Heat shock protein 90 promotes RNA helicase DDX5 accumulation and exacerbates hepatocellular carcinoma by inhibiting autophagy

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.

Heat shock protein 90 is a potential therapeutic target for ameliorating skeletal muscle abnormalities in Parkinson's disease

Previous studies have confirmed that heat shock protein 90 overexpression can lead to dopami- nergic neuronal death. This study was designed to further investigate what effects are produced by heat shock protein 90 after endurance exercise training. Immunohistochemistry results showed that exercise training significantly inhibited heat shock protein 90 overexpression in the soleus and gastrocnemius in Parkinson＇s disease rats, which is a potential therapeutic target for ameliorating skeletal muscle abnormalities in Parkinso^s disease.

Research Progress of Heat Shock Protein 90 and Hepatocellular Carcinoma

Heat shock protein (HSP) is a kind of protein that mainly acts as a molecular chaperone to participate in the synthesis and folding of proteins, maintain the spatial conformation of proteins and protect cells from damage and other important biological functions. HSP90 plays an important role in maintaining molecular chaperone structure, regulating cell cycle and apoptosis, coordinating hormone signal transduction and promoting wound healing. And HSP90 also plays an important role in the occurrence and progression of tumors. In recent years, HSP90 inhibitors have made some achievements in molecular targeted therapy for malignant tumors, but further research is needed in clinical application. In this paper, the research status of the relationship between hepatocellular carcinoma targeted by heat shock protein 90 was reviewed.

Glucocorticoid receptor and heat shock protein 90 in peripheral blood mononuclear cells from asthmatics

Objective To investigate the expression of glucocerticoid receptor (GR) and heat shock protein 90(HSP90) mRNA in peripheral blood mononuclear cells (PBMCs) from steroid-sensitive (SS), steroiddependent (SD) and steroid-resistant (SR) asthmatics patients, and to evaluate the role of GR and HSP90in the pathogenesis of SR.Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the expressions of GR and HSP90 mRNA in PBMC stimulated with IL-2 and/or IL-4 from 10 normal volunteers,10 SS, 5 SD and 6 SR patients.Results The expressions of GR and HSP90 mRNA were the highest in PBMC from SR patients (0.730±0.171 and 1.122±0.165, respectively) compared with the normals (P＜0.05). The second was from SS patients (0.359±0.350 and 0.885±0.250, respectively). The lowest was from the SD patients (0.017±0.008 and 0.078 ± 0.039, respectively). The ratio of HSP90/GR in SR was significantly lower than that in the others, and it suggested that the expression of HSP90 mRNA was not sufficient in this group of patients. When PBMC from SS, SD and SR was incubated with IL-2 or IL-4 alone, no changes in GR and HSP90 mRNA expression were observed. While incubated with combination of IL-2 and IL-4, a significantly higher expression of GR mRNA was observed in all asthmatics, and a significantly higher expression of HSP90 mRNA was observed only in SS and SD patients.Conclusion The lowering of HSP90/GR ratio may be one of the causes of SR. IL-2 and IL-4 may play roles in the imbalance of HSP90/GR.

Discovery and development of natural heat shock protein 90 inhibitors in cancer treatment

Heat shock protein 90(Hsp90)is a highly conserved molecular chaperone that plays a vital role in the signal transduction of cancers.Hsp90 inhibitors are able to inhibit Hsp90 or the complex of Hsp90 and co-chaperones resulting in the degradation of Hsp90-dependent client proteins through the ubiqui tina tion-proteasome pathway,thereby leading to the growth inhibition of tumor cells.This review will briefy discuss the molecular structure and biological function of Hsp90,and focus on a summary of recent progress in the development and testing of natural Hsp90 inhibitors and their different means by which they interact with Hsp90.

Heat shock protein 90 inhibition by 17-DMAG lessens disease in the MRL/Ipr mouse model of systemic lupus erythematosus

Elevated expression of heat shock protein 90 （HSP90） has been found in kidneys and serum of systemic lupus erythematosus （SLE） patients and MRLIMp-FasIprIFasJpr（MRLIIpr） autoimmune mice. We investigated if inhibition of HSP90 would reduce disease in MRL/ Ipr mice. In vitro, pretreatment of mesangial cells with HSP90 inhibitor Geldanamycin prior to immune-stimulation showed reduced expression of IL-6, IL-12 and NO. In vivo, we found HSP90 expression was elevated in MRL/Ipr kidneys when compared to C57BL/6 mice and MRIJIpr mice treated with HSP90 inhibitor 17-DMAG. MRIJIpr mice treated with 17-DMAG showed decreased proteinuria and reduced serum anti-dsDNA antibody production. Glomerulonephritis and glomerular IgG and C3 were not significantly affected by administration of 17-DMAG in MRIJIpr. 17-DMAG increased CD8＋ T cells, reduced double-negative T cells, decreased the CD4/CD8 ratio and reduced follicular B cells. These studies suggest that HSP90 may play a role in regulating T-cell differentiation and activation and that HSP90 inhibition may reduce inflammation in lupus.

