Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ab...Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ability,the arginine-glycine-aspartic(RGD)peptide and the epidermal growth factor receptor interference(EGFRi)peptide were fused with MAP30,which was named ELRL-MAP30.The efficiency of targeted therapy for triple-negative breast cancer(TNBC)MDA-MB-231 cells,which lack the expression of estrogen receptor(ER),Progesterone receptor(PgR)and human epidermal growth factor receptor-2(HER2),is limited.In this study,we focus on exploring the effect and mechanism of ELRL-MAP30 on TNBC MDA-MB-231 cells.First,we discovered that ELRL-MAP30 significantly inhibited the migration and invasion of MDA-MB-231 cells and induced MDA-MB-231 cell apoptosis.Moreover,ELRL-MAP30 treatment resulted in a significant increase in Bax expression and a decrease in Bcl-2 expression.Furthermore,ELRL-MAP30 triggered apoptosis via the Fak/EGFR/Erk and Ilk/Akt signaling pathways.In addition,recombinant ELRL-MAP30 can inhibit chicken embryonic angiogenesis,and also inhibit the tube formation ability of human umbilical vein endothelial cells(HUVECs),indicating its potential therapeutic effects on tumor angiogenesis.Collectively,these results indicate that ELRL-MAP30 has significant tumor-targeting properties in MDA-MB-231 cancer cells and reveals potential therapeutic effects on angiogenesis.These findings indicate the potential role of ELRL-MAP30 in the targeted treatment of the TNBC cell line MDA-MB-231.展开更多
Objective:To investigate the mechanism of anticancer activity of a pigment OR3 from Streptomyces coelicolor in in vitro and in vivo metastatic breast cancer models and to characterize the pigment.Methods:The anticance...Objective:To investigate the mechanism of anticancer activity of a pigment OR3 from Streptomyces coelicolor in in vitro and in vivo metastatic breast cancer models and to characterize the pigment.Methods:The anticancer mechanism was analyzed in MDA-MB-231 cells using MTT,lactate dehydrogenase,caspase,DNA fragmentation,clonogenic,flow cytometry,Western blot,and scratch assays.The effects of OR3 on xenograft mouse models were evaluated by tumor volume measurement,hematological analysis,and histopathological observation.The characterization of OR3 was also performed using gas chromatograohy-mass spectrometry and nuclear magnetic resonance spectroscopy.Results:OR3 exhibited potent cytotoxicity against MDA-MB-231 cells,with no observed effects on HEK-293 cells.Caspase-9 activation was detected in OR3-treated MDA-MB-231 cells.Flow cytometry showed a dose-dependent induction of apoptosis and cell cycle arrest at the sub-G_(1) and S phases.Furthermore,OR3 completely inhibited MDA-MB-231 cell migration and demonstrated anti-proliferative effects by downregulating the protein expression of KPNA2,XPO1,RAB5B,and p38 MAPK.In in vivo studies,OR3 was non-toxic to mice,inhibited tumor xenograft growth,and maintained normal hematological parameters and tissue architecture.Nuclear magnetic resonance spectroscopy demonstrated the presence of a prodigiosin-like compound,while gas chromatography-mass spectrometry analysis identified additional compounds in OR3.Conclusions:Our findings validate OR3 pigment as a promising compound for the treatment of metastatic breast cancer,warranting further studies.展开更多
Gambogic acid （GA） is an anticancer agent in phase IIb clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins ...Gambogic acid （GA） is an anticancer agent in phase IIb clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins of GA in cancer cells using proteomic method and establish possible network using bioinformatic analysis. Cytotoxicity and anti-migration effects of GA in MDA-MB-231 cells were checked using MTT assay, flow cytometry, wound migration assay, and chamber migration assay. Possible target-related proteins of GA at early （3 h） and late stage （24 h） of treatment were searched using a proteomic technology, two-dimensional electro- phoresis （2-DE）. The possible network of GA was established using bioinformatic analysis. The intracellular expression levels of vimentin, keratin 18, and calumenin were determined using Western blotting. GA inhibited cell proliferation and induced cell cycle arrest at G2/M phase and apoptosis in MDA-MB-231 cells. Additionally, GA exhibited anti-migration effects at non-toxic doses. In 2-DE analysis, totally 23 possible GA targeted proteins were found, including those with functions in cytoskeleton and transport, regulation of redox state, metabolism, ubiquitin-proteasome system, transcription and translation, protein transport and modification, and cytokine. Network analysis of these proteins suggested that cytoskeleton-related proteins might play important roles in the effects of GA. Results of Western blotting confirmed the cleavage of vimentin, increase in keratin 18, and decrease in calumenin levels in GA-treated cells. In summary, GA is a multi-target compound and its anti-cancer effects may be based on several target-related pro- teins such as cytoskeleton-related proteins.展开更多
