3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic c...3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic conditions.Our previous studies demonstrated that YC-1 inhibited breast cancer cell proliferation under normoxic conditions.In the current study,we investigated the targets of YC-1 and mechanism of its action in MDA-MB-468 breast cancer cells.In the in vitro experiments,we found that YC-1 significantly inhibited MDA-MB-468 cell proliferation in normoxia and hypoxia.Under normoxic conditions,YC-1 induced apoptosis of MDA-MB-468 cells and blocked cell cycle in the G1 phase,and these effects were possibly related to caspase 8,p21,and p27 expression.RT-PCR and Western blotting results showed that YC-1 primarily inhibited HIF-1α at the mRNA and protein levels under hypoxic conditions,but suppressed the expression of epidermal growth factor receptor(EGFR) at the mRNA and protein levels under normoxic conditions.In vivo,YC-1 prolonged survival,increased survival rate,decreased tumor size and metastasis rate,and inhibited tissue EGFR and HIF-1α expression.However,YC-1 exerted no obvious effect on body weight.These results indicate that YC-1 inhibits the proliferation of MDA-MB-468 cells by acting on multiple targets with minimal side effects.Thus,YC-1 is a promising target drug for breast cancer.展开更多
目的:探讨miR-28-3p在三阴性乳腺癌(triple negative breast cancer,TNBC)组织和细胞系中的表达及其对MDA-MB-468细胞恶性生物学行为的影响。方法:收集2013年1月至2014年1月河北医科大学第四医院乳腺中心手术切除的、经病理证实的83例女...目的:探讨miR-28-3p在三阴性乳腺癌(triple negative breast cancer,TNBC)组织和细胞系中的表达及其对MDA-MB-468细胞恶性生物学行为的影响。方法:收集2013年1月至2014年1月河北医科大学第四医院乳腺中心手术切除的、经病理证实的83例女性TNBC患者的癌组织和癌旁组织标本,以及TNBC细胞系MDA-MB-468、HCC-1937、MDA-MB-231、MDA-MB-436、MDA-MB-453和人正常乳腺上皮细胞MCF10A,用qPCR检测组织和细胞系中miR-28-3p的表达水平并分析其表达与患者临床病理特征的相关性。用miR-28-3p抑制剂转染MDA-MB-468细胞后,用CCK-8、流式细胞术、细胞划痕和Transwell实验分别检测miR-28-3p抑制剂对MDA-MB-468细胞增殖、凋亡、侵袭和迁移能力的影响,用Western blotting检测MDA-MB-468细胞中桥接整合因子1(bridging integrator-1,BIN1)蛋白的表达水平。通过生物信息学工具预测miR-28-3p的靶基因BIN1,用双荧光素酶报告基因实验验证miR-28-3p对BIN1的调控作用。结果:TNBC组织及细胞系中miR-28-3p表达水平显著高于癌旁组织及MCF10A细胞(均P<0.01);83例TNBC组织中共有56例(67.47%)高表达miR-28-3p,其高表达与患者的Ki-67表达水平、肿瘤大小和TNM分期密切相关(均P<0.05或P<0.01)。与miR-NC组比较,miR-28-3p抑制剂组MDA-MB-468细胞增殖、侵袭和迁移能力降低,凋亡率升高(均P<0.05或P<0.01)。双荧光素酶报告基因和实验证实BIN1是miR-28-3p的靶基因,miR-28-3p抑制剂可上调MDA-MB-468细胞中BIN1蛋白的表达(P<0.05)。结论:miR-28-3p在TNBC组织及细胞中呈高表达状态,mi R-28-3p抑制剂上调BIN1表达进而抑制MDA-MB-468细胞的增殖、迁移和侵袭能力,并促进其凋亡。展开更多
Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate...Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate and methanol by Soxhlet extraction.Aqueous extract was prepared by decoction at 50 ℃.Extracts of A.precatorius leaves were used to treat selected cancer and normal cell lines for72 h.Furthermore,3-(4,5-dimethyl thiazol-2-yl)2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability.Analysis of cell cycle arrest,apoptosis assay and apoptosis protein expressions were determined by flow cytometry.Results:Methanolic extract of A.precatorius leaves showed the lowest IC50 on MDA-MB-231 cells at(26.40±5.40)μg/mL.Flow cytometry analysis revealed that cell arrest occurred at G0/G1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB-231 cells treated with methanolic extract of A.precatorius leaves.Methanolic extract of A.precatorius leaves induced apoptosis by upregulation of Bax,p53 and caspase-3 and downregulation of Bcl-2.Conclusions:Methanolic extract of A precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway.展开更多
Objective： To elucidate the effects of amlodipine on the proliferation and apoptosis of human breast carcinoma MDA-MB-231 cells. Methods： Light microscopy was used to determine the effects of amlodipine on cell morp...Objective： To elucidate the effects of amlodipine on the proliferation and apoptosis of human breast carcinoma MDA-MB-231 cells. Methods： Light microscopy was used to determine the effects of amlodipine on cell morphology; Flow cytometry was used to quantitate cells undergoing apoptosis; the expression of a cell cycle-related protein, proliferating cell nuclear antigen （PCNA） and an antiapoptosis protein, Bcl-2 were assessed by immunocytochemistry. Results： Amlodipine concentration of 8.25umol/L （1/2 of ICs0） affected the morphology, decreased the expression of PCNA and Bcl-2 and induced apoptosis of human breast carcinoma MDA-MB-231 cells. Conclusion： The effect of amlodipine on the antiproliferation of human breast carcinoma MDA-MB-231 cells is related to inducement of apoptosis, and the decrease of the expression of Bcl-2 and PCNA may be the possible mechanism for proliferation inhibitory and inducement of apoptosis.展开更多
