Survivin和热休克蛋白27在胰腺癌组织中的表达及关系

目的:探讨Survivin和热休克蛋白27(HSP27)在胰腺癌组织中的表达及二者的相关性。方法:用免疫组织化学法检测59例胰腺癌及癌旁组织中Survivin与HSP27的表达,比较其阳性表达率及Survivin表达与HSP27表达的关系。结果:Survivin在胰腺癌中阳性表达率为74.6%,而在癌旁组织中未见表达(P<0.01);Survivin表达与胰腺癌的分化程度、临床分期及淋巴结转移无相关性;HSP27在胰腺癌中的表达率为66.1%,明显高于癌旁组织22.8%(P<0.01),并且与胰腺癌的分化程度相关,但与临床分期及淋巴结转移无关;胰腺癌组织中Survivin与HSP27的表达有相关性。结论:Survivin和HSP27在胰腺癌中过度表达,提示二者在胰腺癌的发生发展中起重要作用;Survivin和HSP27通过抗凋亡机制协同参与胰腺癌的发生。     

谷氨酰胺抑制人外周血单个核细胞细胞因子表达的机制

目的 探讨谷氨酰胺抑制人外周血单个核细胞(PBMCs)细胞因子表达的分子机制.方法 Ficoll密度梯度离心法提取健康志愿者新鲜PBMCs,将分离得到的细胞分为两部分.第一部分细胞分为六部分,分别用不同浓度的Gln(0、8、15 mmol/L)预处理0.5 h及2.0 h后,再用10 g/L内毒素刺激4.0 h,留取细胞及上清.第二部分细胞分为A、B、C三组,将热休克蛋白(HSP)阻断剂Quercetin(200 μmol/L)加入A组及B组中,作用0.5 h;再将谷氨酰胺8 mmol/L加入A组和C组中;1.0 h后在三组中均加入10 g/L内毒素刺激4.0 h,留取细胞及上清.酶联免疫吸附(ELISA)测定上清中PBMCs肿瘤坏死因子-α(TNF-α)和白介素-10(IL-10)的表达,免疫印迹(Western Blot)技术检测细胞内HSP70的表达情况.对指标检测结果进行统计学分析.结果 8 mmol/L及15 mmol/L谷氨酰胺预处理0.5 h后能够明显增加HSP70的表达(P<0.05),8 mmol/L谷氨酰胺预处理2.0 h后能明显增加HSP70的表达(P<0.05).同时TNF-α和IL-10表达受抑(P<0.05);使用HSP70阻断剂Quercetin后,HSP70表达量明显下降,同时TNF-α和IL-10表达增加(P<0.05).结论 谷氨酰胺抑制内毒素刺激下PBMCs细胞因子的表达与HSP70介导有关.     

热休克蛋白在肝细胞癌形成、增殖、复发、转移及抗肿瘤免疫中的作用研究

【摘要】 热休克蛋白(heat shock proteins, HSPs)是一类结构高度保守的蛋白质,主要参与蛋白的转运、折叠、维持蛋白质自身稳定及免疫应答。大量研究表明,HSPs中多个成员如HSP27、HSP70及HSP90,在肝细胞癌(hepatocellular carcinoma, HCC)中呈过表达。本文就HSPs在肝细胞癌的形成、增殖、复发、转移及抗肿瘤免疫的作用进行综述。     

HSP27 和claudin-10在结直肠癌中的表达及其临床意义

目的：探讨HSP27和claudin-10在结直肠癌中的表达及其临床意义。 方法：应用免疫组化法检测50例结直肠癌组织、25例结直肠腺瘤组织、50例正常结直肠黏膜组织中HSP27和claudin-10的表达,并分析结直肠癌中两者表达的关系以及与结直肠癌患者临床病理因素及预后的关系。 结果：在结直肠癌组织、结直肠腺瘤组织、正常结直肠黏膜中HSP27阳性表达率分别为54.0%、20.0%、16.0%,差异有统计学意义（P<0.001）;claudin-10阳性表达率72.0%、56.0%、54.0%,差异无统计学意义（P>0.05）;结直肠癌中HSP27与claudin-10的表达呈正相关（r=0.318,P=0.024）。单因素分析显示,HSP27的阳性表达率与与淋巴结转移有关（P<0.05）;claudin-10的阳性表达率与肿瘤直径、浸润深度及淋巴结转移有关（均P<0.05）。生存分析显示,HSP27和claudin-10阳性表达患者平均生存期明显低于各自的阴性表达患者（均P<0.05）。多因素分析显示,HSP27、claudin-10表达及淋巴结转移是影响结直肠癌患者预后的独立风险因素（均P<0.05）。 结论：HSP27在结直肠癌中组织中表达上调,并可能与claudin-10协同在结直肠癌转移中起重要作用。     

