Prostate cancer preferentially metastasizes to the bone. However, the underlying molecular mechanisms are still unclear. To explore the effects of a bone-mimicking microenvironment on PC3 prostate cancer cell growth a...Prostate cancer preferentially metastasizes to the bone. However, the underlying molecular mechanisms are still unclear. To explore the effects of a bone-mimicking microenvironment on PC3 prostate cancer cell growth and metastasis, we used osteoblast differentiation medium（ODM; minimal essential medium alpha supplemented with L-ascorbic acid） to mimic the bone microenvironment. PC3 cells grown in ODM underwent epithelial-mesenchymal transition and showed enhanced colony formation, migration, and invasion abilities compared to the cells grown in normal medium. PC3 cells grown in ODM showed enhanced metastasis when injected in mice. A screening of signaling pathways related to invasion and metastasis revealed that the NF-κB pathway was activated, which could be reversed by Bay 11-7082, a NF-κB pathway inhibitor. These results indicate that the cells in different culture conditions manifested significantly different biological behaviors and the NF-κB pathway is a potential therapeutic target for prostate cancer bone metastasis.展开更多
动脉性肺动脉高压(pulmonary arterial hypertension,PAH)的发生、发展与骨形态发生蛋白受体II型(bone morphogeneticprotein receptor type II,BMPRII)编码基因的遗传突变和核因子κB(nuclear factorκB,NF-κB)通路介导的炎症反应密...动脉性肺动脉高压(pulmonary arterial hypertension,PAH)的发生、发展与骨形态发生蛋白受体II型(bone morphogeneticprotein receptor type II,BMPRII)编码基因的遗传突变和核因子κB(nuclear factorκB,NF-κB)通路介导的炎症反应密切相关。本文旨在研究NF-κB通路抑制剂对脂多糖(lipopolysaccharide,LPS)诱导的肺动脉内皮细胞损伤的作用。用1μg/mL的LPS处理人肺动脉内皮细胞,用免疫印迹和qPCR检测BMPRII和白介素8(interleukin-8,IL-8)的表达水平。腹腔注射野百合碱(monocrotaline,MCT)建立大鼠PAH模型,用免疫荧光染色法检测肺动脉内皮细胞BMPRII和IL-8的表达情况,检测模型大鼠心脏血流动力学变化和肺血管重构情况。结果显示,LPS可引起人肺动脉内皮细胞BMPRII的表达下调和IL-8的表达上调,NF-κB抑制剂BAY11-7082(10μmol/L)可逆转LPS的作用。在MCT-PAH大鼠模型中,肺动脉内皮细胞BMPRII表达下调,IL-8表达上调,右心室/(左心室+室间隔)重量比值[weight ratio of right ventricle to left ventricle plus septum,RV/(LV+S)]和右心室收缩压(rightventricular systolic pressure,RVSP)显著升高,心输出量(cardiac output,CO)和三尖瓣环收缩期位移(tricuspid annular plane systolicexcursion,TAPSE)明显降低,肺血管壁明显增厚,连续21天腹腔注射BAY11-7082(5 mg/kg)可逆转上述变化。以上结果提示,LPS通过NF-κB信号通路下调BMPRII的表达水平,促进PAH的发生、发展,因此NF-κB信号通路可作为PAH潜在治疗靶点。展开更多
This study explored the effects of nuclear factor-kappa B(NF-κB)inhibitor Bay 11-7082 on Fas/FasL system and Fas-mediated apoptosis in cell line HL-60 cells.The mRNA and protein levels of Fas,FasL,and X-linked inhibi...This study explored the effects of nuclear factor-kappa B(NF-κB)inhibitor Bay 11-7082 on Fas/FasL system and Fas-mediated apoptosis in cell line HL-60 cells.The mRNA and protein levels of Fas,FasL,and X-linked inhibitor of apoptosis protein(XIAP)were detected by reverse transcription-polymerase chain reaction(RT-PCR)and flow cytometry(FCM);the level of sFasL was evaluated by enzyme-linked immunosorbent assay(ELISA);and apoptosis was determined by FCM.After treatment with Bay 11-7082,the mRNA and protein levels of FasL and XIAP in HL-60 cells were significantly lower than in the controls(P<0.05),but the mRNA and protein levels of Fas and sFasL did not change significantly(P>0.05).Apoptotic rate of HL-60 cells treated with Bay 11-7082 was significantly higher than in the controls(P<0.05).Therefore,we conclude that Bay 11-7082 can enhance Fas-mediated apoptosis in HL-60 cells by downregulating FasL and XIAP levels.展开更多