OBJECTIVE To investigate the anti-proliferative effect of rosiglitazone and its relationship to peroxisome proliferator-activated receptor γ （PPARγ） in human breast cancer cell line MDA-MB-231 and evaluate the pot...OBJECTIVE To investigate the anti-proliferative effect of rosiglitazone and its relationship to peroxisome proliferator-activated receptor γ （PPARγ） in human breast cancer cell line MDA-MB-231 and evaluate the potential application value of rosiglitazone for breast cancer therapy. METHODS The cytostatic effect of rosiglitazone on MDA- MB-231 cells was measured by the MTT assay. Cell-cycle kinetics was assessed by flow cytometry. Apoptotic cells were determined by the TUNEL assay. MDA-MB-231 cells were treated with rosiglitazone or in combination with the PPARy antagonist GW9662 to investigate the effect of rosiglitazone on cell proliferation and its relationship to PPARγ. RESULTS The results showed that rosiglitazone could inhibit growth of MDA-MB-231 cells in a dose- and time-dependent manner with an IC50 value of 5.2 μmol/L at 24 h after the drug was added into the culture. Cell cycle analysis showed that the percentage of G0/G1 phase cells increased, S phase cells decreased, and cells were arrested in G1 phase with increasing concentrations of rosiglitazone. Detectable signs of apoptotic cell death caused by rosiglitazone occurred at a concentration of 100 μmol/L and the apoptotic rate was （18 ± 3）%. PPARγ selective antagonist GW9662 could partially reverse the inhibitory effect of rosiglitazone on proliferation of MDA-MB-231 cells. CONCLUSION It was concluded that rosiglitazone can inhibit growth of MDA-MB-231 cells via PPARy activation and a high concentration of rosiglitazone can also induce MDA-MB-231 cell apoptosis. These results suggest that PPARy represents a putative molecular target for chemopreventive therapy and rosiglitazone may be effective in the treatment of breast cancer.展开更多
To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly ...To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA-MD-468.After transfection,the cells were selected by G418.The resistant clones were chosen and expanded in DMEM culture medium.RT-PCR,immunohistochemical method and western blot were used to determine the expression of target genes.Results An anti-G418 cell clone was established and expanded in culture.The transfected PTEN gene MDA-MD-468 cells showed expression of PTEN mRNA and PTEN protein.Conclusion Human breast cancer cell line MDA-MB-468 established in this study expresses consistently exogenous PTEN genes.4 refs,6 figs.展开更多
Breast cancer,a predominant global health issue,requires ongoing exploration of new therapeutic strategies.Palbociclib(PAL),a well-known cyclin-dependent kinase(CDK)inhibitor,plays a critical role in breast cancer tre...Breast cancer,a predominant global health issue,requires ongoing exploration of new therapeutic strategies.Palbociclib(PAL),a well-known cyclin-dependent kinase(CDK)inhibitor,plays a critical role in breast cancer treatment.While its efficacy is recognized,the interplay between PAL and cellular autophagy,particularly in the context of the RAF/MEK/ERK signaling pathway,remains insufficiently explored.This study investigates PAL’s inhibitory effects on breast cancer using both in vitro(MCF7 and MDA-MB-468 cells)and in vivo(tumor-bearing nude mice)models.Aimed at elucidating the impact of PAL on autophagic processes and exploring the potential of combining it with trametinib(TRA),an MEK inhibitor,our research seeks to address the challenge