目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄...目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄糖调节蛋白78(glucose regulated protein of 78,GRP78)的表达情况;分析临床资料与mH2A和GRP78表达的关系;免疫共沉淀实验检测GRP78和mH2A蛋白的相互作用;Western印迹检测mH2A和GRP78的相关作用及Butein与mH2A之间的作用。Transwell试验检测mH2A的表达对黑色素瘤细胞侵袭能力的影响;划痕试验检测mH2A的表达对黑色素瘤细胞迁移能力的影响;Western印迹检测沉默mH2A后细胞外信号调节激酶1(ERK1)和MAPK/ERK激酶(MEK)蛋白的表达。结果:与癌旁正常组织相比,黑色素瘤组织中mH2A和GRP78表达明显降低(P<0.05);Butein可以增强mH2A的表达水平(P<0.05),mH2A可以调节GRP78蛋白的表达(P<0.05);沉默mH2A促进黑色素瘤A375细胞的侵袭迁移能力,上调MEK和ERK1蛋白的表达。结论:Butein可以增强mH2A靶向GRP78蛋白通过MAPK信号通路抑制黑色素瘤细胞的迁移和侵袭行为。展开更多
AIM: To evaluate the effects of butein on inflammatory cytokines, matrix metalloproteinase-9(MMP-9), andcolitis in interleukin(IL)-10-/- mice.METHODS: To synchronize colitis, 8- to 10-wk-old IL-10-/- mice were fed pel...AIM: To evaluate the effects of butein on inflammatory cytokines, matrix metalloproteinase-9(MMP-9), andcolitis in interleukin(IL)-10-/- mice.METHODS: To synchronize colitis, 8- to 10-wk-old IL-10-/- mice were fed pellet-chow containing piroxicam for 2 wk. Subsequently, phosphate-buffered saline or butein(1 mg/kg per day, ip) was injected for 4 wk. Histologic scores, inflammatory cytokines, MMP-9 and phosphorylated signal transducer and activator of transcription 3(p STAT3) expressions were analyzed in IL-10-/- mice and in Colo 205 cells.RESULTS: Butein reduced the colonic inflammatory score by > 50%. Expression levels of IL-6, IL-1β, interferon(IFN)-γ and MMP-9 were decreased in the colons of mice exposed to butein, whereas other inflammatory cytokines(IL-17 A, IL-21 and IL-22) were unchanged. Immunohistochemical staining for p STAT3 and MMP-9 was significantly decreased in the buteintreated groups compared with the controls. Butein inhibited IL-6-induced activation of STAT3 in Colo 205 cells.CONCLUSION: Butein ameliorated colitis in IL-10-/-mice by regulating IL-6/STAT3 and MMP-9 activation.展开更多
Aim:Malignant pleural mesothelioma is a chemoresistant tumor,and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma(MPM)patients.We obtained the microRNA expression pro...Aim:Malignant pleural mesothelioma is a chemoresistant tumor,and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma(MPM)patients.We obtained the microRNA expression profile of three biphasic-sarcomatoid MPM cell lines to identify commonly expressed microRNAs and evaluate the effect of butein,a chemo-sensitizing compound,on this microRNA subset.Methods:Nanostring-based microRNA profiling and analysis through the ROSALIND platform were employed to identify the commonly modulated microRNAs and their targets.MicroRNA-mimic transfection,Luciferase assay,and Western blotting were employed to show specific perturbation of TWIST1 levels by miR-186-5p.Sphere-forming assays,invasion assay,and metabolic profiling were used to assess the biological consequences of the butein-instigated miR-186-5p-mediated perturbation of TWIST1 levels.TGCA analysis was used to search for the correlation between TWIST1 and miR-186-5p levels in biphasic and epithelioid MPM specimens.Results:We identified a set of perturbed microRNAs,common to three biphasic/sarcomatoid MPM cell lines,after butein treatment.When focusing on miR-186-5p,we unraveled a butein-ignited and miR-186-5p-mediated modulation of TWIST1 levels which affected the 3D anchorage-independent growth,cisplatin resistance,invasion,and bioenergetics of the MPM cell lines tested.We showed that miR-186-5p and TWIST1 levels are anti-correlated in biphasic MPM specimens from TCGA.Conclusion:We unraveled a novel mechanism of action of butein,which attenuated the pro-tumorigenic features of MPM at least through a miR-186-5p-TWIST1 axis.We suggest that those activities converge into the chemo-sensitizing effect of this compound and may be of translational relevance.展开更多
