proliferation.The stimulating effect of PKC and PKA on the expression of AP-1 factors in MCF-7 breast cell proliferation is not well characterized.Hence,the role of PKC by PMA treatment and the role of PKA by using Fo in MCF-7 cells is investigated.Where,cells treated with PMA showed increased cell proliferation,while Fo had no effect,but inhibited the PMA induced proliferation.The RT-PCR results showed the PMA induced c-Jun,c-Fos and Fra-1 expressions compared to control and Fo.However,Fo in combination with PMA,inhibit the PMA induced above mRNA expressions where Fo alone has no effect.Western blot studies validated the c-Jun expressions in PMA treated MCF-7 cells.Further,PMA increases the mRNA expression of Cyclin-E1,Cyclin-D1,and CDK-4,whereas Fo decreases their expressions.Thus,mitogenic effect of PMA and inhibitory action of Fo on MCF-7 cells is probably enhanced via activation of AP-1 factors and concomitant action of cell cycle regulators in the downstream singling cascade.展开更多
Cancer is one of the leading causes of death both in developing countries and across the globe.In Indonesia,cancer ranks as the fifth primary cause of death following heart disease,stroke,respiratory tract and diarrhe...Cancer is one of the leading causes of death both in developing countries and across the globe.In Indonesia,cancer ranks as the fifth primary cause of death following heart disease,stroke,respiratory tract and diarrhea.Therefore,studies on thiourea derivative compounds as anticancer agents have been profoundly conducted but still require further continuous development.In the present study,we aimed to synthesize new anticancer compounds of N-(phenylcarbamothioyl)-benzamide derivatives,namely N-(phenylcarbamothioyl)-4-bromobenzamide and N-(phenylcarbamothioyl)-4-fluorobenzamide compounds and assess their activities against MCF-7 breast cancer cells.The initial step was to predict the drug-receptor activity through docking between the tested compounds using epidermal growth factor receptor(EGFR)(PDB code: 1 M17).The compounds were futher synthesized from the reactions between benzoyl chloride derivatives and N-phenylthiourea.The structures of the new compounds were identified using FTIR,~1 H NMR,13 C NMR and mass spectra.The cytotoxic activities(IC_(50)) to breast cancer cells of MCF-7 N-(phenylcarbamothioyl)-4-bromobenzamide compound and N-(phenylcarbamothioyl)-4-fluorobenzamide were 0.27 mM and 0.31 mM,respectively.These two new compounds had better cytotoxic activities than those of the current hydroxyurea-based anticancer drugs(the reference compound) with an IC_(50) value of 9.76 mM.Furthermore,these two new compounds were not toxic to Vero normal cells.Therefore,they possessed tremendous potentials as the candidates for new drugs against breast cancer.展开更多
Objective:Realgar is a traditional mineral Chinese medicine with antitumor effects,but it has high toxicity and low efficacy in its crude form.The purpose of this study was to optimize realgar to increase its efficacy...Objective:Realgar is a traditional mineral Chinese medicine with antitumor effects,but it has high toxicity and low efficacy in its crude form.The purpose of this study was to optimize realgar to increase its efficacy and therapeutic potential.Methods:Crude realgar(CR)was mechanically ground to obtain nano-realgar(NR),and then nano-realgar processed products(NRPPs)were obtained using three different traditional Chinese medicine processing methods:grinding in water,acid water,and alkali water,respectively.Results:By analyzing the size distribution of nanoparticles and the content of arsenic trioxide(As_(2)O_(3);ATO),we found that acid water-ground NRPPs had the characteristics of high purity and low toxicity.The effects of CR,NR,and NRPPs on proliferation,cell cycle,and apoptosis of MCF-7 cells were detected,and the ability of NRPPs to induce apoptosis in MCF-7 cells was analyzed.The results showed that the average particle size of acid water-ground