of PAL-induced drug resistance.Ourfindings reveal that PAL significantly decreases the viability of MCF7 and MDA-MB-468 cells and reduces tumor size in mice while showing minimal cytotoxicity in MCF10A cells.However,PAL also induces protective autophagy,potentially leading to drug resistance via the RAF/MEK/ERK pathway activation.Introducing TRA effectively neutralized this autophagy,enhancing PAL’s anti-tumor efficacy.A combination of PAL and TRA synergistically reduced cell viability and proliferation,and in vivo studies showed notable tumor size reduction.In conclusion,the PAL and TRA combination emerges as a promising strategy for overcoming PAL-induced resistance,offering a new horizon in breast cancer treatment.展开更多
Objective:To investigate the effect of piperine on human breast cancer cells.Methods:The effect of piperine on proliferation and migration of human breast cancer cells,MCF-7 and MDA-MB-231,was investigated using colon...Objective:To investigate the effect of piperine on human breast cancer cells.Methods:The effect of piperine on proliferation and migration of human breast cancer cells,MCF-7 and MDA-MB-231,was investigated using colony formation assays,wound healing assays,Matrigel migration assays,flow cytometry,RT-qPCR,and Western blotting assays.Results:Piperine inhibited the growth of MCF-7 and MDA-MB-231 cells and suppressed colony formation.Cell reduction at the G_(0)/G_(1) phase and cell arrest at the G_(2)/M phase were observed in breast cancer cells.However,the significant effect was only demonstrated in MDA-MB-231 cells.Moreover,cancer cell migration was suppressed by piperine at low concentration.RT-qPCR and Western blotting assays showed that piperine downregulated Rac1 gene and protein expression.Conclusions:Piperine could inhibit growth and migration of breast cancer cells by reducing Rac1 gene and protein expression.展开更多
Objective： To investigate the effect of RNA interfering on VEGF-C in MDA-MB-231 cells. Methods： Three small interfering RNAs （siRNAa, siRNAb, siRNAc） were prepared. The most efficient one was screened and short ha...Objective： To investigate the effect of RNA interfering on VEGF-C in MDA-MB-231 cells. Methods： Three small interfering RNAs （siRNAa, siRNAb, siRNAc） were prepared. The most efficient one was screened and short hairpin （shRNA） was designed, the recombinant plasmid pGenesil-1/VEGF-C was constructed, and transfeeted into MDA-MB-231 cells by Lipofectamine TM 2000. RT-PCR, Western-blot an immunohistochemical methods were performed to detect the expression of VEGF-C. Results： RT-PCR results showed that siRNAa, siRNAb, siRNAc could inhibit the growth of MDA-MB-231 cells, among which, siRNAa was the most significant, with an inhibition rate of 72.1%. The recombinant plasmid pGenesil-1/VEGF-C was successfully constructed using shRNA and pGenesil-1. VEGF-C expression was significantly inhibited as determined by RT-PCR, immunocytochemistry staining and Western blot （P〈0.05）. Conclusion： shRNA RNAi technology could silence the expression of VEGF-C in MDA-MB-231 cells, which suggested that the technology may be one of the effective methods for inhibiting lymphangiogenesis in breast cancer.展开更多
Synthetic phosphoethanolamine(Pho-s)is a monophosphoester ester with anti-inflammatory and pro-apoptotic properties.Meclizine chloridrate(MC)is a histamine H1 receptor blocker that is also able to inhibit cellular res...Synthetic phosphoethanolamine(Pho-s)is a monophosphoester ester with anti-inflammatory and pro-apoptotic properties.Meclizine chloridrate(MC)is a histamine H1 receptor blocker that is also able to inhibit cellular respiration.However,MC does not inhibit cellular respiration in isolated mitochondria such as antimycin and rotenone.Methyl-β-cyclodextrin(MβCD)belongs to theβ-cyclodextrin family,which is capable of removing cholesterol from the plasma membrane.The aim of this study was to evaluate the proliferative effects of meclizine chloridrate and methyl-β-cyclodextrin compounds associated with synthetic phosphoethanolamine in a triple-negative