Autoantibody to heat shock protein Hsp40 in sera of lung cancer patients.

Heat shock protein Hsp40 is a stress protein with chaperone activity and has a cooperative function with Hsp70 in mammalian cells. We examined the possible expression of Hsp40 in lung tumor tissues using immunoblotting and immunohistochemistry, and established an enzyme-linked immunosorbent assay (ELISA) method to detect IgG antibody to Hsp40 in the serum using purified human Hsp40. Sera were obtained from 130 normal subjects and 50 patients with lung cancer. Lung tumor tissues and cells specifically overexpressed Hsp40, and no such expression was detected in normal lung tissues. Compared with normal sera, significantly higher levels of autoantibody to Hsp40 were present in patients with lung cancer. The present study is the first to demonstrate overexpression of Hsp40 in human tumor tissue and the associated presence of autoantibody to Hsp40 in the serum. These results suggest that overexpression of Hsp40 in tumor cells may be recognized as a self-antigen.     

Expression of heat shock protein (Hsp) 70 and Hsp 40 in colorectal cancer

Heat shock protein (Hsp) 70 and Hsp 40 are stress proteins that cooperate as chaperones in mammalian cells. The present study was designed to determine the expression levels of Hsp 70 and Hsp 40 in colorectal cancer by immunohistochemistry and Western blot analysis. Among 50 colorectal cancer tissues studied, 80% and 14% of tumors showed specific immunoreactivity to Hsp 70 and Hsp 40, respectively. Hsp 70 and Hsp 40 were overexpressed in cancer tissue samples compared with normal tissues, on both analytic sets. However, there were no significant correlations between their expression and various clinicopathological parameters of colorectal cancer. Hsp 70 and Hsp 40 may be tumor markers for colorectal cancer.     

Identification and verification of Hsp90-beta as a potential serum biomarker for lung cancer

Background: Hsp90-beta was investigated as prognostic factor because of its apparent association with tumorigenesis. The aim of this study was to investigate the expression of Hsp90-beta in lung cancer patients, to analyze the relationship with respect to the clinicopathological features and to assess whether Hsp90-beta as a potential serum marker for lung cancer. Methods: Expression of Hsp90-beta was examined using immunohistochemistry, in-situ hybridization, western blot and enzyme-linked immunosorbent assay. Sensitivities and specificities for Hsp90-beta serum test were determined using receiver operator characteristic curve and cutoff was defined based on 95% and 85% sensitivities. Results: Lung cancer tissues exhibited higher expression of Hsp90-beta than the normal tissues (P < 0.05) and the serum Hsp90-beta of lung cancer patients also exhibited higher level than control groups (P < 0.05). Moreover, increased serum Hsp90-beta was significantly associated with the pathological grade and clinical stage of lung cancer patients (P < 0.05). Using receiver operator characteristic curve analysis, the cutoffs for distinguishing lung cancer from normal and benign groups were 1.155 and 1.158 ng/ml respectively. The sensitivities of Hsp90-beta for distinguishing lung cancer from normal and benign groups were 98.77% and 95.9%, and specificities were 88.33% and 72.7%. Conclusion: Up-regulation of serum Hsp90-beta was associated with pathological grade and clinical stage of lung cancer patients, which indicated that it could be considered molecular biomarker for diagnosis and prognosis of lung cancer.     

Livin蛋白和STAT3蛋白在支气管肺癌中的表达及临床意义

目的:探讨Livin蛋白和STAT3蛋白在支气管肺癌组织中的表达,及蛋白表达与临床病理特征的关系。方法:采用免疫组化二步法（SP法）检测在支气管肺癌组织及癌旁病理证实的正常肺组织和肺部良性病变组织中Livin蛋白和STAT3蛋白的表达情况。结果:肺癌组的Livin蛋白和STAT3蛋白阳性表达率均高于对照组(均为P<0.05)。男性肺癌患者肺组织、有淋巴结转移肺组织、吸烟指数≥400年支的肺癌患者肺组织中Livin蛋白和STAT3蛋白的阳性表达率均较高,差异有统计学意义(P<0.05)。肺腺癌组织Livin蛋白的阳性表达率显著高于肺鳞癌组织,差异有统计学意义（P=0.009）。低分化肺癌组织中STAT3蛋白阳性表达率明显高于中分化肺癌组织,差异有统计学意义（P=0.004）。支气管肺癌患者的年龄、临床分期对Livin蛋白和STAT3蛋白的阳性表达率的影响均无统计学差异（P>0.05）。Livin蛋白与STAT3蛋白的表达呈正相关。结论:Livin蛋白与STAT3蛋白在支气管肺癌组织中过表达,有望为肿瘤诊断及基因治疗提供新的靶点。Livin蛋白与STAT3蛋白在支气管肺癌组织中表达呈正相关。     

