The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCL...The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 (78.3 %) and 43 (71.7 %) specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history, whereas HSP 27 expression is not.展开更多
Objective: To clone human cardiac heat shock protein 27 (HSP27) gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods: Full length of HSP27 cDNA which got ...Objective: To clone human cardiac heat shock protein 27 (HSP27) gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods: Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1^+ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable trahsfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H2O2 in H9c2. Results: ①pCDNA3.1^+/HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H2O2 in HSP27 over-expression group and wild type group were 0.396±0.017 vs. 0.390±0.01)9 (p 〉0.05), 0.437±0. 014 vs. 0.416±0.015 (P〈0.05), 0.471±0.018 vs. 0.417±0.009 (P 〈0.001), 0.505±0.030 vs. 0.657± 0.022(P 〈0.001), 0.547 ±0.027 and 0.661 ± 0.011( P 〈 0. 001 ), respectively. ③Apoptosis induced by 150 μmol/L H2O2 in HSP27 over-expression group and wild type group were (10.693± 1.122)% vs. (4.027 ± 1.628)%( P 〈0.01). Conclusion: We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.展开更多
AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glauc...AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glaucoma model and disclose the underlying pathogenesis of glaucoma.METHODS: A total of 115 Wistar rats were randomly divided into 4 groups. Group 1 was the ocular hypertension group by condensing 3 episcleral & limbal veins or episcleral area of right eye(HP group, n=25) and sham operation group with conjunctiva incision without coagulation(n=25). Group 2: HSP27 or dose-matched PBS was injected into the vitreous(V-HSP27 group, n=15;V-PBS group, n=15). Group 3: HSP27 and complete Freund's adjuvant or dosematched PBS was injected subcutaneously into the hind limb accompanied intraperitoneal injection of pertussis toxin [sensitized group(I-HSP27 group), n=15;I-PBS group, n=15)]. Group 4 was normal group without any treatment(n=5). IOPs of the rats were measured before, day 3, weeks 1, 2, 4, 6, and 8 after treatment. Paraffin-embedded sections were prepared for HE staining and RGCs apoptosis were detected by TUNEL. Anti-HSP27 level in serum and CSF were examined by ELISA. RESULTS: IOPs were elevated significantly in HP and V-HSP27, V-PBS groups(P<0.01) and positively related to anti-HSP27 levels in serum and CSFs. Anti-HSP27 levels in serum and CSF were elevated significantly in I-HSP27group compared to other groups(P<0.05). However, the IOPs did not show any relationship with the high-level antiHSP27 in serum and CSFs. RGC apoptosis were all elevated significantly in the HP, V-HSP27, V-PBS and I-HSP27 groups and also positively relative with anti-HSP27 level in serum and CSFs except that high-level of anti-HSP27 in the serum of I-HSP group.CONCLUSION: The increases of anti-HSP27 levels in serum and CSFs both promote IOP escalation and the increase of RGC apoptosis in retina when anti-HSP27 is at low level. The case of high-level anti-HSP27 is opposite and shows protective function in preventing IOP increase and RGC apoptosis.展开更多
Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were f...Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h.Thereafter,the protein expression of HSP-27 and HSP-70 was evaluated by western blotting.Alternatively,HZ-fed cells were cultured up to 72 h and cell viability parameters(survival,apoptosis and necrosis rates) were measured by flow cytometric analysis. Results:HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells,and promoted long-term cell survival without inducing apoptosis.As expected,gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis.Conclusions: Present data show that HZ prevents cell apoptosis and enhances the expression of anli-apoptotic HSP-27 in THP-1 cells,confirming the previous evidences obtained from HZ-fed immunopurified monocytes.Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach,which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.展开更多