human breast tumor line,MDA-MB-231 Cell viability of the tumor line and normal cells FN1 was evaluated by MTT colorimetric test;the production of free radicals was determined by lipoperoxidation(LPO)test;and the percentage of cell cycle phases and proliferative index was evaluated by flow cytometry.Cell viability demonstrated a significant decrease with the treatments of MβCD,MC and Pho-s associated with MC.The production of free radicals decreases significantly in all treatments.In addition,a significant increase of DNA fragment and decrease in G0/G1 cell cycle phase were observed in cellular percentage with concentrations of 20 and 30 mM of Pho-s in association with MC and MβCD,respectively.展开更多
The process of in situ tumors developing into malignant tumors and exhibiting invasive behavior is extremely complicated.From a biophysical point of view,it is a phase change process affected by many factors,including...The process of in situ tumors developing into malignant tumors and exhibiting invasive behavior is extremely complicated.From a biophysical point of view,it is a phase change process affected by many factors,including cell-to-cell,cell-to-chemical material,cell-to-environment interaction,etc.In this study,we constructed spheroids based on green fluorescence metastatic breast cancer cells MDA-MB-231 to simulate malignant tumors in vitro,while constructed a three-dimensional(3D)biochip to simulate a micro-environment for the growth and invasion of spheroids.In the experiment,the 3D spheroid was implanted into the chip,and the oriented collagen fibers controlled by collagen concentration and injection rate could guide the MDA-MB-231 cells in the spheroid to undergo directional invasion.The experiment showed that the oriented fibers greatly accelerated the invasion speed of MDA-MB-231 cells compared with the traditional uniform tumor micro-environment,namely obvious invasive branches appeared on the spheroids within 24 hours.In order to analyze this interesting phenomenon,we have developed a quantitative analyzing approach to explore strong angle correlation between the orientation of collagen fibers and invasive direction of cancer cell.The results showed that the oriented collagen fibers produced by the chip can greatly stimulate the invasion potential of cancer cells.This biochip is not only conducive to modeling cancer cell metastasis and studying cell invasion mechanisms,but also has the potential to build a quantitative evaluation platform that can be used in future chemical drug treatments.展开更多
基金supported by a grant from Jilin Province Science and Technology Development Project(No.200905198)
文摘3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole(YC-1),the hypoxia-inducible factor-1 alpha(HIF-1α) inhibitor,suppresses tumor proliferation and metastasis by down-regulating HIF-1α expression under hypoxic conditions.Our previous studies demonstrated that YC-1 inhibited breast cancer cell proliferation under normoxic conditions.In the current study,we investigated the targets of YC-1 and mechanism of its action in MDA-MB-468 breast cancer cells.In the in vitro experiments,we found that YC-1 significantly inhibited MDA-MB-468 cell proliferation in normoxia and hypoxia.Under normoxic conditions,YC-1 induced apoptosis of MDA-MB-468 cells and blocked cell cycle in the G1 phase,and these effects were possibly related to caspase 8,p21,and p27 expression.RT-PCR and Western blotting results showed that YC-1 primarily inhibited HIF-1α at the mRNA and protein levels under hypoxic conditions,but suppressed the expression of epidermal growth factor receptor(EGFR) at the mRNA and protein levels under normoxic conditions.In vivo,YC-1 prolonged survival,increased survival rate,decreased tumor size and metastasis rate,and inhibited tissue EGFR and HIF-1α expression.However,YC-1 exerted no obvious effect on body weight.These results indicate that YC-1 inhibits the proliferation of MDA-MB-468 cells by acting on multiple targets with minimal side effects.Thus,YC-1 is a promising target drug for breast cancer.