YKL-40表达与子宫内膜癌临床病理因素的关系

目的探讨甲壳质酶蛋白-40（YKL-40）在子宫内膜癌患者的组织和血清中的表达与其临床病理因素的关系。方法采用SP免疫组织化学方法和ELISA方法检测YKL-40在31例子宫内膜癌、10例子宫内膜不典型增生和10例子宫内膜正常者的组织和血清中的表达,并用SPSS 17.0统计软件包分析YKL-40表达与子宫内膜癌的临床病理危险因素的关系。结果①子宫内膜癌患者的YKL-40血清浓度（中位数102.58 ng/ml）明显高于子宫内膜不典型增生（中位数58.20 ng/ml）和正常对照者（中位数37.04 ng/ml）,差异有统计学意义（P〈0.05）;且血清YKL-40表达水平与肿瘤分期呈正相关（P=0.046）,但与组织学分级、淋巴结转移、腹腔冲洗液或腹腔积液细胞学结果无关（P〉0.05）。②YKL-40在子宫内膜癌组织中的阳性表达率为45.2%（14/31）.明显高于子宫内膜不典型增生的10.0%（1/10）和正常对照者的10.0%（1/10）.差异有统计学意义（P〈0.05）,且YKL-40阳性表达与组织学分级呈正相关（P=0.049）,但与临床分期和淋巴结转移无关（P〉0.05）。结论 YKL-40在子宫内膜癌的组织和血清水平均呈高表达,且组织中的阳性表达与肿瘤的组织学分级呈正相关,血清中表达与肿瘤分期呈正相关提示YKL-40可能与子宫内膜癌的发生、发展有关,有望成为子宫内膜癌的诊断和/或预后相关的分子标志物,但尚需进一步验证。     

细胞分裂周期7蛋白在肝癌组织的表达及其对肝癌细胞增殖和侵袭的影响

目的 检测细胞分裂周期7(CDC7)蛋白在人肝癌组织的表达,探讨CDC7过表达对肝癌细胞增殖和侵袭的影响。方法 实时定量聚合酶链式反应(Real-time PCR)检测CDC7 mRNA在肝癌组织和癌旁组织的表达,检测CDC7 mRNA在肝癌细胞系和正常肝脏细胞中的表达。Western blot检测CDC7蛋白在肝癌组织和癌旁组织中的表达,检测CDC7蛋白在肝癌细胞系和正常肝脏细胞中的表达水平。利用慢病毒载体构建稳定过表达CDC7的肝癌HCCLM3和SMMC 7721细胞株。细胞克隆实验和CCK-8法检测CDC7过表达对肝癌细胞增殖的影响。Transwell实验和划痕实验检测CDC7过表达对肝癌细胞侵袭迁移的影响。结果 与癌旁组织相比,CDC7 mRNA和蛋白在肝癌组织中表达水平显著升高(P＜0.001);与正常肝细胞相比,肝癌细胞系的CDC7 mRNA和蛋白表达水平显著增高(P＜0.001);CCK-8法、细胞克隆实验说明CDC7过表达会明显增强肝癌细胞的增殖能力;划痕实验、Transwell实验说明CDC7过表达会明显提高肝癌细胞的侵袭能力。结论 CDC7蛋白在人肝癌组织高表达,其过表达促进肝癌细胞的增殖和侵袭能力。     

