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 ...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 transfection 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±0.009 （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
Background:There is increasing evidence that circular RNAs(circRNAs)play a significant role in pathological processes including tumorigenesis.In contrast to exonic circRNAs,which are the most frequently reported circR...Background:There is increasing evidence that circular RNAs(circRNAs)play a significant role in pathological processes including tumorigenesis.In contrast to exonic circRNAs,which are the most frequently reported circRNAs in cancer so far,the studies of intronic circRNAs have been greatly lagged behind.Here,we aimed to investigate the regulatory role of intronic circRNAs in head and neck squamous cell carcinoma(HNSCC).Methods:We conducted whole-transcriptome sequencing with four pairs of primary tumor tissues and adjacent normal tissues from HNSCC patients.Then,we characterized circGNG7 expression in HNSCC tissues and cell lines and explored its association with the prognosis of HNSCC patients.We also identified interactions between circGNG7 and functional proteins,which alter downstream signaling that regulate HNSCC progression.Results:In this study,we identified a new intronic circRNA,circGNG7,and validated its functional roles in HNSCC progression.CircGNG7 was predominately localized to the cytoplasm,and its expression was downregulated in both HNSCC tissues andCAL27,CAL33,SCC4,SCC9,HN6,and HN30 cells.Low expression of circGNG7 was significantly correlated with poor prognosis in HNSCC patients.Consistent with this finding,overexpression of circGNG7 strongly inhibited tumor cell proliferation,colony formation,in vitro migration,and in vivo tumor growth.Mechanistically,the expression of circGNG7 in HNSCC cells was regulated by the transcription factor SMAD family member 4(SMAD4).Importantly,we discovered that circGNG7 could bind to serine residues 78 and 82 of the functional heat shock protein 27(HSP27),occupying its phosphorylation sites and hindering its phosphorylation,which reduced HSP27-JNK/P38 mitogen-activated protein kinase(MAPK)oncogenic signaling.Downregulation of circGNG7 expression in HNSCC increased HSP27-JNK/P38 MAPK signaling and promoted tumor progression.Conclusions:Our results revealed that a new intronic circRNA,circGNG7,functions as a strong tumor suppressor and that circGNG7/HSP27-JNK/P38 MAPK signaling is a novel mechanism by which HNSCC progression can be controlled.展开更多
The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relationship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was...The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relationship 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 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 hu-man lens epithelial cells (HLB-...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 hu-man lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation in-jury 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 immunohistochem-istry 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 epithe-lial cells.展开更多
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 transfection 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±0.009 （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This study was supported by grants from the National Natural Science Foundation of China(No.81902748 and 81872185)Shanghai Sailing Program(No.19YF1427100)sponsored by Program of Innovative Research Team of High-level Local Universities in Shanghai.
文摘Background:There is increasing evidence that circular RNAs(circRNAs)play a significant role in pathological processes including tumorigenesis.In contrast to exonic circRNAs,which are the most frequently reported circRNAs in cancer so far,the studies of intronic circRNAs have been greatly lagged behind.Here,we aimed to investigate the regulatory role of intronic circRNAs in head and neck squamous cell carcinoma(HNSCC).Methods:We conducted whole-transcriptome sequencing with four pairs of primary tumor tissues and adjacent normal tissues from HNSCC patients.Then,we characterized circGNG7 expression in HNSCC tissues and cell lines and explored its association with the prognosis of HNSCC patients.We also identified interactions between circGNG7 and functional proteins,which alter downstream signaling that regulate HNSCC progression.Results:In this study,we identified a new intronic circRNA,circGNG7,and validated its functional roles in HNSCC progression.CircGNG7 was predominately localized to the cytoplasm,and its expression was downregulated in both HNSCC tissues andCAL27,CAL33,SCC4,SCC9,HN6,and HN30 cells.Low expression of circGNG7 was significantly correlated with poor prognosis in HNSCC patients.Consistent with this finding,overexpression of circGNG7 strongly inhibited tumor cell proliferation,colony formation,in vitro migration,and in vivo tumor growth.Mechanistically,the expression of circGNG7 in HNSCC cells was regulated by the transcription factor SMAD family member 4(SMAD4).Importantly,we discovered that circGNG7 could bind to serine residues 78 and 82 of the functional heat shock protein 27(HSP27),occupying its phosphorylation sites and hindering its phosphorylation,which reduced HSP27-JNK/P38 mitogen-activated protein kinase(MAPK)oncogenic signaling.Downregulation of circGNG7 expression in HNSCC increased HSP27-JNK/P38 MAPK signaling and promoted tumor progression.Conclusions:Our results revealed that a new intronic circRNA,circGNG7,functions as a strong tumor suppressor and that circGNG7/HSP27-JNK/P38 MAPK signaling is a novel mechanism by which HNSCC progression can be controlled.
