热休克蛋白70(heat shock protein 70,Hsp70)是一种在应激条件下诱导表达的伴侣蛋白,能帮助细胞内新生蛋白折叠,蛋白细胞内运输、蛋白装配和降解,并能在应激状态下维持蛋白质构象。研究表明,Hsp70与许多疾病有关,如癌症、神经退行性疾...热休克蛋白70(heat shock protein 70,Hsp70)是一种在应激条件下诱导表达的伴侣蛋白,能帮助细胞内新生蛋白折叠,蛋白细胞内运输、蛋白装配和降解,并能在应激状态下维持蛋白质构象。研究表明,Hsp70与许多疾病有关,如癌症、神经退行性疾病、异源移植的排斥、感染等,有望成为新的药物作用靶点。本文综述了Hsp70的结构、家族成员、协同伴侣(co-chaperones)成员及其功能研究进展,并介绍Hsp70抑制剂的研究现况。展开更多
目的:探讨DNAJ热激蛋白家族B8基因(DNAJ heat shock protein family member B8,DNAJB8)在肺腺癌组织中的表达及其对肺腺癌细胞侵袭转移的作用和可能机制。方法:收集四川省人民医院2006年至2008年的肺腺癌手术切除标本102例,通过免疫组...目的:探讨DNAJ热激蛋白家族B8基因(DNAJ heat shock protein family member B8,DNAJB8)在肺腺癌组织中的表达及其对肺腺癌细胞侵袭转移的作用和可能机制。方法:收集四川省人民医院2006年至2008年的肺腺癌手术切除标本102例,通过免疫组织化学(IHC)实验检测DNAJB8在肺腺癌组织中的表达情况,并分析其与临床病理参数及预后的关系。构建DNAJB8稳定敲低的肺腺癌A549细胞和DNAJB8稳定过表达的肺腺癌H1299细胞,CCK-8实验检测DNAJB8敲降或过表达对肺腺癌细胞增殖的影响,Transwell法检测其对肺腺癌细胞侵袭能力的影响,Western blotting检测其对肺腺癌细胞内侵袭相关蛋白MMP-2、MMP-9和ERK表达及活性的影响。裸鼠尾静脉注射DNAJB8稳定敲低的A549细胞构建肺腺癌转移模型,观察DNAJB8对A549细胞体内转移能力的影响。结果:DNAJB8在肺腺癌组织中的表达明显高于正常肺组织,其高表达与患者的淋巴结转移及TNM分期呈正相关,并且预示着患者有较差的预后(均P<0.01)。DNAJB8在A549细胞中的表达量高于H1299细胞,下调DNAJB8表达能够抑制A549细胞的侵袭[(41±3)vs(192±11)个,P<0.01],而上调DNAJB8的表达能够促进H1299细胞的侵袭[(235±14)vs(25±4)个,P<0.01]。实验组肺腺癌转移模型裸鼠肺脏肿瘤结节数显著低于对照组[(5±1)vs(17±3)个,P<0.01]。A549细胞中DNAJB8敲降后,MMP-2、MMP-9表达量和p-ERK水平均显著下降(均P<0.01);而H1299细胞高表达DNAJB8后,MMP-2、MMP-9表达量和p-ERK水平均显著升高(均P<0.01)。结论:DNAJB8能够促进肺腺癌的侵袭和转移,其机制可能与MEK/Erk信号通路有关。展开更多
目的:探讨热休克蛋白5(heat shock protein family A member 5,HSPA5)对青蒿琥酯诱导人肝癌SMMC-7721细胞株化疗敏感性的影响。方法:用不同浓度青蒿琥酯处理SMMC-7721细胞,CCK-8法检测细胞活性,筛选最佳实验浓度。将SMMC-7721细胞按以...目的:探讨热休克蛋白5(heat shock protein family A member 5,HSPA5)对青蒿琥酯诱导人肝癌SMMC-7721细胞株化疗敏感性的影响。方法:用不同浓度青蒿琥酯处理SMMC-7721细胞,CCK-8法检测细胞活性,筛选最佳实验浓度。将SMMC-7721细胞按以下分组处理:对照组、青蒿琥酯组、青蒿琥酯+去铁胺组,用流式细胞术检测细胞内脂质来源活性氧水平;试剂盒检测细胞内丙二醛水平。用包装HSPA5干扰或过表达质粒的慢病毒感染SMMC-7721细胞,qRT-PCR和蛋白质印迹法分别测定转染后HSPA5 mRNA和蛋白表达;CCK-8法检测细胞活性,试剂盒检测细胞内丙二醛水平。结果:青蒿琥酯浓度为20μmol/L时,SMMC-7721细胞达到半数致死量,为最佳实验浓度;流式细胞术结果显示,青蒿琥酯处理的细胞内脂质来源活性氧水平、丙二醛水平明显升高,去铁胺可抑制青蒿琥酯导致的细胞内脂质来源活性氧和丙二醛升高(均P <0. 05);经过青蒿琥酯处理的HSPA5干扰组细胞活性水平明显低于未干扰组,丙二醛水平明显高于未干扰组(均P <0. 05)。结论:干扰HSPA5可能增强人肝癌SMMC-7721细胞株对青蒿琥酯的化疗敏感性。展开更多
Wilson's disease(WD) is an autosomal recessive disorder of copper metabolism, caused by mutations in the ATP7 B gene. A clear demand for novel WD treatment strategies has emerged. Although therapies using zinc sal...Wilson's disease(WD) is an autosomal recessive disorder of copper metabolism, caused by mutations in the ATP7 B gene. A clear demand for novel WD treatment strategies has emerged. Although therapies using zinc salts and copper chelators can effectively cure WD, these drugs exhibit limitations in a substantial pool of WD patients who develop intolerance and/or severe side effects. Several lines of research have indicated intriguing potential for novel strategies and targets for development of new therapies. Here, we