热休克蛋白70疫苗在消化道肿瘤研究中的进展

目的综述热休克蛋白70(heat shock protein70,HSP70)疫苗在消化道肿瘤中的研究进展。方法对近年来的相关文献进行分析。结果HSP70能与肿瘤特异性抗原结合,形成HSP70-多肽复合物;它作为"分子伴侣"参与肿瘤免疫作用,激活机体特异和非特异免疫反应。结论HSP70的应用在消化道肿瘤的预防和治疗中显示出诱人的前景。     

shRNA介导的热休克蛋白70敲低对人结肠癌细胞在裸鼠体内生长的影响

目的:探讨热休克蛋白70(HSP70)敲低对结肠癌细胞在裸鼠体内生长的影响。方法:构建两种干扰HSP70的shRNA(HSP70 shRNA-1和HSP70 shRNA-2)质粒表达载体,分别转染到结肠癌HT29细胞,并以空质粒转染(阴性对照)和未转染的HT29细胞(空白对照)为对照,用RT-PCR和Western blot方法检测各组HT29细胞HSP70基因和蛋白的表达;将HSP70shRNA-2转染、空质粒转染及未转染的HT29细胞分别接种至裸鼠皮下建立移3组植瘤模型,观察肿瘤生长情况,3周后剥离瘤块,用HE染色、免疫组化和TUNEL法分别检测移植瘤组织形态学、增殖细胞核抗原(PCNA)的表达及细胞凋亡情况。结果:RT-PCR和Western blot结果显示HSP70 shRNA-1和HSP70 shRNA-2均能明显抑制HT29细胞HSP70基因和蛋白的表达,且HSP70 shRNA-2的抑制作用更为明显,空质粒转染对HSP70的表达无明显影响;与空白对照组比较,HSP70 shRNA-2转染组裸鼠移植瘤生长明显较明显减慢(P<0.01),瘤组织中心出现明显坏死,PCNA表达明显降低,细胞凋亡率明显增加(P<0.01),而空质粒转染组无明显上述改变(P>0.05)。结论:HSP70沉默能抑制结肠癌HT29细胞裸鼠移植瘤的生长,促进细胞凋亡,HSP70可能是治疗结肠癌有效靶点。     

Ezrin shRNA联合HSP70对骨肉瘤细胞凋亡和增殖的影响

 目的 探讨Ezrin基因沉默联合热休克蛋白（heat shock protein, HSP）70诱导的免疫杀伤对骨肉瘤细胞增殖和凋亡的影响。方法 采用人成骨肉瘤MG63细胞系作为研究对象,将人HSP70和Ezrin-shRNA的DNA片段分别克隆至含CMV和pU6双启动子的表达载体pGFP-V-RS中构建含HSP70和Ezrin-shRNA的真核表达载体pGFP-V-RS-shRNA和pGFP-V-RS-shRNA-HSP70。重组质粒分别转染入细胞,筛选出稳定表达的细胞克隆。荧光显微镜观察细胞形态及转染效果;荧光定量RT-PCR及蛋白印迹western blot检测稳定转染细胞株Ezrin和HSP70基因及蛋白水平的变化;采用MTT、流式细胞术检测细胞增殖、凋亡能力变化,Western blot检测凋亡及周期相关蛋白表达量变化;MTT法检测HSP70刺激产生特异性细胞毒性T淋巴细胞（CTL）对靶肿瘤细胞的杀伤作用。结果 荧光图像及基因和蛋白表达分析证实,首次成功构建同时沉默Ezrin和过表达HSP70的特异性载体。较单纯沉默Ezrin,同时过表达HSP70会在一定程度上减弱沉默Ezrin蛋白促进细胞凋亡和抑制增殖的效果,但与对照细胞相比,M63细胞凋亡率仍明显上升,自11.01%±0.22%上升至24.28%±0.50%,增殖速度则自395.14%±2.24%减少至310.00%±2.83%。另外,Western blot检测发现Ezrin-shRNA可促进凋亡基因Bax的表达,相反可降低抗凋亡基因Bcl-2和细胞周期蛋白Cyclin D1的表达。而Ezrin-shRNA/HSP70组较Ezrin-shRNA组促Bax表达和降低Bcl-2、细胞周期蛋白Cyclin D1表达有所减弱,但较阴性对照组仍有明显效果。MG63细胞的CTL细胞毒杀伤效应显示各效靶浓度下CT+IL-2+HSP70组的杀伤活性高于CT+IL-2组,最高达56.33%±1.95%。结论 同时沉默Ezrin、过表达HSP70既可以促进骨肉瘤细胞凋亡抑制其增殖,并利用HSP70诱导的CTL,增强对靶肿瘤细胞的杀伤作用。     