NRPPs was 137.7 nm,and the content of ATO was 2.83 mg/g.Acid water-ground NRPPs showed better effects on inhibiting proliferation,cell cycle,and apoptosis of MCF-7 cells than CR and NR.Western blot assays further confirmed that acid water-ground NRPPs upregulated the protein expression of TP53,Bax,cytochrome c,caspase-9,and caspase-3 in MCF-7 cells(P<0.05)and inhibited the expression of Bcl-2(P<0.05).Conclusion:These results suggest that acid water-ground NRPPs can induce apoptosis of MCF-7 cells through regulating mitochondrial-mediated apoptosis,providing evidence for the clinical application of realgar.展开更多
基金Natural Science Foundation of Hainan Province(No.820RC776)。
文摘Objective:To determine the destructive ability of oxocrebanine,an anti-breast cancer active compound obtained from Stephania hainanensis H.S.Lo et Y.Tsoong,on microtubule network,and investigate the effect of oxocrebanine on microtubule network homeostasis at both molecular and cellular levels.Methods:the EBI site competition method and molecular docking method were used to determine the occupation of the microtubule site of oxocrebanine.Western Blot was used to detect the effect of oxocrebanine on microtubule-associated proteins including STAT3,PAK1,CAMK4,and PKA.Results:The results of EBI site competition assay showed that the binding of EBI toβ-Tubulin covalent fusions produced adducts that appeared in regions of lower molecular weight thanβ-tubulin(ctrl 2).Molecular docking results showed that oxocrebanine could occupy the colchicine site of microtubule proteins.As revealed by Western Blot,the expression of STAT3 protein was decreased after MCF-7 cells have been treated with low,medium,and high concentration of oxocrebanine or the positive drug taxol for 48 h(P<0.01).The expression levels of PAK1 and Camk4 proteins aslo showed significant reductions(P<0.05,or P<0.01).Oxocrebanine also decreased the PKA protein in MCF-7 cells compared to the control group(P<0.01).Conclusions:Oxocrebanine,a ligand that binds at the colchicine site of tubulin,perturbs tubulin polymerization and causes mitosis in MCF-7 cells,thus leading to MCF-7 cell death.Oxocrebanine may promote microtubule dynamics through stathmin by inhibiting the expression levels of STAT3,PAK1,Camk4,and PKA proteins in MCF-7 cells.Oxocrebanine interfers with spindle formation,and ultimately causes mitotic catastrophe in MCF-7 cells.
基金supported by grants from The National Maritime Bureau Public Science and Technology Research Funds Projects of Ocean(No.201005013)the Wuhan Municipal Science and Technology Research Project of China(No.201260523185)
文摘Fucoidan is an active component of seaweed, which inhibits proliferation and induces apoptosis of several tumor cells while the detailed mechanisms underlying this process are still not clear. In this study, the effect of Fucoidan on the proliferation and apoptosis of human breast cancer MCF-7 cells and the molecular mechanism of Fucoidan action were investigated. Viable cell number of MCF-7 cells was decreased by Fucoidan treatment in a dose-dependent manner as measured by MTT assay. Fucoidan treatment resulted in G1 phase arrest of MCF-7 cells as revealed by flow cytometry, which was associated with the decrease in the gene expression of cyclin D 1 and CDK-4. Annexin V/PI staining results showed that the number of apoptotic cells was associated with regulation of cytochrome C, cas- pase-8, Bax and Bcl-2 at transcriptional and translational levels. Both morphologic observation and Hoechst 33258 assay results confirmed the pro-apoptotic effect of Fucoidan. Meanwhile, the ROS pro- duction was also increased by Fucoidan treatment, which suggested that Fucoidan induced oxidative damage in MCF-7 cells. The results of present study demonstrated that Fucoidan could induce GI phase arrest and apoptosis in MCF-7 cells through regulating the cell cycle and apoptosis-related genes or proteins expression, and ROS generation is also involved in these processes.