基金supported by National Natural Science Foundation of China (NSFC) (81272415 and 81171993)NSFC Key Project (81130046)+1 种基金Guangxi Key Projects (2013GXNSFEA053004)Guangxi Projects (2014GXNSFDA118030)
文摘Prostate cancer preferentially metastasizes to the bone. However, the underlying molecular mechanisms are still unclear. To explore the effects of a bone-mimicking microenvironment on PC3 prostate cancer cell growth and metastasis, we used osteoblast differentiation medium（ODM; minimal essential medium alpha supplemented with L-ascorbic acid） to mimic the bone microenvironment. PC3 cells grown in ODM underwent epithelial-mesenchymal transition and showed enhanced colony formation, migration, and invasion abilities compared to the cells grown in normal medium. PC3 cells grown in ODM showed enhanced metastasis when injected in mice. A screening of signaling pathways related to invasion and metastasis revealed that the NF-κB pathway was activated, which could be reversed by Bay 11-7082, a NF-κB pathway inhibitor. These results indicate that the cells in different culture conditions manifested significantly different biological behaviors and the NF-κB pathway is a potential therapeutic target for prostate cancer bone metastasis.
基金This work was supported by the National Natural Science Foundation of China(No.81670054)the Innovation Project of Chinese Academy of Medical Sciences(No.CIFMS 2016-I2M-4-003)。
文摘动脉性肺动脉高压(pulmonary arterial hypertension,PAH)的发生、发展与骨形态发生蛋白受体II型(bone morphogeneticprotein receptor type II,BMPRII)编码基因的遗传突变和核因子κB(nuclear factorκB,NF-κB)通路介导的炎症反应密切相关。本文旨在研究NF-κB通路抑制剂对脂多糖(lipopolysaccharide,LPS)诱导的肺动脉内皮细胞损伤的作用。用1μg/mL的LPS处理人肺动脉内皮细胞,用免疫印迹和qPCR检测BMPRII和白介素8(interleukin-8,IL-8)的表达水平。腹腔注射野百合碱(monocrotaline,MCT)建立大鼠PAH模型,用免疫荧光染色法检测肺动脉内皮细胞BMPRII和IL-8的表达情况,检测模型大鼠心脏血流动力学变化和肺血管重构情况。结果显示,LPS可引起人肺动脉内皮细胞BMPRII的表达下调和IL-8的表达上调,NF-κB抑制剂BAY11-7082(10μmol/L)可逆转LPS的作用。在MCT-PAH大鼠模型中,肺动脉内皮细胞BMPRII表达下调,IL-8表达上调,右心室/(左心室+室间隔)重量比值[weight ratio of right ventricle to left ventricle plus septum,RV/(LV+S)]和右心室收缩压(rightventricular systolic pressure,RVSP)显著升高,心输出量(cardiac output,CO)和三尖瓣环收缩期位移(tricuspid annular plane systolicexcursion,TAPSE)明显降低,肺血管壁明显增厚,连续21天腹腔注射BAY11-7082(5 mg/kg)可逆转上述变化。以上结果提示,LPS通过NF-κB信号通路下调BMPRII的表达水平,促进PAH的发生、发展,因此NF-κB信号通路可作为PAH潜在治疗靶点。
文摘This study explored the effects of nuclear factor-kappa B(NF-κB)inhibitor Bay 11-7082 on Fas/FasL system and Fas-mediated apoptosis in cell line HL-60 cells.The mRNA and protein levels of Fas,FasL,and X-linked inhibitor of apoptosis protein(XIAP)were detected by reverse transcription-polymerase chain reaction(RT-PCR)and flow cytometry(FCM);the level of sFasL was evaluated by enzyme-linked immunosorbent assay(ELISA);and apoptosis was determined by FCM.After treatment with Bay 11-7082,the mRNA and protein levels of FasL and XIAP in HL-60 cells were significantly lower than in the controls(P<0.05),but the mRNA and protein levels of Fas and sFasL did not change significantly(P>0.05).Apoptotic rate of HL-60 cells treated with Bay 11-7082 was significantly higher than in the controls(P<0.05).Therefore,we conclude that Bay 11-7082 can enhance Fas-mediated apoptosis in HL-60 cells by downregulating FasL and XIAP levels.