热休克蛋白47与肾脏纤维化

热休克蛋白47(HSP47)是胶原特异性“分子伴侣”，可能在不同器官的纤维化进程中起重要作用。本文综述了HSP47的基因结构、分子特点、功能及在肾脏纤维化中的作用。     

热休克蛋白70在大鼠急性打击损伤脊髓中的表达

目的观察急性打击损伤大鼠脊髓组织中热休克蛋白70（heat shock protein70，HSP 70）的表达。方法65只大鼠随机分为3组：正常对照组5只、手术对照组和脊髓损伤组各30只，采用改良Allen法建立脊髓损伤动物模型。手术对照组和脊髓损伤组分别于处置后的2h、6h、12h、24h、48h、72h这6个时间点各采集5只大鼠的脊髓。应用免疫组织化学染色方法观察不同时点各组脊髓中热休克蛋白的表达变化。结果在大鼠正常胸段脊髓中没有基础性HSP70的表达。打击造成急性脊髓损伤后2h，脊髓组织内出现HSP70的表达，损伤后24～48h脊髓组织中HSP70染色达到高峰，并维持至损伤后72h。结论在遭遇损伤性刺激后，脊髓组织在随后的2～72h内HSP70的表达明显增加；HSP70可能在阻止脊髓的继发性损伤方面发挥一定的作用。     

热休克蛋白70 mRNA在膀胱癌中的表达及与细胞凋亡的关系

目的　探讨热休克蛋白 70 (HSP 70 )mRNA在膀胱移行细胞癌中的表达及与细胞凋亡的关系。　方法　应用原位杂交法对 6 0例膀胱癌组织中HSP 70mRNA进行检测 ,原位DNA末端标记法 (TUNEL)检测细胞凋亡情况。　结果　 6 0例标本中 ,HSP 70mRNA阳性表达率为 5 6 .7% (34/ 6 0 )。HSP 70mRNA表达随肿瘤分级增高而增加 (P <0 .0 5 ) ;G1,G2 和G3 各级间两两比较 ,HSP 70mRNA表达差异也有显著性意义 (P <0 .0 5 )。随着肿瘤分期增高 ,HSP 70mRNA表达随之增加 (P <0 .0 5 ) ;Ⅰ ,Ⅱ和Ⅲ各期间两两比较 ,差异有显著性意义 (P <0 .0 5 )。肿瘤细胞凋亡指数 (AIs)随恶性程度的增高显著降低 (P <0 .0 5 ) ;HSP 70mRNA表达率和AIs呈负相关 (rs=- 0 .6 85 ,P <0 .0 5 )。　结论　HSP 70mRNA表达随膀胱癌恶性程度增加显著增高 ,细胞凋亡则显著降低 ,提示HSP 70表达能够抑制膀胱癌细胞的凋亡。     

热休克蛋白家族与肿瘤的相关性及其临床应用的研究进展

热休克蛋白是一组应激蛋白,在多数肿瘤细胞中表达增高,并参与肿瘤的生长、侵袭、转移等,与患者预后密切相关。本文就HSPs的特点、功能、与肿瘤发生发展的关系及临床应用分别进行介绍,深入了解和认识热休克蛋白与肿瘤之间发生作用的机制和规律,对研究肿瘤的防治策略具有重要意义。     

胃癌组织中HSP60和HSP70的表达及其意义

目的研究胃癌组织中HSP60和HSP70的表达及其意义.方法将胃癌组织石蜡包埋切片,以ABC免疫组织化学法染色进行观察.结果50例胃癌HSP60和HSP70阳性率分别为56%和64%,高分化和未发生淋巴结转移的胃癌HSP60和HSP70阳性率均明显地低于未分化和有淋巴结转移的胃癌.结论HSP60和HSP70的表达可能与胃癌临床病理特征、生物学行为和预后有较密切的关系,两者可能是反映胃癌生物学行为和预后的重要标记物.     