人附睾蛋白4在非小细胞肺癌组织及血清中的表达及临床意义

目的 研究人附睾分泌蛋白4(HE4)在非小细胞肺癌(NSCLC)组织及血清中的表达及临床意义.方法 收集124例NSCLC患者(NSCLC组)的血液标本和肺癌组织标本,其中行手术治疗的患者取癌旁组织和正常肺组织,并在术后3天再次抽取血液标本;同期收集96例健康体检者(健康组)的血清标本.采用电化学发光免疫分析仪检测血清中HE4浓度;免疫组化EnVision二步法检测非小细胞肺癌组织、癌旁组织、正常肺组织中HE4的表达.结果 HE4在肺癌组织、肺腺癌组织、Ⅲ~Ⅳ期肺癌组织中的阳性表达率分别高于癌旁组织和正常肺组织、肺鳞癌组织、Ⅰ~Ⅱ期肺癌组织(P<0.05);NSCLC组患者的血清HE4浓度高于健康体检组(P<0.05);手术治疗的患者术后3天血清HE4浓度低于术前血清HE4浓度(P<0.05);NSCLC组患者血清HE4水平与病理分期、组织中HE4阳性表达呈正相关,与年龄、性别、病理类型无明显相关性.结论 NSCLC组织及血清HE4表达升高,可作为NSCLC的肿瘤标志物,是NSLCL的早期辅助诊断、疗效评价、预后评估的客观指标,可用于指导临床的综合治疗.     

非小细胞肺癌中WWOX蛋白及ErbB2蛋白的联合检测

 目的 探讨WWOX(WW domain-containing oxidoreductase)基因在非小细胞肺癌组织中的表达及与ErbB2蛋白之间的关系。 方法 用免疫组化方法检测50例肺鳞癌,31例肺腺癌及20例癌旁正常组织中WWOX基因蛋白的表达,并分析其与各临床病理指标及ErbB2蛋白之间的相关性。 结果 WWOX基因在72.8%的非小细胞肺癌中失表达或表达减少,而在癌旁正常组织中有80.0%正常表达。WWOX基因的表达与组织类型、细胞分化程度密切相关（P<0.05）,在鳞癌和低分化癌中WWOX基因失表达或表达减少的发生率更高。肺癌中ErbB2的过度表达与肺癌的临床分期、淋巴结转移相关,WWOX蛋白阳性表达与ErbB2负相关(r=–0.239,Ｐ<0.05)。 结论 WWOX蛋白的低表达与ErbB2的过表达可以协同判断预后较差的非小细胞肺癌。     

EphA2蛋白表达与结直肠癌浸润性及微血管生成的关系

目的：探讨酪氨酸蛋白激酶受体EphA2表达与大肠癌浸润及其与大肠癌微血管生成之间的关系。方法：应用免疫组化方法检测70例大肠癌标本和相应正常大肠黏膜组织中EphA2蛋白的表达情况,分析E-phA2表达与大肠癌临床病理学因素及微血管生成之间的关系。结果：EphA2蛋白在癌组织中的表达明显高于相应正常大肠黏膜组织（P〈0.001）。EphA2蛋白高表达与癌分化程度、生长方式、浸润深度和肿瘤体积相关（P〈0.05）,而与患者的年龄、性别、淋巴结和血行性转移无关（P〉0.05）。免疫组化结果还发现,肿瘤组织内微血管内皮细胞也有EphA2受体蛋白表达。CD34染色后大肠癌的微血管密度（microvessel density,MVD）与EphA2表达水平有显著相关,EphA2阳性表达强度高的肿瘤区域有较高的微血管密度。结论：E-phA2蛋白的高表达与大肠癌的发生和浸润有关。     