The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27) during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HL...The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27) during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation injury group (group B) and sodium salicylate group (group C). Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium (MTT) chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system, Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B (0.2667±0.01414 vs 0.2150±0.01080, P=0.012〈0.05). The average gray value of HSP27 in group B was less than that in group C (P=0.000〈0.05). The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.展开更多
Objective:The aim of the study was to screen differentially expressed proteins between left-and right-sided colon cancers by proteomics techniques and provide molecular genetic basis for oncobiological difference betw...Objective:The aim of the study was to screen differentially expressed proteins between left-and right-sided colon cancers by proteomics techniques and provide molecular genetic basis for oncobiological difference between left-and rightsided colon cancers.Methods:Tissue samples including left-and right-sided colon cancers were collected and preserved in the-80 ℃ refrigeratory.In the first part of our experiment,protein separating was performed by using two-dimensional gel electrophoresis(2-DE) and the images of the gels were acquired by the scanner and then analyzed to find the differentially expression protein-spots in different groups.The peptide mass fingerprintings(PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database.Differentially expression proteins were assayed by RT-PCR,Western blot,and immunohistochemical methods.Results:The 55 differentially expressed protein spots were screened and 23 spots of them were identified.Compared to right-sided colon cancer,15 proteins up-regulated and 8 proteins down-regulated including HSP27 in left-sided colon cancer.HSP27 expressed higher in right-sided than in left-sided colon cancers by RT-PCR,Western blot and immunohistochemical methods.Conclusion:There were differentially expressed proteins between left-and right-sided colon cancers,especially differences in HSP27 expression in mRNA and protein level,which were molecular genetic basis for oncobiological difference between left-and right-sided colon cancers.展开更多
文摘目的分析老年急性脑梗死(acute cerebral infarction,ACI)患者血清氧化低密度脂蛋白(oxidized low density lipoprotein,OX-LDL)、热休克蛋白27(heat shock proteins 27,HSP27)水平与脑水肿严重程度及短期预后的关系。方法回顾性分析2019年1月—2021年9月郑州大学第五附属医院收治的初发老年ACI患者120例(研究组)住院后当天,第3、5、7天的病例资料,按照患者住院当天美国国立卫生院卒中量表(national institute health stroke scale,NIHSS)评分分为轻度组(n=36)、中度组(n=50)、重度组(n=34),于发病后90天,根据改良Rankin量表(modified Rankin Scale,mRS)评分分为预后良好组(n=82)和预后不良组(n=38);另取同期门诊健康体检者120例为对照组。采用酶联免疫吸附试验(ELISA)检测血清OX-LDL、HSP27水平,采用BORN-BE无创脑水肿动态监护仪检测患侧及对照组同侧大脑半球脑电阻抗扰动系数(coefficient of electrical impedance,CEI),采用Pearson法分析血清OX-LDL、HSP27水平与CEI扰动系数的相关性。结果与对照组比较,轻度组、中度组、重度组ACI患者NIHSS评分依次升高,差异有统计学意义(P<0.05)。与对照组比较,轻度组、中度组、重度组ACI患者血清OX-LDL水平、CEI依次增加,差异有统计学意义(P<0.05),均呈先增加后降低趋势,在第5天达到高峰,于第7天开始下降;血清HSP27水平依次降低,差异有统计学意义(P<0.05),随时间变化趋势与OX-LDL相反。Pearson相关性分析结果显示,ACI患者各时间点血清OX-LDL与CEI均呈正相关(P<0.05),血清HSP27水平与CEI均负相关(P<0.05)。与预后良好组比较,预后不良组患者各时间点血清OX-LDL水平升高,差异有统计学意义(P<0.05),随住院时间延长呈先升高后降低趋势,且在第5天达到高峰,第7天开始下降;血清HSP27水平降低,差异有统计学意义(P<0.05),随时间变化趋势与OXLDL相反。结论血清OX-LDL、HSP27水平变化与ACI患者病情、脑水肿严重程度及预后有关,可能对ACI患者脑水肿严重程度及预后评估有一定临床指导意义。
文摘The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relation.ship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 (78.3 %) and 43 (71.7 %) specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history, whereas HSP 27 expression is not.
文摘Objective: To clone human cardiac heat shock protein 27 (HSP27) gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods: Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1^+ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable trahsfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H2O2 in H9c2. Results: ①pCDNA3.1^+/HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H2O2 in HSP27 over-expression group and wild type group were 0.396±0.017 vs. 0.390±0.01)9 (p 〉0.05), 0.437±0. 014 vs. 0.416±0.015 (P〈0.05), 0.471±0.018 vs. 0.417±0.009 (P 〈0.001), 0.505±0.030 vs. 0.657± 0.022(P 〈0.001), 0.547 ±0.027 and 0.661 ± 0.011( P 〈 0. 001 ), respectively. ③Apoptosis induced by 150 μmol/L H2O2 in HSP27 over-expression group and wild type group were (10.693± 1.122)% vs. (4.027 ± 1.628)%( P 〈0.01). Conclusion: We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.
