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
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
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
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.
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.