非小细胞肺癌移植瘤侵袭组织中血清蛋白的分布

目的：局部侵袭是肺癌患者死亡的主要原因之一,本研究探讨肺癌侵袭组织中血清蛋白的分布及其与血管内皮生长因子（VEGF）和表皮生长因子受体（EGFR）蛋白的关系。方法：建立非小细胞肺癌（NSCLC） A549细胞注射裸鼠背部皮下的异种移植模型,利用活体冷冻技术（IVCT）制备标本,免疫组化分析Albumin、IgG、IgM、VEGF、EGFR蛋白分布。结果：观察到NSCLC侵袭组织周围结缔组织、血管、间质及细胞外基质有Albumin和IgG1、IgM主要在侵袭组织周围结缔组织和血管内,癌细胞外基质未观察到IgM。肺癌细胞浆内VEGF蛋白呈细颗粒状,但是癌细胞浆和膜未观察到EGFR蛋白分布。结论：利用IVCT技术能清晰观察到侵袭组织周围结缔组织、血管和细胞外基质中不同分子量血清蛋白分布,NSCLC侵袭组织细胞外基质Albumin、IgG1和IgM免疫组织化学染色分布主要依据其分子量,与VEGF和EGFR蛋白分布无关。     

人肺癌组织细胞Na+/H+交换蛋白-1基因的表达与其抗凋亡的关系

目的 :通过对临床人肺癌组织Na /H 交换蛋白 1(NHE 1)mRNA的表达的检测 ,探讨肿瘤治疗新的特异靶点。方法 :采用RNA原位分子杂交法 (RNA IMH)检测了 2 7例人肺癌组织和正常肺组织NHE 1mRNA的表达。结果 :人肺癌组织细胞NHE 1mRNA的表达显著增强 ,呈过表达 ,而各病理类型间无显著差异。结论 :人肺癌组织细胞中NHE 1mRNA过表达这一区别于正常组织细胞的分子生物学特征与肿瘤细胞的抗凋亡特性的形成密切相关 ,可能为肿瘤治疗学提供良好的特异靶点并具有重要的潜在临床意义     

Heat shock protein 70 (Hsp70) membrane expression on head-and-neck cancer biopsy-a target for natural killer (NK) cells

PURPOSE: Heat shock protein 70 (Hsp70) was detected on the cell membrane of human tumor cell lines, but not on normal cells. Here we studied Hsp70 membrane expression as a target for natural killer (NK) cells on tumor material and control tissues of head-and-neck cancer patients. METHODS AND MATERIALS: Membrane-bound Hsp70 was determined by flow cytometry on single-cell suspensions of tumors and the corresponding normal tissues of head-and-neck cancer patients. The cytolytic activity of NK cells against Hsp70-positive tumor cells was measured in a standard cytotoxicity assay. RESULTS: In total, 54 of 74 primary tumors were found to be Hsp70 membrane-positive (73%); tongue/mouth, 21 of 24 (88%); oropharynx, 13 of 20 (65%); hypopharynx, 3 of 6 (50%); larynx, 8 of 11 (73%); trachea 1 of 2 (50%); esophagus, 4 of 5 (80%); lymph node metastases, 4 of 6 (67%). The corresponding control tissue was negative for membrane-bound Hsp70. Biopsies (6 of 6) of patients after in vivo gamma-irradiation (fractionated 5 x 2 Gy) were strongly Hsp70 membrane-positive. Irradiated, Hsp70-positive tumor cells are targets for Hsp70-peptide stimulated NK cells. CONCLUSION: An irradiation-inducible, tumor-selective Hsp70 membrane localization provides a target structure for Hsp70-peptide stimulated human NK cells.     

非小细胞肺癌血清标志物PACAP的蛋白质组学筛选和鉴定

背景与目的早诊断早治疗可以提高肺癌的长期生存率,血清标志物可能有助于肺癌的早期诊断。本研究在组织培养的基础上,应用蛋白质组学方法筛选潜在的肺癌血清标志物。方法采用差异蛋白质组学技术对肺腺癌和肺组织培养液中的差异蛋白质进行初步鉴定。选取其中的半胱天冬酶凋亡前衔接蛋白(proapoptotic caspaseadapter protein,PACAP)做更进一步的研究,用Western blot验证PACAP在培养液上清中的表达情况并检测它在不同类型血清中的表达情况,用ELISA检测不同类型血清标本中PACAP的表达情况。结果共得到差异蛋白点19个,鉴定出14个。PACAP在双向电泳中表达差异较大,Western blot显示PACAP在肺癌组织培养液及肺癌患者血清中的表达均明显增高。ELISA显示PACAP在肺腺癌和肺鳞癌患者血清中的表达水平高于在小细胞肺癌、肺良性肿瘤和健康人血清中的水平(P<0.05)。结论 PACAP是潜在的非小细胞肺癌血清标志物。     