Amino acids essential for the interaction between cellular heat shock protein 90 and a Kaposi's sarcoma-associated herpesvirus-encoded protein kinase ORF36

Dear Editor,Protein-protein interactions（PPIs）often play important roles in biological processes（Zhang et al.,2016）.The split Renilla luciferase complementation assay（SRLCA）is one of the methods in studying PPIs.SRLCA is based on the complementation of the N-terminal domains of Renilla luciferase（LN）and C-terminal domains of Renilla luciferase （LC） non-functional halves of Renilla luciferase fused to possibly interacting proteins and emit luminescence （Deng et al., 2011; Jiang et al., 2010） （Supplementary Figure S1A）.

Temperature stress response of heat shock protein 90(Hsp90)in the clam Paphia undulata

The Heat shock proteins(HSPs)are a group of molecular chaperones that play a crucial role in cell response to various stresses.A full-length cDNA of the heat shock protein 90(PuHsp90)was cloned and sequenced from the clam Paphia undulata.Phylogenetic analysis revealed that PuHsp90 grouped with the Hsp90 from other metazoan species.Expression of PuHsp90 was highly detected in the gonad,followed by digest gland,gills and heart but was found poorly expressed in mantle,adductor muscle and hemocytes.After a heat shock stress at 32℃up-regulation of PuHsp90 was detected in the mantle,adductor muscle,gills and hemocytes.Maximal expression levels occurred at 4 h after the heat shock and up-regulation is indicative of protein denaturation and of an increase in energy consumption.In contrast after the heat shock,PuHsp90 was continuously down-regulated in the digestive gland and in the gonad suggesting modifications of the biochemical pathways and energy budgets involved in the synthesis of other protein,such as catalase and of other Hsp proteins.These results reveal that PuHsp90 may play an important role in the clam response to a temperature stress.

Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism

<abstract>Aim: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism. Methods: In 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis. Results: The sperm motility declined significantly after cryopreservation (P<0.01). The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1±3.2 and 243.0±21.6, respectively, while those after thawing were 23.2±2.5 and 105.7±28.5, respectively. Both parameters were decreased significantly (P<0.01). No HSP90 was found in the seminal plasma before and after cryopreservation. Conclusion: HSP90 in human spermatozoa was decreased substantially after cryopreservation. This may result from protein degradation, rather than leakage into the seminal plasma.

Hepatitis C virus inhibitor synergism suggests multistepinteractions between heat-shock protein 90 and hepatitis Cvirus replication

AIM: To address the effect of heat-shock protein 90(HSP90) inhibitors on the release of the hepatitis C virus(HCV), a cell culture-derived HCV(JFH1/HCVcc) from Huh-7 cells was examined.METHODS: We quantified both the intracellular and extracellular(culture medium) levels of the components(RNA and core) of JFH-1/HCVcc. The intracellular HCV RNA and core levels were determined after the JFH1/HCVcc-infected Huh-7 cells were treated with radicicol for 36 h. The extracellular HCV RNA and core protein levels were determined from the medium of the last 24 h of radicicol treatment. To determine the possible role of the HSP90 inhibitor in HCV release, we examined the effect of a combined application of low doses of the HSP90 inhibitor radicicol and the RNA replication inhibitors cyclosporin A(Cs A) or interferon. Finally, we statistically examined the combined effect of radicicoland Cs A using the combination index(CI) and graphical representation proposed by Chou and Talalay.RESULTS: We found that the HSP90 inhibitors had greater inhibitory effects on the HCV RNA and core protein levels measured in the medium than inside the cells. This inhibitory effect was observed in the presence of a low level of a known RNA replication inhibitor(Cs A or interferon-α). Treating the cells with a combination of radicicol and cyclosporin A for 24 h resulted in significant synergy(CI < 1) that affected the release of both the viral RNA and the core protein. CONCLUSION: In addition to having an inhibitory effect on RNA replication, HSP90 inhibitors may interfere with an HCV replication step that occurs after the synthesis of viral RNA, such as assembly and release.

