Cytoplasmic expression of c-erbB2 in non-small cell lung cancers

The aim of this study was to investigate the expression of c-erbB2 in non-small cell lung cancers (NSCLCs) with attention both to membranous and cytoplasmic reaction, and to try to elucidate the meaning of cytoplasmic expression of c-erbB2 in NSCLCs. Immunohistochemical c-erbB2 expression and related clinico-pathological features were examined in 312 surgically resected patient tissues of NSCLCs, including 175 cases of adenocarcinoma and 137 cases of squamous cell carcinoma. Immunostaining of inner- and ecto-domain of c-erbB2, mRNA expression and the quantitation of soluble c-erbB2 in cultured media were performed in five NSCLC cell lines. Cytoplasmic expression of c-erbB2 was observed more frequently than membranous, both in patient tissues and cell lines. Neither membranous nor cytoplasmic expression of c-erbB2 was significantly correlated with short outcome in NSCLCs. Membranous c-erbB2 was expressed by both inner and ecto-domain, while cytoplasmic c-erbB2 was expressed by either or both inner and ecto-domain. c-erbB2 mRNA was produced in most cell lines; however, the soluble form was only detectable in a cell line that only presented a membranous c-erbB2. In conclusion, cytoplasmic c-erbB2 of NSCLCs was not a full-length protein only expressed in cellular membrane, but reflected degenerated c-erbB2 fragments with less functional ability.     

B7-H6 expression in non-small cell lung cancers

B7 family has been known to be a negative regulator of immunity response in patients with lung cancer. B7-H6 as a novel identified member of B7 family is found to trigger natural killer (NK) cell cytotoxicity and cytokine secretion by binding natural cytotoxicity receptor NKp30. Up until now, no investigations have been made about B7-H6 expression in lung cancer. We present the result of the B7-H6 prognostic value in 65 non-small cell lung cancer (NSCLC) tissues and 65 matched adjacent non-tumor tissues by Immunohistochemistry (IHC). Meanwhile, fluorescence activated cell sorter (FACS) analysis was used to detect B7-H6 receptor NKp30 expression in 7 non-small cell lung cancer tissues and 7 adjacent non-tumor tissues. Here, the result showed B7-H6 immunoreactivity in 6/65 (9.23%) lung cancer patients and 4/65 (6.15%) in adjacent non-tumor tissues. No relationship was found between B7-H6 expression and clinic pathological features. Similarly, no relevance was found for NKp30 expression in lung cancer tissues and non-tumor tissues. However, B7-H6 positive carcinomas were significantly correlated with degree of differentiation (P = 0.044). Three year survival rate after operation did not show the prognostic value for B7-H6 expression. Our study suggests that B7-H6 has a limited value as a prognostic marker in the patients of lung cancer.     

非小细胞肺癌中窖蛋白1基因的蛋白表达和甲基化及其意义

目的 检测窖蛋白1(Car-1)在非小细胞肺癌(NSCLC)中的蛋白表达以及启动子的甲基化状况,探讨Cav-1基因在NSCLC中的作用及其临床意义.方法 应用免疫组织化学(sP法)和量子点Qd600染色检测123例NSCLC组织、17例良性病变肺组织中Cav一1蛋白表达和亚细胞定位.亚硫酸氢钠处理DNA,甲基化特异性PCR(MSP)检测Cav-1基因启动子区域的甲基化水平.结果 Cav-1蛋白在肺支气管黏膜上皮细胞、肺泡上皮细胞、毛细血管内皮细胞、成纤维细胞、平滑肌细胞的胞质和胞膜高表达.癌旁组织(对照)组和肺癌组中Cav-1蛋白的阳性率分别为17/17、43.1%(53/123),两组间差异有统计学意义(P=0.001);Cav-1蛋白在NSCLC不同的组织学类型(P=0.552)和分化程度(P=0.160)中差异均无统计学意义.Cav-1蛋白阳性率与NSCLC的TNM分期(P=0.001)以及淋巴结转移(P=0.001)均相关.在40例Cav-1蛋白表达为阴性的肺癌组织和12例癌旁肺组织,MSP法均未检测到Cav-1因启动子区域的甲基化.结论 Cav-1蛋白失表达的机制可能与启动子区是否甲基化无关.Cav-1蛋白高表达预示NSCLC恶化进展和高侵袭性.     