基金funded by the Universiti Sains Malaysia Short Term Grant(304/PPSP/61313046)
文摘Objective:To determine the anti-proliferative activity of Abrus precatorius(A.precatorius)leaf extracts and their effect on cell death.Methods:A.precatorius leaves were extracted successively with hexane,ethyl acetate and methanol by Soxhlet extraction.Aqueous extract was prepared by decoction at 50 ℃.Extracts of A.precatorius leaves were used to treat selected cancer and normal cell lines for72 h.Furthermore,3-(4,5-dimethyl thiazol-2-yl)2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability.Analysis of cell cycle arrest,apoptosis assay and apoptosis protein expressions were determined by flow cytometry.Results:Methanolic extract of A.precatorius leaves showed the lowest IC50 on MDA-MB-231 cells at(26.40±5.40)μg/mL.Flow cytometry analysis revealed that cell arrest occurred at G0/G1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB-231 cells treated with methanolic extract of A.precatorius leaves.Methanolic extract of A.precatorius leaves induced apoptosis by upregulation of Bax,p53 and caspase-3 and downregulation of Bcl-2.Conclusions:Methanolic extract of A precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway.
文摘Objective： To elucidate the effects of amlodipine on the proliferation and apoptosis of human breast carcinoma MDA-MB-231 cells. Methods： Light microscopy was used to determine the effects of amlodipine on cell morphology; Flow cytometry was used to quantitate cells undergoing apoptosis; the expression of a cell cycle-related protein, proliferating cell nuclear antigen （PCNA） and an antiapoptosis protein, Bcl-2 were assessed by immunocytochemistry. Results： Amlodipine concentration of 8.25umol/L （1/2 of ICs0） affected the morphology, decreased the expression of PCNA and Bcl-2 and induced apoptosis of human breast carcinoma MDA-MB-231 cells. Conclusion： The effect of amlodipine on the antiproliferation of human breast carcinoma MDA-MB-231 cells is related to inducement of apoptosis, and the decrease of the expression of Bcl-2 and PCNA may be the possible mechanism for proliferation inhibitory and inducement of apoptosis.
文摘OBJECTIVE To investigate the anti-proliferative effect of rosiglitazone and its relationship to peroxisome proliferator-activated receptor γ （PPARγ） in human breast cancer cell line MDA-MB-231 and evaluate the potential application value of rosiglitazone for breast cancer therapy. METHODS The cytostatic effect of rosiglitazone on MDA- MB-231 cells was measured by the MTT assay. Cell-cycle kinetics was assessed by flow cytometry. Apoptotic cells were determined by the TUNEL assay. MDA-MB-231 cells were treated with rosiglitazone or in combination with the PPARy antagonist GW9662 to investigate the effect of rosiglitazone on cell proliferation and its relationship to PPARγ. RESULTS The results showed that rosiglitazone could inhibit growth of MDA-MB-231 cells in a dose- and time-dependent manner with an IC50 value of 5.2 μmol/L at 24 h after the drug was added into the culture. Cell cycle analysis showed that the percentage of G0/G1 phase cells increased, S phase cells decreased, and cells were arrested in G1 phase with increasing concentrations of rosiglitazone. Detectable signs of apoptotic cell death caused by rosiglitazone occurred at a concentration of 100 μmol/L and the apoptotic rate was （18 ± 3）%. PPARγ selective antagonist GW9662 could partially reverse the inhibitory effect of rosiglitazone on proliferation of MDA-MB-231 cells. CONCLUSION It was concluded that rosiglitazone can inhibit growth of MDA-MB-231 cells via PPARy activation and a high concentration of rosiglitazone can also induce MDA-MB-231 cell apoptosis. These results suggest that PPARy represents a putative molecular target for chemopreventive therapy and rosiglitazone may be effective in the treatment of breast cancer.