Combined Detection of Serum Heat Shock Protein-90<i>α</i>and Prostate Specific Antigen for Prostate Cancer Diagnosis

Objective: To explore the relationship between heat shock protein-90α (HSP-90α) and occurrence of prostate cancer, and clinical value of combined detection of serum HSP-90α and prostate specific antigen (PSA) in the diagnosis of prostate cancer. Method: A total of 30 patients with prostate cancer, 30 patients with benign prostatic hyperplasia (BPH) and 30 healthy men (control group) were selected from September 2018 to September 2019, then to detect levels of serum HSP-90α, total PSA and free PSA (FPSA) by ELISA, serum testosterone level by radioimmunoassay, prostate cancer tissue was removed by operation, and relative expression of tissue HSP-90α protein by Western blot. Results: The levels of serum HSP-90α and total PSA in prostate cancer group were significantly higher than other two groups, and testosterone level was lower than other two groups (P < 0.05);there was no difference of serum FPSA level between the three groups (P > 0.05). It was found by Pearson test that serum HSP-90α was positively correlated with total PSA level (r = 0.659, P = 0.005), while negatively correlated with testosterone level (r = -0.549, P = 0.006). According to TNM stage of prostate cancer, there were 17 cases of stage I - II, 13 cases of stage III - IV, 6 cases of Gleason score 1 - 4, 13 cases of 5 - 7, 11 cases of 8 - 10, tumor diameter range from 0.8 to 6.2 cm, with average of (3.9 ± 1.5) cm. The relative expression of HSP-90α protein in tumor tissue was closely related to TNM stage, Gleason score and tumor diameter (P < 0.05). By ROC analysis, it was found that accuracy of combined detection of serum HSP-90α and PSA levels for prostate cancer diagnosis was 0.896, and that of single PSA detection was 0.852. Conclusion: Higher expressions of HSP-90α in prostate cancer tissue and serum may be closely related to occurrence and development of prostate cancer, and combined detections of serum HSP-90α and PSA levels are of great significance in improving early diagnosis of prostate cancer.

热休克蛋白90对hERG-A561V通道蛋白转运及通道功能影响的研究

目的探讨热休克蛋白90(Hsp90)对人类ether-a-go-go相关基因(hERG)-A561V通道蛋白转运及通道功能影响的研究。方法构建野生型(WT)、突变型(A561V)、杂合型(WT/A561V)细胞模型,采用蛋白质印迹法检测各组细胞hERG及Hsp90蛋白表达差异。转染减表达空载(Itf NC)、Hsp90减表达(Hsp90-)、过表达空载(Ovp NC)或Hsp90过表达(Hsp90+)质粒至各细胞模型(即Itf NC组、Hsp90-组、Ovp NC组、Hsp90+组),另设不加载体的对照(Ctl)组,采用蛋白质印迹法检测各组细胞调控Hsp90前后hERG及Hsp90蛋白表达差异。采用免疫荧光法检测Hsp90调控前后杂合型细胞模型hERG及Hsp90蛋白定位表达情况。采用全细胞膜片钳技术检测调控Hsp90前后杂合型细胞模型hERG通道激活电流及尾电流密度。结果突变型、杂合型细胞模型Hsp90及hERG(135 kDa)蛋白表达水平均较野生型明显升高(均P<0.05)。野生型细胞模型Hsp90+组hERG(135 kDa)、hERG(155 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05),突变型细胞模型Hsp90+组hERG(135 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05),杂合型细胞模型Hsp90+组hERG(155 kDa)蛋白表达水平较Ctl组、Ovp NC组均明显升高(均P<0.05)。融合Hsp90和hERG蛋白后,Ctl组hERG表达于细胞膜、细胞质,Hsp90-组细胞膜上hERG蛋白表达减少,Hsp90+组细胞膜上hERG蛋白表达增多。与Ctl组比较,Hsp90+组Ikr曲线左移14.709,斜率因子k值未见明显变化。杂合型细胞模型Ctl组、Hsp90-组、Hsp90+组激活电流及尾电流密度均在50 mV处达到最高;杂合型细胞模型Hsp90+组在20、30 mV处的激活电流密度均明显高于Ctl组、Hsp90-组(均P<0.05),在30、40 mV处的尾电流密度均明显高于Ctl组、Hsp90-组(均P<0.05)。结论Hsp90在hERG-A561V通道蛋白折叠转运中起作用,而上调Hsp90可促进WT/A561V hERG的折叠转运,进而影响通道功能。

