Objective:Hepatocellular carcinoma(HCC),the main type of liver cancer,has a high morbidity and mortality,and a poor prognosis.RNA helicase DDX5,which acts as a transcriptional co-regulator,is overexpressed in most mal...Objective:Hepatocellular carcinoma(HCC),the main type of liver cancer,has a high morbidity and mortality,and a poor prognosis.RNA helicase DDX5,which acts as a transcriptional co-regulator,is overexpressed in most malignant tumors and promotes cancer cell growth.Heat shock protein 90(HSP90)is an important molecular chaperone in the conformational maturation and stabilization of numerous proteins involved in cell growth or survival.Methods:DDX5 m RNA and protein expression in surgically resected HCC tissues from 24 Asian patients were detected by quantitative real-time PCR and Western blot,respectively.The interaction of DDX5-HSP90 was determined by molecular docking,immunoprecipitation,and laser scanning confocal microscopy.The autophagy signal was detected by Western blot.The cell functions and signaling pathways of DDX5 were determined in 2 HCC cell lines.Two different murine HCC xenograft models were used to determine the function of DDX5 and the therapeutic effect of an HSP90 inhibitor.Results:HSP90 interacted directly with DDX5 and inhibited DDX5 protein degradation in the AMPK/ULK1-regulated autophagy pathway.The subsequent accumulation of DDX5 protein induced the malignant phenotype of HCC by activating theβ-catenin signaling pathway.The silencing of DDX5 or treatment with HSP90 inhibitor both blocked in vivo tumor growth in a murine HCC xenograft model.High levels of HSP90 and DDX5 protein were associated with poor prognoses.Conclusions:HSP90 interacted with DDX5 protein and subsequently protected DDX5 protein from AMPK/ULK1-regulated autophagic degradation.DDX5 and HSP90 are therefore potential therapeutic targets for HCC.展开更多
Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from ...Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from cerebral ischemia/reperfusion and to determine whether these effects are associated with the expression level of HSPA5 in the ischemic penumbra.Methods A total of 72 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group[middle cerebral artery occlusion(MCAO)-Ex,n=18],ischemia and sedentary group(MCAO-St,n=18),sham-surgery and exercise group(Sham-Ex,n=18),or sham-surgery and sedentary group(Sham-St,n=18).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 min,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 min once a day for 5 consecutive days.After reperfusion,the motor function of the rats was scored by the Bederson neurological function test,balance beam test,and screen test.Nissl staining was conducted to assess morphological and structural change of nerve cells in the ischemic penumbra.The reverse transcription-quantitative polymerase chain reaction was applied to detect the mRNA expression of HSPA5.Western blot analysis was conducted to determine the protein expression of HSPA5.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was carried out in the ischemic penumbra after MCAO.Results Rats receiving early treadmill exercise had lower Bederson neurological function,balance beam,and screen test scores on the 3rd,7th,and 14th days after MCAO;in addition,more neurons survived in the ischemic penumbra after MCAO,and higher mRNA and protein expression of HSPA5 and fewer TUNEL-positive stained cells were observed.Conclusion Our study demonstrated that early aerobic exercise can improve neurological function recovery after ischemia/reperfusion.Furthermore,the increased level of HSPA5 in the ischemic penumbra might be one of the mechanisms of enhanced neurological function recovery.展开更多
AIM: To investigate putative biological damage caused by GSM mobile phone frequencies by assessing electromagnetic fields during mobile phone working. METHODS: Neuron-like cells, obtained by retinoicacid-induced diffe...AIM: To investigate putative biological damage caused by GSM mobile phone frequencies by assessing electromagnetic fields during mobile phone working. METHODS: Neuron-like cells, obtained by retinoicacid-induced differentiation of human neuroblastoma SH-SY5Y cells, were exposed for 2 h and 4 h to microwaves at 1800 MHz frequency bands. RESULTS: Cell stress response was evaluated by MTT assay as well as changes in the heat shock protein expression (Hsp20, Hsp27 and Hsp70) and caspase-3 activity levels, as biomarkers of apoptotic pathway. Under our experimental conditions, neither cell viability nor Hsp27 expression nor caspase-3 activity was significantly changed. Interestingly, a significant decrease in Hsp20 expression was observed at both times of exposure, whereas Hsp70 levels were significantly increased only after 4 h exposure. CONCLUSION: The modulation of the expression of Hsps in neuronal cells can be an early response to radiofrequency microwaves.展开更多