review these new approaches, which comprise correction of ATP7 B mutants and discovery of new compounds that circumvent ATP7B-deficiency, as well as cell and gene therapies. We also discuss whether and when these new therapeutic strategies will be translated into clinical use, according to the key requirements for clinical trials that remain to be met. Finally, we discuss the hope for the current rapidly developing research on molecular mechanisms underlying WD pathogenesis and for the related potential therapeutic targets to provide a solid foundation for the next generation of WD therapies that may lead to an effective, tolerable and safe cure.展开更多
目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细...目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。展开更多
Fibrolamellar hepatocellular carcinoma(FLC)is a rare but aggressive liver cancer of children that occurs predominantly in teenagers without a history of liver disease.Surgical resection remains the only therapeutic op...Fibrolamellar hepatocellular carcinoma(FLC)is a rare but aggressive liver cancer of children that occurs predominantly in teenagers without a history of liver disease.Surgical resection remains the only therapeutic option,and the recurrence rate is extremely high(>50%within 3 years).A newly discovered chromosomal deletion that occurs in the majority of FLCs generates a novel kinase fusion between DnaJ heat shock protein family member B1(DNAJB1)and protein kinase cAMP-activated catalytic subunit alpha(PRKACA)(DNAJB1-PRKACA).Despite its high penetrance and apparent specificity for FLC,the oncogenic role of this fusion event remains unclear.In this review article,we discuss the histology,presentation and diagnosis,current treatment,and roles of the DNAJB1-PRKACA as well as research models contributing to our understanding of this disease.展开更多
文摘热休克蛋白70(heat shock protein 70,Hsp70)是一种在应激条件下诱导表达的伴侣蛋白,能帮助细胞内新生蛋白折叠,蛋白细胞内运输、蛋白装配和降解,并能在应激状态下维持蛋白质构象。研究表明,Hsp70与许多疾病有关,如癌症、神经退行性疾病、异源移植的排斥、感染等,有望成为新的药物作用靶点。本文综述了Hsp70的结构、家族成员、协同伴侣(co-chaperones)成员及其功能研究进展,并介绍Hsp70抑制剂的研究现况。
文摘目的:探讨DNAJ热激蛋白家族B8基因(DNAJ heat shock protein family member B8,DNAJB8)在肺腺癌组织中的表达及其对肺腺癌细胞侵袭转移的作用和可能机制。方法:收集四川省人民医院2006年至2008年的肺腺癌手术切除标本102例,通过免疫组织化学(IHC)实验检测DNAJB8在肺腺癌组织中的表达情况,并分析其与临床病理参数及预后的关系。构建DNAJB8稳定敲低的肺腺癌A549细胞和DNAJB8稳定过表达的肺腺癌H1299细胞,CCK-8实验检测DNAJB8敲降或过表达对肺腺癌细胞增殖的影响,Transwell法检测其对肺腺癌细胞侵袭能力的影响,Western blotting检测其对肺腺癌细胞内侵袭相关蛋白MMP-2、MMP-9和ERK表达及活性的影响。裸鼠尾静脉注射DNAJB8稳定敲低的A549细胞构建肺腺癌转移模型,观察DNAJB8对A549细胞体内转移能力的影响。结果:DNAJB8在肺腺癌组织中的表达明显高于正常肺组织,其高表达与患者的淋巴结转移及TNM分期呈正相关,并且预示着患者有较差的预后(均P<0.01)。DNAJB8在A549细胞中的表达量高于H1299细胞,下调DNAJB8表达能够抑制A549细胞的侵袭[(41±3)vs(192±11)个,P<0.01],而上调DNAJB8的表达能够促进H1299细胞的侵袭[(235±14)vs(25±4)个,P<0.01]。实验组肺腺癌转移模型裸鼠肺脏肿瘤结节数显著低于对照组[(5±1)vs(17±3)个,P<0.01]。A549细胞中DNAJB8敲降后,MMP-2、MMP-9表达量和p-ERK水平均显著下降(均P<0.01);而H1299细胞高表达DNAJB8后,MMP-2、MMP-9表达量和p-ERK水平均显著升高(均P<0.01)。结论:DNAJB8能够促进肺腺癌的侵袭和转移,其机制可能与MEK/Erk信号通路有关。