Hsp90 is expressed and represents a therapeutic target in human oesophageal cancer using the inhibitor 17-allylamino-17-demethoxygeldanamycin

Heat shock protein 90 (Hsp90) has been demonstrated to protect oncogenic variants of signalling molecules from degradation and may consequently serve as a therapeutic target for the treatment of oesophageal cancer for which adequate therapy is often lacking. We studied the expression of Hsp90 in tumour tissues of human oesophageal cancer and the impact of Hsp90 inhibition on oesophageal cancer cell lines using the drug 17-allylamino-17-demethoxygeldanamycin (17-AAG). Quantitative immunohistochemistry was performed on formalin-fixed paraffin-embedded tissues from patients with oesophageal cancer. In squamous cell carcinoma, a marked upregulation of Hsp90 could be noted in dysplastic epithelium and invasive cancer compared with normal epithelium. In adenocarcinoma, Hsp90 was expressed in neoplastic epithelium and also in normal non-neoplastic glands weakly. The inhibition of Hsp90 using 17-AAG led to a significant decrease in cell proliferation and viability in human oesophageal cancer cell lines. Using a clonogenic cell survival assay, Hsp90 inhibition significantly sensitised the cells for γ-photon irradiation. Heat shock protein 90 was found to be critical for proper signalling induced by both epidermal growth factor and insulin-like growth factor-1, in which the inhibition of signalling by 17-AAG correlated with the observed reduction in cell proliferation and viability. These results showed that Hsp90 was selectively expressed in oesophageal cancer tissue compared with the corresponding normal tissue, and the inhibition of Hsp90 resulted in decreased proliferation and viability as well as radiosensitisation of oesophageal cancer cells. Heat shock protein 90 represents a potential therapeutic target in the treatment of patients with oesophageal cancer, alone or in combination with radiotherapy.     

Targeted Efficacy of Dihydroartemisinin for Translationally Controlled Protein Expression in a Lung Cancer Model

Objective: Lung cancer is one of the malignant tumors with greatest morbidity and mortality around theworld. The keys to targeted therapy are discovery of lung cancer biomarkers to facilitate improvement of survivaland quality of life for the patients with lung cancer. Translationally controlled tumor protein (TCTP) is one ofthe most overexpressed proteins in human lung cancer cells by comparison to the normal cells, suggesting thatit might be a good biomarker for lung cancer. Materials and Methods: In the present study, the targeted efficacyof dihydroartemisinin (DHA) on TCTP expression in the A549 lung cancer cell model was explored. Resultsand Conclusions: DHA could inhibit A549 lung cancer cell proliferation, and simultaneously up-regulate theexpression of TCTP mRNA, but down-regulate its protein expression in A549 cells. In addition, it promotedTCTP protein secretion. Therefore, TCTP might be used as a potential biomarker and therapeutic target fornon-small cell lung cancers.     

DEK is highly expressed in breast cancer and is associated with malignant phenotype and progression

DEK proto-oncogene (DEK) has been demonstrated as an oncogene and is associated with the development of many types of tumor; however, the expression and role of DEK in breast cancer remain unknown. The present study aimed to determine the role of DEK in the progression of breast cancer. The expression of DEK in 110 breast cancer tissues and 50 adjacent normal breast tissues was examined using immunohistochemistry. Furthermore, DEK expression was upregulated by DEK transfection or downregulated by DEK shRNA interference in MCF7 cells. Proliferative and invasive abilities were examined in MCF7 cells using MTT assay, colony-formation assay and transwell invasion assays. The results demonstrated that DEK expression level was significantly increased in breast cancer tissues compared with normal breast tissues. Furthermore, high DEK expression was associated with high histological grade, lymph node metastasis, advanced Tumor-Node-Metastasis stage and high Ki-67 index; however, DEK expression was not associated with the expression level of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. High DEK expression indicated poor prognosis in patients with breast cancer. DEK overexpression upregulated the protein expression of β-catenin and Wnt and increased the proliferative and invasive abilities of breast cancer cells. DEK downregulation had the opposite effect. Taken together, the results from the present study demonstrated that high expression of DEK was common in patients with breast cancer and was associated with progression of the disease and poor prognosis, and that DEK overexpression promoted the proliferative and invasive abilities of breast cancer cells.     