基金Supported by National Natural Science Foundation of China(No.81060077)Key Research and Development Program in Yunnan Province(International Science and Technology Cooperation,No.2017IB001)Science and Technology Planning Project of Guangzhou(No.201604020105)。
文摘AIM: To investigate the relationships between the changes of heat shock protein 27 antibody(anti-HSP27) in serum/cerebrospinal fluid(CSF), intraocular pressure(IOP), retinal ganglion cell(RGC) apoptosis in a rat glaucoma model and disclose the underlying pathogenesis of glaucoma.METHODS: A total of 115 Wistar rats were randomly divided into 4 groups. Group 1 was the ocular hypertension group by condensing 3 episcleral & limbal veins or episcleral area of right eye(HP group, n=25) and sham operation group with conjunctiva incision without coagulation(n=25). Group 2: HSP27 or dose-matched PBS was injected into the vitreous(V-HSP27 group, n=15;V-PBS group, n=15). Group 3: HSP27 and complete Freund's adjuvant or dosematched PBS was injected subcutaneously into the hind limb accompanied intraperitoneal injection of pertussis toxin [sensitized group(I-HSP27 group), n=15;I-PBS group, n=15)]. Group 4 was normal group without any treatment(n=5). IOPs of the rats were measured before, day 3, weeks 1, 2, 4, 6, and 8 after treatment. Paraffin-embedded sections were prepared for HE staining and RGCs apoptosis were detected by TUNEL. Anti-HSP27 level in serum and CSF were examined by ELISA. RESULTS: IOPs were elevated significantly in HP and V-HSP27, V-PBS groups(P<0.01) and positively related to anti-HSP27 levels in serum and CSFs. Anti-HSP27 levels in serum and CSF were elevated significantly in I-HSP27group compared to other groups(P<0.05). However, the IOPs did not show any relationship with the high-level antiHSP27 in serum and CSFs. RGC apoptosis were all elevated significantly in the HP, V-HSP27, V-PBS and I-HSP27 groups and also positively relative with anti-HSP27 level in serum and CSFs except that high-level of anti-HSP27 in the serum of I-HSP group.CONCLUSION: The increases of anti-HSP27 levels in serum and CSFs both promote IOP escalation and the increase of RGC apoptosis in retina when anti-HSP27 is at low level. The case of high-level anti-HSP27 is opposite and shows protective function in preventing IOP increase and RGC apoptosis.
基金supported by University of Torino Intramural Funds to GG and by grants to MP from the Compagnia di San Paolo,Torino,in the context of the Italian Malaria Network
文摘Objective:To investigate the effect of malarial pigment(hemozoin,HZ) on expression of heat shock proteins(HSPs) and cell viability in human monocytes by using a stable cell line(THP-1 cells).Methods:THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h.Thereafter,the protein expression of HSP-27 and HSP-70 was evaluated by western blotting.Alternatively,HZ-fed cells were cultured up to 72 h and cell viability parameters(survival,apoptosis and necrosis rates) were measured by flow cytometric analysis. Results:HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells,and promoted long-term cell survival without inducing apoptosis.As expected,gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis.Conclusions: Present data show that HZ prevents cell apoptosis and enhances the expression of anli-apoptotic HSP-27 in THP-1 cells,confirming the previous evidences obtained from HZ-fed immunopurified monocytes.Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach,which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.
文摘The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27) during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation injury group (group B) and sodium salicylate group (group C). Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium (MTT) chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system, Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B (0.2667±0.01414 vs 0.2150±0.01080, P=0.012〈0.05). The average gray value of HSP27 in group B was less than that in group C (P=0.000〈0.05). The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.
基金Supported by a grant of Foundation Project from Bureau of Health in Hunan Province (No. C2009-010)
文摘Objective:The aim of the study was to screen differentially expressed proteins between left-and right-sided colon cancers by proteomics techniques and provide molecular genetic basis for oncobiological difference between left-and rightsided colon cancers.Methods:Tissue samples including left-and right-sided colon cancers were collected and preserved in the-80 ℃ refrigeratory.In the first part of our experiment,protein separating was performed by using two-dimensional gel electrophoresis(2-DE) and the images of the gels were acquired by the scanner and then analyzed to find the differentially expression protein-spots in different groups.The peptide mass fingerprintings(PMF) was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) and the proteins were identified by data searching in the Mascot-database.Differentially expression proteins were assayed by RT-PCR,Western blot,and immunohistochemical methods.Results:The 55 differentially expressed protein spots were screened and 23 spots of them were identified.Compared to right-sided colon cancer,15 proteins up-regulated and 8 proteins down-regulated including HSP27 in left-sided colon cancer.HSP27 expressed higher in right-sided than in left-sided colon cancers by RT-PCR,Western blot and immunohistochemical methods.Conclusion:There were differentially expressed proteins between left-and right-sided colon cancers,especially differences in HSP27 expression in mRNA and protein level,which were molecular genetic basis for oncobiological difference between left-and right-sided colon cancers.