OBJECTIVE： To test the synergistic effects of the aqueous extract of Tubeimu （Rhizoma Bolbosternmatis） and Fuzi （Radix Aconiti Lateralis Preparata） on MDA-MB-231 and SKBR3 breast cancer cells.METHODS： A combined...OBJECTIVE： To test the synergistic effects of the aqueous extract of Tubeimu （Rhizoma Bolbosternmatis） and Fuzi （Radix Aconiti Lateralis Preparata） on MDA-MB-231 and SKBR3 breast cancer cells.METHODS： A combined index was created for the effects of Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts. Cell proliferation was performed by trypan blue exclusion and 3-（4,5-dimethylthiazol-2-yl）-5-（3- carboxy- methoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium （MTS） assays. Flow cytometry was used to assess cell cycle distribution and apoptosis. Cell migration was determined by wound-healing and transwell assays. Confocal microscopy was used to detect E-cadherin and actin filaments. RESULTS： The aqueous extract from Tubeimu （Rhizoma Bolbostemmatis）and Fuzi （Radix Aconiti lateralis Preparata） exerted synergetic effects on the growth of MDA-MB-231 cells and G1 phase arrest. When exposed to extracts at concentrations of 62.5 ：62.5 and 62.5：31.3 μg/mL, the combination index was 0.83 and 0.74, respectively. Interestingly, 62.5： 31.3 μg/mL of combined drugs enhanced the inhibitory effect of Tubeimu （Rhizoma Bolbostemmatis） on the migration of SKBR3 cells and reduced the stimulative effect of Fuzi （Radix Aconiti Lateralis Preparata） （P 〈 0.01）, in which cells showed an increased expression of E-cadherin and reorganization of actin filaments （P 〈 0.001）. 62.5：62.5 μg/mL extract also synergistically induced apoptosis of MDA-MB-231 cells （P 〈 0.05 and P 〈 0.001）. Acting as the main active ingredients in the extract, tubeimoside I and acetylbenzoylaconine at 10：10μg/ mL and 5：2.5 μg/mL also produced inhibitory effects on the proliferation and migration of cells （P 〈 0.01）. CONCLUSION： Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts had synergic effects on MDA-MB-231 and SKBR3 cells.展开更多
文摘Momordica antiviral protein 30 kD(MAP30)is a type I ribosome-inactivating protein(RIP)with antibacterial,anti-HIV and antitumor activities but lacks the ability to target tumor cells.To increase its tumor-targeting ability,the arginine-glycine-aspartic(RGD)peptide and the epidermal growth factor receptor interference(EGFRi)peptide were fused with MAP30,which was named ELRL-MAP30.The efficiency of targeted therapy for triple-negative breast cancer(TNBC)MDA-MB-231 cells,which lack the expression of estrogen receptor(ER),Progesterone receptor(PgR)and human epidermal growth factor receptor-2(HER2),is limited.In this study,we focus on exploring the effect and mechanism of ELRL-MAP30 on TNBC MDA-MB-231 cells.First,we discovered that ELRL-MAP30 significantly inhibited the migration and invasion of MDA-MB-231 cells and induced MDA-MB-231 cell apoptosis.Moreover,ELRL-MAP30 treatment resulted in a significant increase in Bax expression and a decrease in Bcl-2 expression.Furthermore,ELRL-MAP30 triggered apoptosis via the Fak/EGFR/Erk and Ilk/Akt signaling pathways.In addition,recombinant ELRL-MAP30 can inhibit chicken embryonic angiogenesis,and also inhibit the tube formation ability of human umbilical vein endothelial cells(HUVECs),indicating its potential therapeutic effects on tumor angiogenesis.Collectively,these results indicate that ELRL-MAP30 has significant tumor-targeting properties in MDA-MB-231 cancer cells and reveals potential therapeutic effects on angiogenesis.These findings indicate the potential role of ELRL-MAP30 in the targeted treatment of the TNBC cell line MDA-MB-231.
文摘Objective:To investigate the mechanism of anticancer activity of a pigment OR3 from Streptomyces coelicolor in in vitro and in vivo metastatic breast cancer models and to characterize the pigment.Methods:The anticancer mechanism was analyzed in MDA-MB-231 cells using MTT,lactate dehydrogenase,caspase,DNA fragmentation,clonogenic,flow cytometry,Western blot,and scratch assays.The effects of OR3 on xenograft mouse models were evaluated by tumor volume measurement,hematological analysis,and histopathological observation.The characterization of OR3 was also performed using gas chromatograohy-mass spectrometry and nuclear magnetic resonance spectroscopy.Results:OR3 exhibited potent cytotoxicity against MDA-MB-231 cells,with no observed effects on HEK-293 cells.Caspase-9 activation was detected in OR3-treated MDA-MB-231 cells.Flow cytometry showed a dose-dependent induction of apoptosis and cell cycle arrest at the sub-G_(1) and S phases.Furthermore,OR3 completely inhibited MDA-MB-231 cell migration and demonstrated anti-proliferative effects by downregulating the protein expression of KPNA2,XPO1,RAB5B,and p38 MAPK.In in vivo studies,OR3 was non-toxic to mice,inhibited tumor xenograft growth,and maintained normal hematological parameters and tissue architecture.Nuclear magnetic resonance spectroscopy demonstrated the presence of a prodigiosin-like compound,while gas chromatography-mass spectrometry analysis identified additional compounds in OR3.Conclusions:Our findings validate OR3 pigment as a promising compound for the treatment of metastatic breast cancer,warranting further studies.