Increased level of HSP27 but not of HSP72 in human heart allografts in relation to acute rejection

BACKGROUND: Increased expression of heat shock proteins (HSPs) was assumed during cardiac allograft rejection. To find evidence for this in man, we quantified HSP27 and HSP72 in cardiac allograft biopsies. METHODS: In parallel to histological assessment of rejection, HSP27 was quantified by Western blotting in a total of 43 biopsies sampled from 3 patients. HSP72 was analyzed in parallel in 30 of the 43 cases. For comparison, HSPs were analyzed in myocardium. RESULTS: HSP27 was significantly higher in rejecting cardiac allografts than in non-rejecting allografts and non-failing myocardium (1.52 +/- 0.25 vs. 0.83 +/- 0.11 vs. 0.50 +/- 0.05 microg/mg protein). Similarity for HSP72 (6.27 +/- 1.54 vs. 4.06 +/- 1.03 vs. 6.27 +/- 0.76 microg/mg protein) was not found. CONCLUSION: For the first time in humans with cardiac allograft rejection, increased expression of HSP27, which could be important for cardiac self-protection, was demonstrated. For the lack of increased HSP72 expression, the influence of the cyclosporine A treatment was discussed.     

热休克蛋白47重组质粒对病理性瘢痕的体内干预研究

目的 利用RNAi技术通过热休克蛋白47重组质粒(HSP47siRNA)和脂质体的混合液对裸鼠病理性瘢痕动物模型的体内干预,分析HSP47基因在病理性瘢痕生成中的意义.方法 构建裸鼠病理性瘢痕动物模型,第16天腹腔麻醉后实验组裸鼠在病理性瘢痕内注射质粒、脂质体混和液0.25 ml,对照组裸鼠腹腔注射PBS液0.25 ml,原笼饲养,7 d回收标本,分别作mRNA水平、胶原蛋白水平以及免疫组化检测.结果 对照组与实验组总胶原含量分别为(91.71±1.24)%和(82.12±4.79)%;实时荧光PCR检测HSP47mRNA表达对照组和实验组分别为1 042 862.01±604 194.36和306 123.68±105 857.08;Ⅰ型胶原蛋白mRNA表达对照组和实验组分别为10 228 614.70±2 532 879.04和6 011 841.97±2 886 897.17;对照组和实验组体积分别为(255.60±21.34)mm~3和(132.99±24.06)mm~3,上述指标检测结果组间比较差异均有统计学意义(P<0.05);而这些变化在增生性瘢痕组并没有出现,增生性瘢痕组胶原的变化、体积的变化均无统计学意义.结论 采用RNAi技术,经过HSP47siRNA表达载体特异性沉默病理性瘢痕中HSP47基因表达后,瘢痕疙瘩中胶原蛋白的合成和分泌均能得到明显抑制,提示HSP47基因促进瘢痕疙瘩生成,并为抑制瘢痕疙瘩提供了新的靶点.     

热休克蛋白90α在人胃癌组织中的表达及临床病理分析

目的研究热休克蛋白90α在人胃癌组织和正常胃黏膜组织中的表达,并探讨其表达与胃癌的发生、发展及生物学行为的关系。方法收集临床46例胃癌组织和相应的正常胃黏膜组织标本,应用SP免疫组化法检测HSP90α的表达情况,结合患者的临床病理资料分析检测结果。结果HSP90α在胃癌组织中的阳性表达率为82．6％,明显高于正常胃黏膜组织中的表达（21．7％）;胃癌组织中HSP90α阳性表达率与患者年龄、性别、肿瘤大小及分化程度等无明显相关性。HSP90α在肿瘤浸润至肌层或越过肌层者的阳性表达率（89．5％）明显高于肿瘤局限于黏膜或黏膜下层者（50．0％）。淋巴结转移7个以上（即N2、N3期）的病例中HSP90α阳性表达率明显高于淋巴结转移为N0和N1的病例（96．2％vs65．0％,P〈0．05）;TNM分期Ⅲ、Ⅳ期阳性表达率也要高于Ⅰ、Ⅱ期（92．9％VS66．7％,P〈0．05）。结论HSP90α在胃癌组织中呈现高表达,并且在浸润深的肿瘤或淋巴结转移多和越晚期的胃癌组织中表达越高。提示HSP90α可作为胃癌预后判定监测指标。     

ATPase inhibition by omeprazole reveals role of heat shock proteins on testicular torsion