基金supported by the National Natural Science Foundation of China (No.30671508)by State Key Laboratory for Agrobiotechnology of China (No.2009SKLAB07-5)
文摘Objective: To investigate whether dietary daidzein interact with endogenous 17β-Estradiol (E2) to give rise to additive or inhibitory effects on proliferation and apoptosis in breast cancer cells. Methods: Cell cycle distribution and apoptosis induction were analyzed by using flow cytometry when breast cancer cell lines MCF-7 were cotreated with daidzein (1, 5 μmol/L) and E2 (0.1-10 nmol/L) for 5 days. Whether daidzein could alter E2-modulated mRNA expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERI3) and ERβ-estrogen response element (ERE) dependent transcription was investigated by RT-PCR and luciferase induction assays. The effects of daidzein on E2-modulated expression of proapoptotic p53, bax and antiapoptotic bcl-2 at both mRNA and protein levels were also investigated by RT-PCR and Western blot. Results: Daidzein enhanced the antiapoptotic effect in an Ea dose-dependent manner, but had no effect on E2-induced proliferation. Daidzein antagonized E2-induced ERβ mRNA expression and ERβ-ERE dependent transcription. In addition, daidzein only antagonized E2-upregulated expression of p53 and bax, but had no effect on E2-upregulated expression of bcl-2. Conclusion: Daidzein enhances the antiapoptotic effect of E2 on breast cancer cells by inhibiting E2-mediated p53-bax proapoptotic pathway. These results suggest that dietary daidzein may enhance deleterious effect of endogenous E2 in hormone-dependent breast cancer.
文摘<span style="font-family:Verdana;">To synthesize, characterize and evaluate the antitumor potential derived from ruthenium compounds was generated in this study, from the precursor K[RuCl</span><sub><span style="font-family:Verdana;">4</span></sub><span style="font-family:Verdana;">(bipy)] a route in a simple and reproducible synthesis for a novel compound of coordinating Ru</span><sup><span style="font-family:Verdana;">+3</span></sup><span style="font-family:Verdana;"> with bipy and L-trip. The spectroscopic characterization in the mi</span><span style="font-family:Verdana;">ddle infrared region (FTIR) shows the interactions between Ru-(L-trip), evidenced by the displacement of the carboxylate ion band for</span><span><span style="font-family:Verdana;"> higher energies, and also by the displacements of aliphatic amine bands, suggesting that bidentate coordination of the L-trip ligand occurred. Analysis of the results obtained with thermoanalytical techniques showed that the minimum formula of the compound, [RuCl</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">(bipy)(L-trip)]1/2H</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">O. Evaluation of the</span></span><span><span style="font-family:Verdana;"> antitumor potential of precursor K[RuCl</span><sub><span style="font-family:Verdana;">4</span></sub><span style="font-family:Verdana;">(bipy)] showed the toxic effects on MCF-7 cell line, but </span></span><span style="font-family:Verdana;">did not show selectivity and not reached PBMC cells to the same extent. The evaluation of the antitumor potential of the newly synthesized compound, [RuCl</span><sub><span style="font-family:Verdana;">2</span></sub><span style="font-family:Verdana;">(bipy)(L-trip)], demonstrated that the insertion of an L-tryptophan molecule into the precursor coordination sphere made it selective when compared to PBMC cells, for MCF-7 type tumor cells.</span>
基金This work was supported by NationalNatural Science Foundation of China (No. 30200095).