linc00675在肺癌组织和细胞中的表达及其抑制肺癌细胞迁移和侵袭的机制

目的:探讨linc00675在肺癌组织和细胞中的表达以及其对肺癌细胞迁移、侵袭的影响。方法:采用qRT-PCR检测linc00675在35例肺癌组织和癌旁组织及肺癌细胞株SPCA1、A549、NCI-H446和人肺正常上皮细胞BEAS-2B中的表达。利用细胞转染实验对肺癌细胞SPCA1进行LV-linc00675和siRNA-linc00675及相关阴性对照的转染,采用qRT-PCR检测linc00675的表达水平。采用Transwell实验检测过表达及干扰linc00675表达对SPCA1细胞迁移和侵袭能力的影响。通过Western blot实验检测细胞上皮标志蛋白 E-钙黏蛋白（E-cadherin）、间质标志蛋白 N-钙黏蛋白（N-cadherin）、波形蛋白（Vimentin）的表达水平。结果:linc00675在肺癌组织中的表达显著低于癌旁组织,在肺癌细胞株中的表达显著低于人肺正常上皮细胞。转染LV-linc00675可显著增加linc00675在肺癌细胞中的表达,转染siRNA-linc00675可显著降低linc00675在肺癌细胞中的表达。Transwell实验结果显示,过表达linc00675可显著抑制肺癌细胞的迁移和侵袭能力,干扰linc00675表达可显著增强肺癌细胞的迁移和侵袭能力。Western blot实验结果显示,过表达linc00675可显著抑制SPCA1细胞上皮间质转化（EMT）的发生,干扰linc00675表达后SPCA1细胞发生了明显的EMT。结论:linc00675在肺癌细胞中发挥抑癌作用,linc00675可能通过抑制肺癌细胞EMT的发生从而减弱其迁移和侵袭能力,提示linc00675可能成为肺癌治疗的新靶点。     

PRR11蛋白在人胰腺癌中的表达及其意义

背景与目的：近期研究发现,新基因PRR11在肺癌和胃癌中表达异常,推测可能与肿瘤进展有关。本研究探讨PRR11蛋白在人胰腺癌中的表达,并探讨其与胰腺癌临床病理参数之间的关系。方法：采用免疫组织化学SP法检测32例人胰腺癌组织、20例癌旁组织和6例正常胰腺组织中的PRR11蛋白的表达,并采用χ2检验分析PRR11蛋白的表达水平与临床病理参数(年龄、性别、肿瘤大小、肿瘤部位、分化程度、淋巴结转移和TNM分期)之间的关系。结果：PRR11蛋白在胰腺癌组、癌旁组织组和正常胰腺组织组的阳性表达率分别为78.1%(25/32)、5.0%(1/20)和0.0%(0/6),胰腺癌组的表达显著高于癌旁组织组及正常胰腺组织组(P<0.05),其在胰腺癌组织中的表达情况与分化程度、TNM分期有关(P<0.05),但与年龄、性别、肿瘤大小、肿瘤部位、有无淋巴结转移无关(P>0.05)。生存分析显示,PRR11蛋白阳性组患者的生存率低于阴性组(P<0.05)。结论：检测PRR11蛋白在胰腺癌中的表达,有望成为判断胰腺癌预后的一个指标。     

肺腺癌患者癌组织中MAP4的表达水平及其与病理参数、预后的关系

目的:分析肺腺癌患者癌组织中微管关联蛋白4(MAP4)的表达水平及其与病理参数、预后的关系.方法:选取我院2014年1月至2015年1月82例非小细胞肺癌(NSCLC)患者,利用免疫组织化学染色法对手术切除的癌组织及其癌旁正常组织的MAP4蛋白表达水平进行检测,并分析MAP4蛋白表达与患者临床病理参数和预后的关系.结果...