柔嫩艾美耳球虫热休克蛋白90和组蛋白4对DF-1细胞凋亡的影响

为了探究柔嫩艾美耳球虫热休克蛋白90(EtHSP90)和组蛋白(EtH4)在体外对鸡胚成纤维细胞(DF-1细胞)凋亡的影响,本试验构建真核荧光表达质粒pEGFP-N1-EtHSP90和pEGFP-N1-EtH4并转染至DF-1细胞,用荧光显微镜观察转染效果,Western blot验证蛋白表达,流式细胞术检测DF-1细胞凋亡。结果显示,经验证成功构建重组质粒pEGFP-N1-EtHSP90和pEGFP-N1-EtH4。流式细胞术检测发现,pEGFP-N1-EtHSP90组早期凋亡率显著低于空白组、pEGFP-N1组和pEGFP-N1-EtH4组(P<0.05);pEGFP-N1-EtH4组早期凋亡率与空白组和pEGFP-N1组相比差异不显著(P>0.05)。因此,EtHSP90早期可抑制细胞凋亡,EtH4既不促进细胞凋亡,也不抑制细胞凋亡。本试验结果为后续深入探究2个蛋白的凋亡机制提供理论基础。

高危型HPV阳性患者血清中HDAC6和Hsp90联合检测在宫颈癌前病变诊断中的临床意义

目的探究高危型人乳头瘤病毒(HR-HPV)阳性患者血清中组蛋白脱乙酰酶6(HDAC6)和热休克蛋白90(Hsp90)联合检测在宫颈癌前病变诊断中的临床意义。方法纳入2020年6月至2021年6月在山东第一医科大学附属济南妇幼保健院接受治疗的166例HR-HPV阳性患者作为研究对象,根据阴道镜检查和活检结果将研究对象分为实验组[高级别鳞状上皮内病变(HSIL)56例、低级别鳞状上皮内病变(LSIL)30例]、对照组(子宫颈良性病变患者80例)。采用酶联免疫吸附试验(ELISA)法检测患者血清HDAC6、Hsp90水平;采用Logistic多因素回归分析HR-HPV患者发生宫颈癌前病变的影响因素;采用受试者工作特征(ROC)曲线评价血清HDAC6、Hsp90对HR-HPV患者发生宫颈癌前病变的诊断价值。结果相较于对照组,实验组吸烟、有盆腔炎病史及性传播疾病感染史患者占比和白细胞、中性粒细胞、白介素-6(IL-6)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)、HDAC6、Hsp90水平显著升高(P<0.05)。Logistic多因素回归分析显示,性传播疾病感染史及IL-6、IL-1β、TNF-α、CRP、HDAC6、Hsp90水平均是HR-HPV阳性患者发生宫颈癌前病变的独立危险因素(P<0.05)。ROC曲线结果显示,血清HDAC6、Hsp90水平及二者联合预测HR-HPV阳性患者发生宫颈癌前病变的曲线下面积(AUC)分别为0.759、0.774、0.870,其中联合预测的AUC显著高于二者单独预测的AUC(Z=2.361、2.042,P<0.05)。HSIL患者IL-6、IL-1β、TNF-α、CRP、HDAC6、Hsp90水平显著高于LSIL患者(P<0.05)。结论血清HDAC6、Hsp90在HR-HPV阳性发生宫颈癌前病变患者中水平升高,均是HR-HPV阳性患者发生宫颈癌前病变的独立危险因素,有望成为HR-HPV阳性患者发生宫颈癌前病变的诊断因子。