文摘目的:探讨热休克蛋白5(heat shock protein family A member 5,HSPA5)对青蒿琥酯诱导人肝癌SMMC-7721细胞株化疗敏感性的影响。方法:用不同浓度青蒿琥酯处理SMMC-7721细胞,CCK-8法检测细胞活性,筛选最佳实验浓度。将SMMC-7721细胞按以下分组处理:对照组、青蒿琥酯组、青蒿琥酯+去铁胺组,用流式细胞术检测细胞内脂质来源活性氧水平;试剂盒检测细胞内丙二醛水平。用包装HSPA5干扰或过表达质粒的慢病毒感染SMMC-7721细胞,qRT-PCR和蛋白质印迹法分别测定转染后HSPA5 mRNA和蛋白表达;CCK-8法检测细胞活性,试剂盒检测细胞内丙二醛水平。结果:青蒿琥酯浓度为20μmol/L时,SMMC-7721细胞达到半数致死量,为最佳实验浓度;流式细胞术结果显示,青蒿琥酯处理的细胞内脂质来源活性氧水平、丙二醛水平明显升高,去铁胺可抑制青蒿琥酯导致的细胞内脂质来源活性氧和丙二醛升高(均P <0. 05);经过青蒿琥酯处理的HSPA5干扰组细胞活性水平明显低于未干扰组,丙二醛水平明显高于未干扰组(均P <0. 05)。结论:干扰HSPA5可能增强人肝癌SMMC-7721细胞株对青蒿琥酯的化疗敏感性。
文摘Wilson's disease(WD) is an autosomal recessive disorder of copper metabolism, caused by mutations in the ATP7 B gene. A clear demand for novel WD treatment strategies has emerged. Although therapies using zinc salts and copper chelators can effectively cure WD, these drugs exhibit limitations in a substantial pool of WD patients who develop intolerance and/or severe side effects. Several lines of research have indicated intriguing potential for novel strategies and targets for development of new therapies. Here, we review these new approaches, which comprise correction of ATP7 B mutants and discovery of new compounds that circumvent ATP7B-deficiency, as well as cell and gene therapies. We also discuss whether and when these new therapeutic strategies will be translated into clinical use, according to the key requirements for clinical trials that remain to be met. Finally, we discuss the hope for the current rapidly developing research on molecular mechanisms underlying WD pathogenesis and for the related potential therapeutic targets to provide a solid foundation for the next generation of WD therapies that may lead to an effective, tolerable and safe cure.
文摘目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。
文摘Fibrolamellar hepatocellular carcinoma(FLC)is a rare but aggressive liver cancer of children that occurs predominantly in teenagers without a history of liver disease.Surgical resection remains the only therapeutic option,and the recurrence rate is extremely high(>50%within 3 years).A newly discovered chromosomal deletion that occurs in the majority of FLCs generates a novel kinase fusion between DnaJ heat shock protein family member B1(DNAJB1)and protein kinase cAMP-activated catalytic subunit alpha(PRKACA)(DNAJB1-PRKACA).Despite its high penetrance and apparent specificity for FLC,the oncogenic role of this fusion event remains unclear.In this review article,we discuss the histology,presentation and diagnosis,current treatment,and roles of the DNAJB1-PRKACA as well as research models contributing to our understanding of this disease.