文摘目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄糖调节蛋白78(glucose regulated protein of 78,GRP78)的表达情况;分析临床资料与mH2A和GRP78表达的关系;免疫共沉淀实验检测GRP78和mH2A蛋白的相互作用;Western印迹检测mH2A和GRP78的相关作用及Butein与mH2A之间的作用。Transwell试验检测mH2A的表达对黑色素瘤细胞侵袭能力的影响;划痕试验检测mH2A的表达对黑色素瘤细胞迁移能力的影响;Western印迹检测沉默mH2A后细胞外信号调节激酶1(ERK1)和MAPK/ERK激酶(MEK)蛋白的表达。结果:与癌旁正常组织相比,黑色素瘤组织中mH2A和GRP78表达明显降低(P<0.05);Butein可以增强mH2A的表达水平(P<0.05),mH2A可以调节GRP78蛋白的表达(P<0.05);沉默mH2A促进黑色素瘤A375细胞的侵袭迁移能力,上调MEK和ERK1蛋白的表达。结论:Butein可以增强mH2A靶向GRP78蛋白通过MAPK信号通路抑制黑色素瘤细胞的迁移和侵袭行为。
基金Supported by Grants from the National Research Foundation of Korea,No.R1102452Korea University,No.K1220161by experimental techniques from the Core Laboratory for Convergent Translational Research of College of Medicine
文摘AIM: To evaluate the effects of butein on inflammatory cytokines, matrix metalloproteinase-9(MMP-9), andcolitis in interleukin(IL)-10-/- mice.METHODS: To synchronize colitis, 8- to 10-wk-old IL-10-/- mice were fed pellet-chow containing piroxicam for 2 wk. Subsequently, phosphate-buffered saline or butein(1 mg/kg per day, ip) was injected for 4 wk. Histologic scores, inflammatory cytokines, MMP-9 and phosphorylated signal transducer and activator of transcription 3(p STAT3) expressions were analyzed in IL-10-/- mice and in Colo 205 cells.RESULTS: Butein reduced the colonic inflammatory score by > 50%. Expression levels of IL-6, IL-1β, interferon(IFN)-γ and MMP-9 were decreased in the colons of mice exposed to butein, whereas other inflammatory cytokines(IL-17 A, IL-21 and IL-22) were unchanged. Immunohistochemical staining for p STAT3 and MMP-9 was significantly decreased in the buteintreated groups compared with the controls. Butein inhibited IL-6-induced activation of STAT3 in Colo 205 cells.CONCLUSION: Butein ameliorated colitis in IL-10-/-mice by regulating IL-6/STAT3 and MMP-9 activation.
基金partially supported by an AIRC and Marie Curie Actions-People-COFUND fellowship.
文摘Aim:Malignant pleural mesothelioma is a chemoresistant tumor,and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma(MPM)patients.We obtained the microRNA expression profile of three biphasic-sarcomatoid MPM cell lines to identify commonly expressed microRNAs and evaluate the effect of butein,a chemo-sensitizing compound,on this microRNA subset.Methods:Nanostring-based microRNA profiling and analysis through the ROSALIND platform were employed to identify the commonly modulated microRNAs and their targets.MicroRNA-mimic transfection,Luciferase assay,and Western blotting were employed to show specific perturbation of TWIST1 levels by miR-186-5p.Sphere-forming assays,invasion assay,and metabolic profiling were used to assess the biological consequences of the butein-instigated miR-186-5p-mediated perturbation of TWIST1 levels.TGCA analysis was used to search for the correlation between TWIST1 and miR-186-5p levels in biphasic and epithelioid MPM specimens.Results:We identified a set of perturbed microRNAs,common to three biphasic/sarcomatoid MPM cell lines,after butein treatment.When focusing on miR-186-5p,we unraveled a butein-ignited and miR-186-5p-mediated modulation of TWIST1 levels which affected the 3D anchorage-independent growth,cisplatin resistance,invasion,and bioenergetics of the MPM cell lines tested.We showed that miR-186-5p and TWIST1 levels are anti-correlated in biphasic MPM specimens from TCGA.Conclusion:We unraveled a novel mechanism of action of butein,which attenuated the pro-tumorigenic features of MPM at least through a miR-186-5p-TWIST1 axis.We suggest that those activities converge into the chemo-sensitizing effect of this compound and may be of translational relevance.