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 relationship 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.
文摘背景小注意与倒流食道炎( RE )在病人对热吃惊蛋白质 27 的表示( HSP27 )被给予了，并且在食道炎的 HSP27 的重要性的很少研究在学习试图与 RE 在老鼠的食道的织物探索 HSP27 的表示的动物 models.This 被执行了。八十只女 Wistar 老鼠随机被划分成试验性的组 A 和 B 和控制组 C 和 D (在每个组的 n=20 ) 的方法。建立 RE，在二个试验性的组的老鼠收到了幽门和 forestomach 结扎，当在控制的老鼠组织收到的 gastrostomy 和胃的穿孔时，修理。在组 A 和 C 的老鼠被牺牲在外科以后的 7 天，和在组 B 和 D 的老鼠被牺牲在外科以后的 14 天。在组 A 和 B， 10 和 8 只老鼠被病理学的检查与 RE 诊断，分别地(他们在组 A' 和 B' 被包括，分别地) 。在组 C 和 D 的所有更低的食道的纸巾的组织病理学说的诊断是正常的， 20normal 标本随机在每个组与 10 个标本为组 C 和 D' 被选择。宏观、显微镜的食道炎分数在组 A' 和 B' 为标本被估计。更低的食道的纸巾从组 A' ， B' ， C，和 D' 被收集，并且嵌入石蜡的片用纸巾的部分被做。在纸巾的 HSP27 的表示用二拍子的圆舞 streptavidin-peroxidase immunohistochemical 方法被检测。一些收集了纸巾被冻结，并且 HSP27 mRNA 的表情用荧光被检测量的聚合酶链反应(FQ-PCR ) 。中部的宏观、显微镜的食道炎在组 A'(n=10 ) 和 B'(n=8 ) 获得的结果是 1.0 和 1.5，和 2.0 和 2.5 分别地。在在二个组之间的宏观或显微镜的食道炎分数没有重要差别(Z=-0.330， P=0.741；Z=-0.142， P=0.887，分别地) 。染色的 Immunohistochemical 证明 HSP27 在 RE 和控制老鼠在食道的 epithelia 的所有层被表示。FQ-PCR 证明在在 RE 的更低的食道的织物的 HSP27mRNA 层次组织(组 A' 和 B') 比在控制组(组 C' 和 D') 的那些高(Z=-0.249， P=0.001 ) 。在更低的食道的织物的 HSP27 mRNA 表示在组 B' 和 D' 是显著地不同的(Z=-3.027， P=0.002 ) 。并且在严重 RE 的 HSP27 mRNA 表示的层次组织(显微镜的食道炎分数：3 ) 比在温和 RE 组高(显微镜的食道炎分数：1-2 ) 并且控制组(Z=-3.396， P=0.001；Z=-3.855， P < 0.001 ) 。在有 RE 的老鼠的更低的食道的织物的结论 HSP27 mRNA 表示比在正常控制显著地高。尽管倒流是一个坚持的刺激因素，在有 RE 的老鼠的更低的食道的织物的 HSP27 的增加的表示要求加重的食道的损害。
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 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 hu-man lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation in-jury 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 immunohistochem-istry 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 epithe-lial cells.