血清热休克蛋白70、热休克蛋白90和胃泌素水平对重症颅脑损伤并发应激性溃疡的预测

目的探讨血清热休克蛋白70(HSP70)、热休克蛋白90(HSP90)和胃泌素(GAS)水平对重症颅脑损伤并发应激性溃疡的预测价值。方法选取126例重症颅脑损伤患者,入院后均采用酶联免疫法(ELISA)检测血清HSP70、HSP90和GAS水平,并实施标准去大骨瓣减压术联合腰大池引流术。统计应激性溃疡发生率,分析血清HSP70、HSP90和GAS水平与患者并发应激性溃疡的关系及三者联合对该并发症的预测价值。结果应激性溃疡发生率为32.17%;并发应激性溃疡患者入院后血清HSP70、HSP90和GAS水平均高于未并发者(P<0.05);年龄、入院格拉斯哥昏迷量表(GCS)3~5分、入院后血清HSP70水平、入院后血清HSP90水平、入院后血清GAS水平、入院后转铁蛋白(TRF)水平、入院后随机血糖水平、低血压、低氧血症、代谢性酸中毒、应用糖皮质激素均是并发应激性溃疡的危险因素(P<0.05),入院后血红蛋白(Hb)水平、入院后红细胞压积(HCT)、应用保护胃黏膜药物、应用乌司他丁是其保护因素(P<0.05);血清HSP70、HSP90、GAS水平联合预测重症颅脑损伤并应激性溃疡的灵敏度均高于单独预测(P<0.01),曲线下面积(AUC)均高于单独预测(P<0.05),特异度均与单独预测差异无统计学意义(P>0.05)。结论血清HSP70、HSP90、GAS水平升高可增加重症颅脑损伤并发应激性溃疡的风险,且年龄、入院GCS3~5分等也均是其危险因素,入院后Hb水平、入院后HCT等是其保护因素,血清HSP70、HSP90、GAS水平联合预测并发应激性溃疡的效能高。

多发性骨髓瘤患者血清HSP90 mRNA、β2-MG的表达水平及其与患者预后的关系

目的:探讨多发性骨髓瘤患者血清热休克蛋白90(HSP90)mRNA、β2-微球蛋白(β2-MG)的表达水平及其与患者预后的关系。方法:选取我院94例多发性骨髓瘤患者作为骨髓瘤组,并分为ISS I期26例、ISS II期36例、ISS III期32例;同期选取96例健康体检者作为健康对照组。实时荧光定量PCR(qRT-PCR)法检测血清HSP90 mRNA表达水平,酶联免疫吸附(ELISA)法检测血清β2-MG表达水平。Pearson法分析多发性骨髓瘤患者血清HSP90 mRNA与β2-MG水平的相关性。Kaplan-Meier法分析多发性骨髓瘤患者预后与血清HSP90 mRNA、β2-MG表达水平的关系。多发性骨髓瘤患者预后的影响因素采用Cox回归分析。结果:骨髓瘤组血清HSP90 mRNA、β2-MG水平均明显高于健康对照组(P<0.001)。多发性骨髓瘤ISS III期患者血清HSP90 mRNA、β2-MG水平均明显高于ISS II期和ISS I期患者,ISS II期患者血清HSP90 mRNA、β2-MG水平均明显高于ISS I期患者(P<0.001)。多发性骨髓瘤患者血清HSP90 mRNA与β2-MG水平呈正相关(r=0.630,P<0.001)。HSP90 mRNA高表达多发性骨髓瘤患者3年生存率(35.42%)明显低于HSP90 mRNA低表达患者(63.04%)(P<0.01);β2-MG高表达多发性骨髓瘤患者3年生存率(35.56%)明显低于β2-MG低表达患者(61.22%)(P<0.05)。血清HSP90 mRNA、β2-MG是影响多发性骨髓瘤患者预后的独立危险因素(P<0.05)。结论:HSP90 mRNA、β2-MG在多发性骨髓瘤患者血清中呈高表达,检测二者水平对多发性骨髓瘤患者病情评价及预后评估有潜在参考价值。

青少年鼻咽血管纤维瘤中HSP90表达及临床意义

目的探讨青少年鼻咽血管纤维瘤(JNA)中热休克蛋白90(HSP90)的表达及其与肿瘤复发的关联。方法回顾性分析了2018年—2022年在我院接受手术治疗的组织学诊断为JNA的男性患者60例,使用包含60例JNA患者和10例对照受试者的组织微阵列进行免疫组织化学以评估HSP90表达,通过Pearson卡方、Spearman以及单变量和多变量Cox回归分析HSP90表达与临床病理特征和肿瘤复发的关系。结果与正常中鼻甲样本相比,免疫组织化学显示JNA中HSP90高表达。HSP90高表达与微血管密度(MVD)(R=0.379,P=0.001)、雌激素受体α(ER-α)(R=0.396,P=0.001)、血管内皮生长因子(VEGF)(R=0.612,P<0.001)和JNA复发(P=0.011)呈正相关,是复发时间的独立预后因素(HR=3.251,95%CI:1.187-8.634,P=0.012)。结论HSP90可能是接受手术治疗的JNA患者的独立预后标志物。