OBJECTIVE： TO summarize the relationship between heat shock protein 70 （HSP70） and cerebra ischemia. DATA SOURCES： An online search of Medline database was undertaken to identify relevant articles published in Eng...OBJECTIVE： TO summarize the relationship between heat shock protein 70 （HSP70） and cerebra ischemia. DATA SOURCES： An online search of Medline database was undertaken to identify relevant articles published in English from January 1980 to December 2005 by using the keywords of ＂heat shock protein 70, ischemia＂. Meanwhile, Chinese relevant articles published from January 2000 to December 2005 were searched in China National Knowledge Infrastructure （CNKI） database and Chinese Journal of Clinical Rehabilitation with the keywords of ＂heat shock protein 70, cerebral ischemia＂ in Chinese. STUDY SELECTION ： More than 100 related articles were screened, and 29 references mainly about HSP70 and cerebral ischemia were selected, including basic and clinical researches. As to the articles with similar content, those published in the authoritative journals in recent 3 years were preferential. DATA EXTRACTION： A total of 29 articles were collected and classified according to the structure, function and clinical application of HSP70. Among them, 1 article is about the structure of HSP70, 27 about the relationship between HSP70 and cerebral ischemia, and 2 about the clinical application of HSP70. DATA SYNTHESIS： HSP70 is one of the most conservative proteins during biological evolution. Experiments in cerebral ischemia revealed that HSP70 expression was time-dependent, also correlated with the injured site and severity. The cerebral ischemia induced HSP70 gene expression in hippocampus of gerbil had protection to tolerance of fatal ischemic injury for neurons. The increase of HSP70 expression may be one of the endogenous protective mechanisms during cerebral ischemia, and can effectively alleviate cerebral ischemia. Thus HSP70 protein and HSP70 mRNA have been taken as important indexes extensively applied in the basic study of cerebral ischemia by some scholars abroad. CONCLUSION： HSP70 plays a protective role in cerebral ischemia, and a deeper research into the biological function of HSP70 will provide a new way for the therapy of cerebral ischemia.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
基金funding support from the National Natural Science Foundation of China(Grant Nos.81672467,81702773,81702389,and 81672368)the Major National R&D Project(Grant Nos.2018ZX10723204,2018ZX10302205,and 2018ZX09J18107)the Natural Science Foundation of Beijing(Grant No.7172207)。
文摘Objective:Hepatocellular carcinoma(HCC),the main type of liver cancer,has a high morbidity and mortality,and a poor prognosis.RNA helicase DDX5,which acts as a transcriptional co-regulator,is overexpressed in most malignant tumors and promotes cancer cell growth.Heat shock protein 90(HSP90)is an important molecular chaperone in the conformational maturation and stabilization of numerous proteins involved in cell growth or survival.Methods:DDX5 m RNA and protein expression in surgically resected HCC tissues from 24 Asian patients were detected by quantitative real-time PCR and Western blot,respectively.The interaction of DDX5-HSP90 was determined by molecular docking,immunoprecipitation,and laser scanning confocal microscopy.The autophagy signal was detected by Western blot.The cell functions and signaling pathways of DDX5 were determined in 2 HCC cell lines.Two different murine HCC xenograft models were used to determine the function of DDX5 and the therapeutic effect of an HSP90 inhibitor.Results:HSP90 interacted directly with DDX5 and inhibited DDX5 protein degradation in the AMPK/ULK1-regulated autophagy pathway.The subsequent accumulation of DDX5 protein induced the malignant phenotype of HCC by activating theβ-catenin signaling pathway.The silencing of DDX5 or treatment with HSP90 inhibitor both blocked in vivo tumor growth in a murine HCC xenograft model.High levels of HSP90 and DDX5 protein were associated with poor prognoses.Conclusions:HSP90 interacted with DDX5 protein and subsequently protected DDX5 protein from AMPK/ULK1-regulated autophagic degradation.DDX5 and HSP90 are therefore potential therapeutic targets for HCC.
基金This study was funded by Basic Research Projects in Shanxi Province(No.2015021178)Start-up Funds for Doctoral Research at Shanxi Datong University(No.2014-B-01).