文摘Objective: Our previous studies have firstly demonstrated that 17β -E2 up-regulates LRP16 gene expression in human breast cancer MCF-7 cells, and ectopic expression of the LRP16 gene promotes MCF-7 cells proliferation. Here, the effects of the LRP16 gene expression on growth of MCF-7 human breast cancer cells and the mechanism were further studied by establishing two stably LRP16-inhibitory MCR-7 cell lines. Methods: Hairpin small interference RNA (siRNA) strategy, by which hairpin siRNA was released by U6 promoter and was mediated by pLPC-based retroviral vector, was adopted to knockdown endogenous LRP16 level in MCF-7 cells. And the hairpin siRNA against green fluorescence protein (GFP) was used as the negative control. The suppressant efficiency of the LRP16 gene expression was confirmed by Nothern blot. Cell proliferation assay and soft agar colony formation assay were used to determine the status of the cells proliferation. Cell cycle checkpoints including cyclin E and cyclin D1 were examined by Western blot. Results: The results from cell proliferation assays suggested that down-regulation of LRP16 gene expression is capable of inhibiting MCF-7 breast cancer cell growth and down-regulation of the LRP16 gene expression is able to inhibit anchorage-independent growth of breast cancer cells in soft agar. We also demonstrated that cyclin E and cyclin D1 proteins were much lower in the LRP16-inhibitory cells than in the control cells. Conclusion: These data suggest that LRP16 gene play an important role in MCF-7 cells proliferation by regulating the pathway of the G1/S transition and may function as an important modulator in regulating the process of tumorigenesis in human breast.
基金Supported by Institute of Biology,University of the Philippines,Diliman through TA#9774-362-499-439
文摘Objective: To examine three plant extracts [Lumnitzera racemosa(Combretaceae)(L.racemosa), Albizia procera(Fabaceae)(A.procera) and Cananga odorata(Annonaceae)] for their potential as source of photosensitizers in photodynamic therapy.Methods: Human mammary adenocarcinoma(MCF-7) cells were treated with the plant extracts, which were irradiated with 5.53 m W and 0.553 mW broadband light.Cell viability was assessed using MTT assay and induction of apoptosis was determined using terminal deoxynucleotidyl transferase-dUTP nick end labeling assay.Results: The crude ethanolic extracts, independently, were nontoxic against cancer and non-cancer cells but when irradiated with 5.53 mW broadband light, L.racemosa and A.procera extracts were cytotoxic against MCF-7 with IC_(50) of 11.63 mg/mL and10.73 mg/mL, respectively.With 0.553 mW broadband light, the IC_(50) values were higher at 17.14 mg/mL and 19.59 mg/mL, respectively.Photoactivated L.racemosa and A.procera extracts were found to be more cytotoxic against MCF-7 than the non-cancer cell line, human dermal fibroblast-neonatal.Moreover, the cytotoxicity of the extracts was mediated by apoptosis.Conclusions: Two of the plant extracts used, L.racemosa and A.procera were toxic and induced apoptosis to mammary cell adenocarcinoma, MCF-7 when photoactivated.These extracts were also more toxic to human cancer than non-cancer cell lines.
文摘In my research,I studied the antioxidant effect of extracts of various rosemary samples obtained by Soxlet extraction on MCF-7 epithelial cells.I wanted to discover which of the extracts will have the highest content of total phenols determined in vitro with the Folin-Ciocalte reagent.The results show the highest content of total phenols in the rosemary sample from North Macedonia,so I assumed that it would have the biggest effective activity on MCF-7 cells.While doing the free radical inhibition test,I came to the conclusion that the plant with the highest antioxidant content does not necessarily have the best effect on the cells at different concentrations of the extracts.I suppose that it happens because antioxidants are polar and they need to transport through cell membrane conveyors whose number is limited in the membrane,that is,more antioxidants get assembled on the conveyors and cannot go through the membrane.
基金Supported by Project of National Natural Science Foundation(30600753&81172154)Hunan Provincial Natural Science Foundation of China(2016JJ2088)+1 种基金Project of Hunan Provincial Department of Education(13C541)Project of Hunan Administration of Traditional Chinese Medicine(2015123)
文摘[Objectives] To investigate the mechanism of DNA damage of cisplatin( DDP),a broad spectrum anticancer drug on breast cancer MCF-7 cells,and to study the mechanism of apoptosis induced by DDP.[Methods]MCF-7 cells were treated by DDP( 0 mg/L,2 mg/L,4 mg/L,6 mg/L,6 mg/L,and 10 mg/L) for 48 hours. MTT assay was used to detect the inhibitory effect of DDP on MCF-7 cells and IC50 value was calculated. Western blot was adopted to detect the expression of γ-H2 AX,which was the marker of DNA double stranded breaks( DSBs) and ATM( sensory molecules of DSBs),the apoptotic signal transduction molecule cleaved caspase-3,and the proteins associated with apoptosis calpain.[Results]DDP inhibited MCF-7 cell activity in a concentration-dependent manner and IC50 was 7. 57 mg/L. In contrast to the control group( without DDP treatment),MCF-7 cells with DDP treatment expressed more γ-H2 AX,ATM,cleaved caspase-3 and calpain.[Conclusions] DDP could inhibit the activity of breast cancer MCF-7 cells. Its mechanisms may be associated with inhibition of MCF-7 cell apoptosis,induction of DNA double strand breaking and the expression of pro-apoptotic protein up-regulation.