文摘To investigate exogenous PTEN gene transfected human breast cancer cell line MDA-MD-468.Methods Using the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA-MD-468.After transfection,the cells were selected by G418.The resistant clones were chosen and expanded in DMEM culture medium.RT-PCR,immunohistochemical method and western blot were used to determine the expression of target genes.Results An anti-G418 cell clone was established and expanded in culture.The transfected PTEN gene MDA-MD-468 cells showed expression of PTEN mRNA and PTEN protein.Conclusion Human breast cancer cell line MDA-MB-468 established in this study expresses consistently exogenous PTEN genes.4 refs,6 figs.
基金supported by the Sichuan Science and Technology Program(Grant Nos.2020YJ0494,24GJHZ0058,21RCYJ0021,and 2022YFS0620)the National Natural Science Foundation of China(Grant No.81903829)+1 种基金the Southwest Medical University Science and Technology Program(Grant Nos.2021ZKZD015,2021ZKZD018,and 2021ZKMS046)the Macao Science and Technology Development Fund of Macao SAR(Project Nos.SKLQRCM(MUST)-2020-2022 and MUST-SKL-2021-005).
文摘Breast cancer,a predominant global health issue,requires ongoing exploration of new therapeutic strategies.Palbociclib(PAL),a well-known cyclin-dependent kinase(CDK)inhibitor,plays a critical role in breast cancer treatment.While its efficacy is recognized,the interplay between PAL and cellular autophagy,particularly in the context of the RAF/MEK/ERK signaling pathway,remains insufficiently explored.This study investigates PAL’s inhibitory effects on breast cancer using both in vitro(MCF7 and MDA-MB-468 cells)and in vivo(tumor-bearing nude mice)models.Aimed at elucidating the impact of PAL on autophagic processes and exploring the potential of combining it with trametinib(TRA),an MEK inhibitor,our research seeks to address the challenge of PAL-induced drug resistance.Ourfindings reveal that PAL significantly decreases the viability of MCF7 and MDA-MB-468 cells and reduces tumor size in mice while showing minimal cytotoxicity in MCF10A cells.However,PAL also induces protective autophagy,potentially leading to drug resistance via the RAF/MEK/ERK pathway activation.Introducing TRA effectively neutralized this autophagy,enhancing PAL’s anti-tumor efficacy.A combination of PAL and TRA synergistically reduced cell viability and proliferation,and in vivo studies showed notable tumor size reduction.In conclusion,the PAL and TRA combination emerges as a promising strategy for overcoming PAL-induced resistance,offering a new horizon in breast cancer treatment.
基金financially supported by Mahasarakham University 2021(MSU2021).
文摘Objective:To investigate the effect of piperine on human breast cancer cells.Methods:The effect of piperine on proliferation and migration of human breast cancer cells,MCF-7 and MDA-MB-231,was investigated using colony formation assays,wound healing assays,Matrigel migration assays,flow cytometry,RT-qPCR,and Western blotting assays.Results:Piperine inhibited the growth of MCF-7 and MDA-MB-231 cells and suppressed colony formation.Cell reduction at the G_(0)/G_(1) phase and cell arrest at the G_(2)/M phase were observed in breast cancer cells.However,the significant effect was only demonstrated in MDA-MB-231 cells.Moreover,cancer cell migration was suppressed by piperine at low concentration.RT-qPCR and Western blotting assays showed that piperine downregulated Rac1 gene and protein expression.Conclusions:Piperine could inhibit growth and migration of breast cancer cells by reducing Rac1 gene and protein expression.
