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Heat shock protein 90 acts as a molecular chaperone in late-phase activation of extracellular signal-regulated kinase 1/2 stimulated by oxidative stress in vascular smooth muscle cells

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Acta Pharmacologica Sinica 2007年第12期28卷 1907-1913页

作者： Dai-hua LIU~2,Hao-yu YUAN~2,Chun-ya CAO,Zhi-ping GAO,Bing-yang ZHU,Hong-lin HUANG~3,Duan-fang LIAO~3division of pharmacoproteomics,Institute of Pharmacy and Pharmacology,School of Life Science and Technology,University of SouthChina,Hengyang 421001,China division of pharmacoproteomics Institute of Pharmacy and Pharmacology School of Life Science and Technology University of South China Hengyang China

Aim:To investigate whether cytosolic heat shock protein 90 (HSP90) acts as amolecular chaperone on the activated extracellular signal-regulated kinase 1/2(ERK1/2) and cell proliferation stimulated by reactive oxygen species (ROS) in ratvascular smooth muscle cells (VSMC).Methods:VSMC were exposed to 1 μmol/LLY83583 (6-anilinoquinoline-5,8-quinolinedione,producer of ROS) for 120 min inthe presence or absence of 5 μmol/L geldanamycin,a specific inhibitor of *** the total,soluble,and insoluble proteins of the cells were ***90,ERK1/2,and phosphor-ERK1/2 in the cell lysate were measured by Western *** interaction of HSP90 and phosphor-ERK1/2 was analyzed by immunoprecipi-tation assay,and the nuclear phosphor-ERK1/2 was measured by Western blot-ting and *** proliferation was tested by cell counting and3-(4,5-dimethylthiazol-2-yl)-3,5-di-phenyltetrazolium bromide (MTT).Results:Thecytosolic HSP90 of VSMC was upregulated by LY83583 in a time-dependent man-ner with the peak at 120 min,which is consistent with the late peak of phosphor-ERK1/*** and Western blotting analyses showed that LY83583increased the interaction of HSP90 with phosphor-ERK 1/2,the phosphor-ERK 1/2level,and the soluble phosphor-ERK1/2 level by 1.8-,2.5-,and 2.9-fold,*** contrast,the insoluble phosphor-ERK1/2 of VSMC was ***,LY83583 treatment promoted the nuclear phosphor-ERK1/2 by 7.6-fold as con-firmed by Western blotting and immunofluorescence ***,cellcounting and the MTT assay showed that LY83583 stimulated VSMC prolifera-tion with the increased expression of HSP90 and levels of soluble and nuclearphosphor-ERK1/*** of geldanamycin antagonized the effect ***:HSP90 could mediate the oxidative stress-stimulated,late-phase activation of ERK1/2 and VSMC proliferation by promoting the ERK1/2phosphorylation,the association of itself with phosphor-ERK1/2,and the solubil-ity

关键词： heat shock protein 90 extracellular signalregulated kinase 1/2 reactive oxygen species vascular smooth muscle cells

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High density lipoprotein 3 inhibits oxidized low density lipoprotein-induced apoptosis via promoting cholesterol efflux in RAW264.7 cells

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Acta Pharmacologica Sinica 2006年第2期27卷 151-157页

作者： Pei JIANG~(2,5) Peng-ke YAN~(2,5) Jian-xiong CHEN~4 Bing-yang ZHU~2 Xiao-yong LEI~2 Wei-dong YIN~(3,6) Duan-fang LIAO~(2,3,6) ~2division of pharmacoproteomics.Institute of Pharmacy and Pharmacology,Nanhua University,Hengyang 421001,China,~3School of Life Science and Technology,Nanhua University,Hengyang 421001,China,~4Department of Pathology,Vanderbilt University Medical Center Nashville.TN 37232.USA division of pharmacoproteomics Institute of Pharmacy and Pharmacology Nanhua University Hengyang China Department of Pathology Vanderbilt University Medical Center Nashville USA School of Life Science and Technology Nanhua University Hengyang China