Testicular torsion leads ischaemic injury and generates reactive oxygen species. Reactive oxygen species triggers lipid peroxidation, protein degradation and DNA damage. These biochemical processes trigger tissue damage. Heat shock proteins (HSPs) are important in spermatogenesis, and this work elucidates role of HSPs at the testicular torsion–detorsion process. A proton-pump inhibitor, omeprazole, tested to reveal the drug's curative effect since HSP functions through ATP hydrolysis. Thirty-two male Wistar Albino rats were divided into four groups: sham, control, omeprazole and serum physiologic groups. Right testis was torsed, while left ones remained untorsed. Protein peroxidation, DNA damage and lipid hydroperoxide levels as well as HSP expression were measured. Further, the effects were visualised with histopathologic imaging. HSP expression increases at the torsed right testis compared to the contralateral testis. Although HSP70 and HSP90 help antioxidant enzymes to keep their native structure, their anti-apoptotic properties accelerate the tissue damage. Omeprazole a proton-pump inhibitor employed to impair electron transfer chain and to inhibit HSP ATPase function. Omeprazole effectively inhibits HSPs and alleviates lipid peroxidation and DNA damage levels both at molecular and at tissue level, and the drug has profound curative effect on testicular torsion recovery.     

热休克蛋白在子宫颈癌中的表达及其意义

目的探讨热休克蛋白(heat shock proteins,HSPs)在子宫颈癌组织中的表达情况及意义。方法采用PT—PCRH和免疫印迹法检测HSPs在子宫颈癌组织及正常子宫组织中的蛋白和mRNA的表达水平。结果癌组织中HSP70、HSP86及αB晶状体蛋白表达较正常组织明显增多(P<0.05),且HSP70表达最高(P<0.01)。结论 HSP70在子宫颈癌组织中表达显著增高。     

Expression of HSP70 in healing wounds of diabetic and nondiabetic mice.

BACKGROUND: Heat shock proteins (HSPs) stabilize intracellular processes of cells under stress. Little is known about the role of HSPs in wound healing, or whether their expression is altered by systemic disease. The focus of this study was to examine the local heat shock response to wounding in diabetic mice. METHODS: Congenitally diabetic and phenotypically normal mice underwent standardized full-thickness cutaneous wounding. Mice were sacrificed at sequential time points and the wound beds excised. Tissues underwent immunohistochemical (IHC) and RT-PCR analyses for inducible HSP70. RESULTS: HSP70 protein expression in the wound bed by IHC peaked at 24 h in the nondiabetic mice. Expression of HSP70 was delayed in the diabetic mice until Day 3, which correlates with the clinical delay in healing seen in this model. The protein was especially prominent in the epithelium and in inflammatory cells migrating into the granulation tissue matrix. RT-PCR demonstrated upregulation of HSP70 mRNA within 12 h after wounding, lasting until Day 3, and decreasing thereafter in both the nondiabetic and the diabetic animals. CONCLUSION: Cutaneous wounding produces a HSP response in inflammatory cells, and expression of inducible HSP70 is delayed in diabetic mice. This delay may be related to the impaired inflammatory response of diabetics, and may contribute to impaired wound healing. The wound may be a continuing source of the heat shock response in inflammatory cells after injury.     

Pharmacological induction of HSP27 attenuates intimal hyperplasia in vivo.

OBJECTIVES: intimal hyperplasia (IH) is a major cause of re-stenosis post-vascular intervention. Induction of heat shock proteins (HSPs), by thermal pre-conditioning, reduces IH. Our aim was to investigate the effect of the pharmacological HSP inducer herbimycin A on IH in the rat carotid balloon injury model. Materials and Methods: thirty male Sprague-Dawley rats were randomized into three groups. All groups underwent balloon injury to the left carotid artery. Stress proteins were induced 18 h pre-operatively by heat shock or herbimycin A. Two weeks post-operatively, animals were sacrificed and carotid intima/media area ratio (I/M ratio) calculated using computerized planimetry. Neo-intimal proliferation was assessed immunohistochemically with PCNA (proliferating cell nuclear antigen). Western blot and immunohistochemistry for arterial HSP70 and HSP27 were performed. RESULTS: heat stress and herbimycin significantly reduced the I/M ratio (p < 0.05 vs balloon injury alone). Neo-intimal proliferation was significantly reduced in the heat stress and herbimycin groups (p < 0.05 vs balloon injury alone). Heat stress induced arterial HSP70 and HSP27. Herbimycin A increased arterial HSP27. CONCLUSION: herbimycin A significantly attenuates IH after balloon injury. HSP27 may be the HSP involved in mediating this response. Pharmacological inducers of HSPs may have a therapeutic role to play in preventing re-stenosis post-vascular intervention.