基金supported by the Twelfth Five-Year National Science & Technology Support Program(No.2012BAI 29B06)Shanghai Science & Technology Support Program(No.13431900 401)+4 种基金China Postdoctoral Science Foundation Funded Project(No.2012M5 10907)Shanghai Postdoctoral Scientific Program(No.13R21417800)the Postdoctor Research Program of Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences(No.2012KIP516)the Sanofi-Aventis-Shanghai Institutes for Biological Sciences Scholarship Programthe National Nature Science Foundation(Nos.81302809 and 81373964)
文摘Gambogic acid （GA） is an anticancer agent in phase IIb clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins of GA in cancer cells using proteomic method and establish possible network using bioinformatic analysis. Cytotoxicity and anti-migration effects of GA in MDA-MB-231 cells were checked using MTT assay, flow cytometry, wound migration assay, and chamber migration assay. Possible target-related proteins of GA at early （3 h） and late stage （24 h） of treatment were searched using a proteomic technology, two-dimensional electro- phoresis （2-DE）. The possible network of GA was established using bioinformatic analysis. The intracellular expression levels of vimentin, keratin 18, and calumenin were determined using Western blotting. GA inhibited cell proliferation and induced cell cycle arrest at G2/M phase and apoptosis in MDA-MB-231 cells. Additionally, GA exhibited anti-migration effects at non-toxic doses. In 2-DE analysis, totally 23 possible GA targeted proteins were found, including those with functions in cytoskeleton and transport, regulation of redox state, metabolism, ubiquitin-proteasome system, transcription and translation, protein transport and modification, and cytokine. Network analysis of these proteins suggested that cytoskeleton-related proteins might play important roles in the effects of GA. Results of Western blotting confirmed the cleavage of vimentin, increase in keratin 18, and decrease in calumenin levels in GA-treated cells. In summary, GA is a multi-target compound and its anti-cancer effects may be based on several target-related pro- teins such as cytoskeleton-related proteins.
基金Supported by Funding Program of(National Science Foundation of China(Mechanism of Warming and Relieving Cold Phlegm Therapy Inhibiting Tumor Metastasis from the Field of Biomechanical,No.81503383)Science and Technology Development of Tianjin Municipal Higher Education(Compatibility Principle of Warming and Relieving Cold Phlegm Formula for the Treatment of Human Hepatocellular Carcinoma,No.20110105)Tianjin city application basis and cutting-edge technology research program(Effect of Herbs Formula Radix Aconiti Praeparata-Rhizoma Bolbostemmatis on Breast Cancer,No.13JCQNJC13800)
文摘OBJECTIVE： To test the synergistic effects of the aqueous extract of Tubeimu （Rhizoma Bolbosternmatis） and Fuzi （Radix Aconiti Lateralis Preparata） on MDA-MB-231 and SKBR3 breast cancer cells.METHODS： A combined index was created for the effects of Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts. Cell proliferation was performed by trypan blue exclusion and 3-（4,5-dimethylthiazol-2-yl）-5-（3- carboxy- methoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium （MTS） assays. Flow cytometry was used to assess cell cycle distribution and apoptosis. Cell migration was determined by wound-healing and transwell assays. Confocal microscopy was used to detect E-cadherin and actin filaments. RESULTS： The aqueous extract from Tubeimu （Rhizoma Bolbostemmatis）and Fuzi （Radix Aconiti lateralis Preparata） exerted synergetic effects on the growth of MDA-MB-231 cells and G1 phase arrest. When exposed to extracts at concentrations of 62.5 ：62.5 and 62.5：31.3 μg/mL, the combination index was 0.83 and 0.74, respectively. Interestingly, 62.5： 31.3 μg/mL of combined drugs enhanced the inhibitory effect of Tubeimu （Rhizoma Bolbostemmatis） on the migration of SKBR3 cells and reduced the stimulative effect of Fuzi （Radix Aconiti Lateralis Preparata） （P 〈 0.01）, in which cells showed an increased expression of E-cadherin and reorganization of actin filaments （P 〈 0.001）. 62.5：62.5 μg/mL extract also synergistically induced apoptosis of MDA-MB-231 cells （P 〈 0.05 and P 〈 0.001）. Acting as the main active ingredients in the extract, tubeimoside I and acetylbenzoylaconine at 10：10μg/ mL and 5：2.5 μg/mL also produced inhibitory effects on the proliferation and migration of cells （P 〈 0.01）. CONCLUSION： Tubeimu （Rhizoma Bolbostemmatis） and Fuzi （Radix Aconiti Lateralis Preparata） extracts had synergic effects on MDA-MB-231 and SKBR3 cells.