Aim:To investigate the protective effect of high density lipoprotein 3（HDL3）onoxidized low density lipoprotein（ox-LDL）-induced apoptosis in RAW264.7 cells,Methods:RAW264.7 cells were exposed to 50 mg/L ox-LDL for various durationsup to 48h,and apoptosis was detected using Hoechst 33258 staining and flowcytometric *** cholesterol levels were detected by high performanceliquid chromatography,cholesterol efflux was determined by Tritium labeling,andthe cellular lipid droplets were assayed by oil red O ***:Treatmentwith 50mg/L ox-LDL for 12,24,and 48h increased the apoptotic rate of RAW264,7 cells in a time-dependent *** peak apoptotic rate（47.7%）was observedafter 48h ***3at various concentrations（50mg/L,100mg/L,and 200mg/L）inhibited the ox-LDL（50mg/L for 48h）-mediated apoptosis that was accom-panied by an increased rate of intracellular cholesterol efflux,and decreased totalcholesterol levels in cells in a concentration-dependent *** ofcholesterol efflux by brefeldin decreased the protective effect of HDL3on ox-LDL-induced *** of the cholesterol efflux effected by another choles-terol acceptor,β-cyclodextrin,led to a dramatic decrease in the apoptotic rate ***:HDL3antagonizes ox-LDL-induced apoptosis in RAW264.7cells,through reducing the accumulation of toxic cholesterol.

关键词： cholesterol efflux high density lipoprotein-3 apoptosis oxidized low density lipoprotein

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Effect of Daxx on cholesterol accumulation in hepatic cells

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World Journal of Gastroenterology 2008年第3期14卷 435-440页

作者： Qin-Hui Tuo Lei Liang Bing-Yang Zhu Xuan Cao Duan-Fang Liao Department of Pharmacology School of Pharmaceutical Science Central South UniversityChangsha 410078 Hunan Province China division of pharmacoproteomics School of Life Science and Technology University of South China Western Changsheng Road#28 Hengyang 421001 Hunan Province China

AIM： To study the effect of Daxx on cholesterol accumulation in hepatic cells. METHODS： Sprague Dawley （SD） rats were fed a normal or high fat diet for 6 wk, and serum lipids and Daxx expression of hepatic tissues were measured by immunoblot assays. HepG2 cells were transfected with the pEGFP-C1/Daxx or pEGFP-C1 plasmid. Cells stably transfected with Daxx were identified by RTPCR analysis. Total cholesterol levels were determined by high performance liquid chromatography. Activated- SREBP and caveolin-1 were assayed by western blotting. RESULTS： Hepatic Daxx protein was higher in normal rats than in high fat diet-fed rats. Noticeable negative correlations were seen between Daxx and LDL-C （γ=-7.56, ρ=0.018）, and between Daxx and TC （γ=-9.07, ρ= 0.01）, respectively. The total cholesterol of HepG2/GFP-Daxx cells was lower than that of control cells or HepG2/GFP cells （9.28±0.19 vs 14.36± 4.45 or 13.94±2.62, both P 〈 0.05）. Furthermore, in HepG2/GFP cells, the expression of activated SREBP was lower than that of control cells, whereas caveolin-1 expression was higher. CONCLUSION： Overexpression of Daxx in HepG2 cells decreased intracellular cholesterol accumulation, which might be associated with inhibition of SREBP activity and an increase in caveolin-1 expression.

关键词： Daxx Cholesterol Hepatic cells Sterol regulatory element-binding protein Caveolin-1

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Current status of predictive biomarkers for neoadjuvant therapy in esophageal cancer

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World Journal of Gastrointestinal Pathophysiology 2014年第3期5卷 322-334页

作者： Norihisa Uemura Tadashi Kondo Department of Gastroenterological Surgery Aichi Cancer Center Hospital division of pharmacoproteomics National Cancer Center Research Institute

Neoadjuvant therapy has been proven to be extremely valuable and is widely used for advanced esophageal cancer. However, a significant proportion of treated patients(60%-70%) does not respond well to neoadjuvant treatments and develop severe adverse effects. Therefore, predictive markers for individualization of multimodality treatments are urgently needed in esophageal cancer. Recently, molecular biomarkers that predict the response to neoadjuvant therapy have been explored in multimodal approaches in esophageal cancer and successful examples of biomarker identification have been reported. In this review, promising candidates for predictive molecular biomarkers developed by using multiple molecular approaches are reviewed. Moreover, treatment strategies based on the status of predicted biomarkers are discussed, while considering the international differences in the clinical background. However, in the absence of adequate treatment options related to the results of the biomarker test, the usefulness of these diagnostic tools is limited and new effective therapies for biomarker-identified nonresponders to cancer treatment should be concurrent with the progress of predictive technologies. Further improvement in the prognosis of esophageal cancer patients can be achieved through the introduction of novel therapeutic approaches in clinical practice.

关键词： Esophageal cancer Neoadjuvant therapy Response prediction Molecular biomarker Chemoradiation

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