基金supported by Singapore Ministry of Education Academic Research Fund Tier 1(No.04MNP000567C120)Startup Grant of Fudan University(No.JIH 1829010Y).
文摘The biochemical consequences induced by xenobiotic stress are featured in dose-response and timeresolved landscapes.Understanding the dynamic process of cellular adaptations is crucial in conducting the risk assessment for chemical exposure.As one of the most phenotype-related omics,metabolome in response to environmental stress can vary from seconds to days.Up to now,very few dynamic metabolomics studies have been conducted to provide time-dependent mechanistic interpretations in understanding xenobiotics-induced cellular adaptations.This study aims to explore the time-resolved metabolite dysregulation manner and dynamically perturbed biological functions in MCF-7 cells exposed to bisphenol A(BPA),a well-known endocrine-disrupting chemical.By sampling at 11 time points from several minutes to hours,thirty seven significantly dysregulated metabolites were identified,ranging from amino acids,fatty acids,carboxylic acids and nucleoside phosphate compounds.The metabolites in different pathways basically showed distinct time-resolved changing patterns,while those within the common class or same pathways showed similar and synchronized dysregulation behaviors.The pathway enrichment analysis suggested that purine metabolism,pyrimidine metabolism,aminoacyl-tRNA biosynthesis as well as glutamine/glutamate(GABA)metabolism pathways were heavily disturbed.As exposure event continued,MCF-7 cells went through multiple sequential metabolic adaptations from cell proliferation to energy metabolism,which indicated an enhancing cellular requirement for elevated energy homeostasis,oxidative stress response and ER-αmediated cell growth.We further focused on the time-dependent metabolite dysregulation behavior in purine and pyrimidine metabolism,and identified the impaired glycolysis and oxidative phosphorylation by redox imbalance.Lastly,we established a restricted cubic splinebased model to fit and predict metabolite’s full range dysregulation cartography,with metabolite’sensitivity comparisons retrieved and novel biomarkers suggested.Overall,the results indicated that 8 h BPA exposure leaded to global dynamic metabolome adaptions including amino acid,nucleoside and sugar metabolism disorders,and the dysregulated metabolites with interfered pathways at different stages are of significant temporal distinctions.
基金Grants Commission(UGC),New Delhi for providing Major Research Project financial Grant(F.No.34-250/2008(SR))to SCS.
文摘Activator protein-1(AP-1)transcription factor is a key component of many signal transduction pathways involved in the regulation of cellular processes and controls rapid responses of mammalian cells when exposed to the variety of stimulus.The phorbol 12-myristate 13-acetate and Forskolin(Fo)are well-known kinase activators/stimulators of Protein Kinase C(PKC)and Protein Kinase A(PKA)respectively.Importantly,these kinases are found to be present in transitional points of many cell signaling pathways,especially those involved in proliferation.The stimulating effect of PKC and PKA on the expression of AP-1 factors in MCF-7 breast cell proliferation is not well characterized.Hence,the role of PKC by PMA treatment and the role of PKA by using Fo in MCF-7 cells is investigated.Where,cells treated with PMA showed increased cell proliferation,while Fo had no effect,but inhibited the PMA induced proliferation.The RT-PCR results showed the PMA induced c-Jun,c-Fos and Fra-1 expressions compared to control and Fo.However,Fo in combination with PMA,inhibit the PMA induced above mRNA expressions where Fo alone has no effect.Western blot studies validated the c-Jun expressions in PMA treated MCF-7 cells.Further,PMA increases the mRNA expression of Cyclin-E1,Cyclin-D1,and CDK-4,whereas Fo decreases their expressions.Thus,mitogenic effect of PMA and inhibitory action of Fo on MCF-7 cells is probably enhanced via activation of AP-1 factors and concomitant action of cell cycle regulators in the downstream singling cascade.