基金supported by a grant from the Key Project of Chongqing Science and Technology Committee(No.2005AC0076)
文摘Objective： To investigate the effect of RNA interfering on VEGF-C in MDA-MB-231 cells. Methods： Three small interfering RNAs （siRNAa, siRNAb, siRNAc） were prepared. The most efficient one was screened and short hairpin （shRNA） was designed, the recombinant plasmid pGenesil-1/VEGF-C was constructed, and transfeeted into MDA-MB-231 cells by Lipofectamine TM 2000. RT-PCR, Western-blot an immunohistochemical methods were performed to detect the expression of VEGF-C. Results： RT-PCR results showed that siRNAa, siRNAb, siRNAc could inhibit the growth of MDA-MB-231 cells, among which, siRNAa was the most significant, with an inhibition rate of 72.1%. The recombinant plasmid pGenesil-1/VEGF-C was successfully constructed using shRNA and pGenesil-1. VEGF-C expression was significantly inhibited as determined by RT-PCR, immunocytochemistry staining and Western blot （P〈0.05）. Conclusion： shRNA RNAi technology could silence the expression of VEGF-C in MDA-MB-231 cells, which suggested that the technology may be one of the effective methods for inhibiting lymphangiogenesis in breast cancer.
文摘Synthetic phosphoethanolamine(Pho-s)is a monophosphoester ester with anti-inflammatory and pro-apoptotic properties.Meclizine chloridrate(MC)is a histamine H1 receptor blocker that is also able to inhibit cellular respiration.However,MC does not inhibit cellular respiration in isolated mitochondria such as antimycin and rotenone.Methyl-β-cyclodextrin(MβCD)belongs to theβ-cyclodextrin family,which is capable of removing cholesterol from the plasma membrane.The aim of this study was to evaluate the proliferative effects of meclizine chloridrate and methyl-β-cyclodextrin compounds associated with synthetic phosphoethanolamine in a triple-negative human breast tumor line,MDA-MB-231 Cell viability of the tumor line and normal cells FN1 was evaluated by MTT colorimetric test;the production of free radicals was determined by lipoperoxidation(LPO)test;and the percentage of cell cycle phases and proliferative index was evaluated by flow cytometry.Cell viability demonstrated a significant decrease with the treatments of MβCD,MC and Pho-s associated with MC.The production of free radicals decreases significantly in all treatments.In addition,a significant increase of DNA fragment and decrease in G0/G1 cell cycle phase were observed in cellular percentage with concentrations of 20 and 30 mM of Pho-s in association with MC and MβCD,respectively.
基金Project supported by the National Natural Science Foundation of China(Grant Nos.11974066 and 11674043)the Fundamental Research Funds for the Central Universities,China(Grant No.2019CDYGYB007)the Natural Science Foundation of Chongqing,China(Grant No.cstc2019jcyj-msxmX0477).
文摘The process of in situ tumors developing into malignant tumors and exhibiting invasive behavior is extremely complicated.From a biophysical point of view,it is a phase change process affected by many factors,including cell-to-cell,cell-to-chemical material,cell-to-environment interaction,etc.In this study,we constructed spheroids based on green fluorescence metastatic breast cancer cells MDA-MB-231 to simulate malignant tumors in vitro,while constructed a three-dimensional(3D)biochip to simulate a micro-environment for the growth and invasion of spheroids.In the experiment,the 3D spheroid was implanted into the chip,and the oriented collagen fibers controlled by collagen concentration and injection rate could guide the MDA-MB-231 cells in the spheroid to undergo directional invasion.The experiment showed that the oriented fibers greatly accelerated the invasion speed of MDA-MB-231 cells compared with the traditional uniform tumor micro-environment,namely obvious invasive branches appeared on the spheroids within 24 hours.In order to analyze this interesting phenomenon,we have developed a quantitative analyzing approach to explore strong angle correlation between the orientation of collagen fibers and invasive direction of cancer cell.The results showed that the oriented collagen fibers produced by the chip can greatly stimulate the invasion potential of cancer cells.This biochip is not only conducive to modeling cancer cell metastasis and studying cell invasion mechanisms,but also has the potential to build a quantitative evaluation platform that can be used in future chemical drug treatments.