莲草直胸跳甲热激蛋白90基因的分子特征、表达模式及对温度胁迫的响应

莲草直胸跳甲Agasicles hygrophila是世界性入侵杂草空心莲子草Alternanthera philoxeroides的专食性天敌,冬季低温和夏季高温会影响其种群数量从而降低防控效果。为进一步探讨莲草直胸跳甲对温度胁迫适应性的分子生态机制,本研究对其热激蛋白90(AhHSP90)进行了蛋白结构、磷酸化修饰及系统发育等生物信息学分析,并采用荧光定量PCR检测了AhHSP 90基因的时空表达动态及不同温度胁迫后的表达。RT-qPCR结果显示,AhHSP 90在莲草直胸跳甲整个生活史及雌成虫各组织中均有表达;高温(30、35和40℃)、低温(-5、0、5、10、15和20℃)胁迫2 h后,AhHSP 90在高温40℃下表达显著升高,但在其他温度下无显著差异。根据对AhHSP 90分子特征、表达模式以及对温度胁迫的响应等方面的综合分析,推测其可能在莲草直胸跳甲发育及抵抗高温过程中发挥着重要的作用。

HSP90通过PTK2B影响Aβ_(1-42)诱导的痴呆小鼠学习记忆功能

目的:探讨热休克蛋白90(HSP90)和蛋白酪氨酸激酶2β(PTK2B)与Aβ_(1-42)诱导痴呆模型小鼠学习记忆功能的关系。方法:32只C57BL/6J小鼠分为对照组(NS)、痴呆模型组(Aβ_(1-42)+NS)、HSP90抑制剂Luminespib处理组(Aβ_(1-42)+Lum)以及PTK2B抑制剂PF431396处理组(Aβ_(1-42)+PF)。实验组侧脑室注射Aβ_(1-42)构建痴呆模型后分别注射HSP90抑制剂或PTK2B抑制剂进行处理。采用水迷宫对小鼠学习记忆功能进行检测,Western Blot检测海马HSP90、PTK2B的表达情况以及tau蛋白磷酸化水平(p-tau),免疫组织化学染色检测海马HSP90和PTK2B的表达,real time RT-PCR检测PTK2B mRNA表达。结果:在Aβ_(1-42)诱导的痴呆模型小鼠海马中,PTK2B及p-tau水平均升高,HSP90水平降低(P<0.01)。而应用HSP90抑制剂则更进一步导致小鼠海马PTK2B、PTK2B mRNA及p-tau的水平升高(P<0.05),并使小鼠学习记忆功能减退;抑制PTK2B后小鼠海马中tau磷酸化水平降低(P<0.01),且小鼠学习记忆功能得到改善,但HSP90表达水平无明显变化(P>0.05)。结论:Aβ_(1-42)诱导的痴呆模型小鼠海马组织中HSP90降低,PTK2B升高。抑制HSP90可能通过促使PTK2B表达和tau蛋白磷酸化水平上升使小鼠学习记忆功能减退。

免疫相关基因NF-κB、IL-17、HSP90在三阴型乳腺癌中的表达及其与淋巴结转移和预后的关系

目的探讨免疫相关基因核因子-κB(NF-κB)、白细胞介素-17(IL-17)、热休克蛋白90(HSP90)在三阴型乳腺癌(TNBC)中的表达及其与淋巴结转移和预后的关系。方法选取2015年10月至2017年12月佛山市中医院诊治的TNBC患者40例作为研究对象,比较TNBC癌组织和癌旁正常组织NF-κB、IL-17、HSP90的表达及其与淋巴结转移和预后的关系。结果TNBC癌组织中NF-κB、IL-17、HSP90的阳性表达率大于癌旁正常组织(P<0.05);Spearman相关分析显示,NF-κB表达与IL-17、HSP90表达呈正相关(P<0.05);IL-17表达与HSP90表达呈正相关(P<0.05)。有淋巴结转移组NF-κB、IL-17、HSP90阳性表达率大于无淋巴结转移组(P<0.05)。二元logistic回归分析结果显示,NF-κB、IL-17、HSP90表达均为TNBC患者淋巴结转移的影响因素(P<0.05)。COX回归分析结果显示,淋巴结转移、远处转移、NF-κB阳性表达、IL-17阳性表达、HSP90阳性表达均为TNBC患者预后的独立影响因素(P<0.05)。结论NF-κB、IL-17、HSP90在TNBC中的阳性表达与淋巴结转移密切相关,可作为预后评估的重要指标。