基金Directorate General of Resources for Science,Technology and Higher Education of Ministry of Research,Technology and Higher Education(KEMRISTEK DIKTI)with scheme of scholarship funding for PhD program at University of Airlangga,Surabaya,Indonesia
文摘Cancer is one of the leading causes of death both in developing countries and across the globe.In Indonesia,cancer ranks as the fifth primary cause of death following heart disease,stroke,respiratory tract and diarrhea.Therefore,studies on thiourea derivative compounds as anticancer agents have been profoundly conducted but still require further continuous development.In the present study,we aimed to synthesize new anticancer compounds of N-(phenylcarbamothioyl)-benzamide derivatives,namely N-(phenylcarbamothioyl)-4-bromobenzamide and N-(phenylcarbamothioyl)-4-fluorobenzamide compounds and assess their activities against MCF-7 breast cancer cells.The initial step was to predict the drug-receptor activity through docking between the tested compounds using epidermal growth factor receptor(EGFR)(PDB code: 1 M17).The compounds were futher synthesized from the reactions between benzoyl chloride derivatives and N-phenylthiourea.The structures of the new compounds were identified using FTIR,~1 H NMR,13 C NMR and mass spectra.The cytotoxic activities(IC_(50)) to breast cancer cells of MCF-7 N-(phenylcarbamothioyl)-4-bromobenzamide compound and N-(phenylcarbamothioyl)-4-fluorobenzamide were 0.27 mM and 0.31 mM,respectively.These two new compounds had better cytotoxic activities than those of the current hydroxyurea-based anticancer drugs(the reference compound) with an IC_(50) value of 9.76 mM.Furthermore,these two new compounds were not toxic to Vero normal cells.Therefore,they possessed tremendous potentials as the candidates for new drugs against breast cancer.
基金supported by the Science and Technology_Research Project of Hubei Education Department(No.B2019097).
文摘Objective:Realgar is a traditional mineral Chinese medicine with antitumor effects,but it has high toxicity and low efficacy in its crude form.The purpose of this study was to optimize realgar to increase its efficacy and therapeutic potential.Methods:Crude realgar(CR)was mechanically ground to obtain nano-realgar(NR),and then nano-realgar processed products(NRPPs)were obtained using three different traditional Chinese medicine processing methods:grinding in water,acid water,and alkali water,respectively.Results:By analyzing the size distribution of nanoparticles and the content of arsenic trioxide(As_(2)O_(3);ATO),we found that acid water-ground NRPPs had the characteristics of high purity and low toxicity.The effects of CR,NR,and NRPPs on proliferation,cell cycle,and apoptosis of MCF-7 cells were detected,and the ability of NRPPs to induce apoptosis in MCF-7 cells was analyzed.The results showed that the average particle size of acid water-ground NRPPs was 137.7 nm,and the content of ATO was 2.83 mg/g.Acid water-ground NRPPs showed better effects on inhibiting proliferation,cell cycle,and apoptosis of MCF-7 cells than CR and NR.Western blot assays further confirmed that acid water-ground NRPPs upregulated the protein expression of TP53,Bax,cytochrome c,caspase-9,and caspase-3 in MCF-7 cells(P<0.05)and inhibited the expression of Bcl-2(P<0.05).Conclusion:These results suggest that acid water-ground NRPPs can induce apoptosis of MCF-7 cells through regulating mitochondrial-mediated apoptosis,providing evidence for the clinical application of realgar.