文摘Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from cerebral ischemia/reperfusion and to determine whether these effects are associated with the expression level of HSPA5 in the ischemic penumbra.Methods A total of 72 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group[middle cerebral artery occlusion(MCAO)-Ex,n=18],ischemia and sedentary group(MCAO-St,n=18),sham-surgery and exercise group(Sham-Ex,n=18),or sham-surgery and sedentary group(Sham-St,n=18).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 min,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 min once a day for 5 consecutive days.After reperfusion,the motor function of the rats was scored by the Bederson neurological function test,balance beam test,and screen test.Nissl staining was conducted to assess morphological and structural change of nerve cells in the ischemic penumbra.The reverse transcription-quantitative polymerase chain reaction was applied to detect the mRNA expression of HSPA5.Western blot analysis was conducted to determine the protein expression of HSPA5.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was carried out in the ischemic penumbra after MCAO.Results Rats receiving early treadmill exercise had lower Bederson neurological function,balance beam,and screen test scores on the 3rd,7th,and 14th days after MCAO;in addition,more neurons survived in the ischemic penumbra after MCAO,and higher mRNA and protein expression of HSPA5 and fewer TUNEL-positive stained cells were observed.Conclusion Our study demonstrated that early aerobic exercise can improve neurological function recovery after ischemia/reperfusion.Furthermore,the increased level of HSPA5 in the ischemic penumbra might be one of the mechanisms of enhanced neurological function recovery.
文摘AIM: To investigate putative biological damage caused by GSM mobile phone frequencies by assessing electromagnetic fields during mobile phone working. METHODS: Neuron-like cells, obtained by retinoicacid-induced differentiation of human neuroblastoma SH-SY5Y cells, were exposed for 2 h and 4 h to microwaves at 1800 MHz frequency bands. RESULTS: Cell stress response was evaluated by MTT assay as well as changes in the heat shock protein expression (Hsp20, Hsp27 and Hsp70) and caspase-3 activity levels, as biomarkers of apoptotic pathway. Under our experimental conditions, neither cell viability nor Hsp27 expression nor caspase-3 activity was significantly changed. Interestingly, a significant decrease in Hsp20 expression was observed at both times of exposure, whereas Hsp70 levels were significantly increased only after 4 h exposure. CONCLUSION: The modulation of the expression of Hsps in neuronal cells can be an early response to radiofrequency microwaves.
基金a grant from Sci-entific and Technological Devel-opmental Program of Beijing E-ducation Committee, No.KM200510025004
文摘OBJECTIVE： TO summarize the relationship between heat shock protein 70 （HSP70） and cerebra ischemia. DATA SOURCES： An online search of Medline database was undertaken to identify relevant articles published in English from January 1980 to December 2005 by using the keywords of ＂heat shock protein 70, ischemia＂. Meanwhile, Chinese relevant articles published from January 2000 to December 2005 were searched in China National Knowledge Infrastructure （CNKI） database and Chinese Journal of Clinical Rehabilitation with the keywords of ＂heat shock protein 70, cerebral ischemia＂ in Chinese. STUDY SELECTION ： More than 100 related articles were screened, and 29 references mainly about HSP70 and cerebral ischemia were selected, including basic and clinical researches. As to the articles with similar content, those published in the authoritative journals in recent 3 years were preferential. DATA EXTRACTION： A total of 29 articles were collected and classified according to the structure, function and clinical application of HSP70. Among them, 1 article is about the structure of HSP70, 27 about the relationship between HSP70 and cerebral ischemia, and 2 about the clinical application of HSP70. DATA SYNTHESIS： HSP70 is one of the most conservative proteins during biological evolution. Experiments in cerebral ischemia revealed that HSP70 expression was time-dependent, also correlated with the injured site and severity. The cerebral ischemia induced HSP70 gene expression in hippocampus of gerbil had protection to tolerance of fatal ischemic injury for neurons. The increase of HSP70 expression may be one of the endogenous protective mechanisms during cerebral ischemia, and can effectively alleviate cerebral ischemia. Thus HSP70 protein and HSP70 mRNA have been taken as important indexes extensively applied in the basic study of cerebral ischemia by some scholars abroad. CONCLUSION： HSP70 plays a protective role in cerebral ischemia, and a deeper research into the biological function of HSP70 will provide a new way for the therapy of cerebral ischemia.
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。