Differential expression of matrix metalloproteinases and their inhibitors in non-small cell lung cancer

In a comprehensive immunohistochemical study of the expression of ten metalloproteinases (MMPs) and their four inhibitors (TIMPs) in 115 non-small cell lung carcinomas (NSCLCs), the findings have been correlated with the histological and clinical features of the tumours. All MMPs and TIMPs were expressed in tumours, with frequencies ranging from 41% for MMP-2 to 68% for MMP-13. Stromal immunoreactivity ranged from 6% for TIMP-4 to 87% for MMP-13. In some tumours, an overexpression of these proteins, as revealed by stronger staining in cancer cells than in adjacent normal bronchial epithelium, was also observed. The frequency ranged from 1% for MMP-3 to 28% for MMP-13. Compared with squamous cell carcinoma (SqCC), adenocarcinoma (AdC) more frequently overexpressed MMP-1, -11, -13, -14, and TIMP-2, and TIMP-1 and/or TIMP-2 overexpression positively correlated with more advanced stage disease. None of the MMP or TIMP expression correlated with the ras genotype of the tumours. The higher frequency of MMP overexpression in AdC than in SqCC may relate to the greater tendency of the former for systemic metastasis. The association of TIMP-1 overexpression with more advanced disease may suggest a role in prognosis.     

小细胞和非小细胞肺癌晚期患者NK细胞表达的差异

目的:探索小细胞和非小细胞肺癌晚期患者NK细胞是否存在差异,并为治疗提供参考。方法:选取肺癌晚期患者共65例,其中包括小细胞肺癌14例,非小细胞肺癌51例以及20例健康对照。用流式细胞仪检测研究对象外周血淋巴细胞表面CD3-CD16+CD56+的表达情况。结果:小细胞肺癌晚期的患者较健康对照NK细胞显著升高;非小细胞肺癌晚期的患者较健康对照NK细胞无显著变化;肺癌晚期患者外周血NK细胞表达的百分比与CD4+T细胞表达的百分比呈负相关性。结论:小细胞肺癌晚期患者NK细胞升高,天然免疫可能成为已严重受损的细胞免疫的有力补充,但有待于进一步的研究。     

Differential expression of DUSP6 with expression of ERK and Ki-67 in non-small cell lung carcinoma

Dual specificity phosphatase 6 (DUSP6) is a member of the MAP kinase phophatase family. DUSP6 inactivates extracellular signal-regulated kinase (ERK), belonging to the MAP kinase family, and can act in tumor suppressive pathways. The aim of this study was to investigate associations of DUSP6 expression with expression of ERK and Ki-67 and with clinicopathological parameters in lung adenocarcinoma and squamous cell carcinoma. A total of 102 squamous cell carcinomas and 66 adenocarcinomas were studied using immunohistochemistry for DUSP6, ERK1/2, and Ki-67. In 66 adenocarcinomas, high DUSP6 expression was positively correlated with ERK1/2 expression. High DUSP6 expression was correlated with lower histological grade and lower Ki-67 index in the adenocarcinomas. In 102 squamous cell carcinomas, high DUSP6 expression was correlated with lower ERK expression, with greater smoking pack-years, but not with the Ki-67 index. These results indicate that DUSP6 acts as a negative feedback regulator of ERK in adenocarcinoma progression, but that DUSP6 does not play a role in the downregulation of ERK in squamous cell carcinoma. The differential expression of DUSP6 correlated with Ki-67 index, suggesting that DUSP6 plays an important role in cancer resistance in different subtypes of non-small cell lung carcinoma.     

Stathmin在非小细胞肺癌中表达增强* 基金项目：辽宁省科学技术计划立项课题资助（编号：2007225005-13）

 摘要:目的 探讨Stathmin在非小细胞肺癌（non-small cell lung cancer, NSCLC）组织中和正常组织中的表达,了解其与NSCLC临床病理特征之间的关系。方法 用免疫组化与RT-PCR方法,检测43例NSCLC 术后癌组织及正常组织标本中Stathmin蛋白与mRNA的表达。结果 Stathmin蛋白和基因在NSCLC中阳性表达率分别为62.79%和67.44%,显著高于正常组织的16.28%和20.93%（P＜0.01）,其表达与肿瘤细胞分化程度和有无淋巴结转移有关（P ＜0.05）。结论 Stathmin在NSCLC的发生发展过程中起了重要作用,可能成为预测NSCLC恶性程度的新的生物学及个体化治疗的敏感指标。     

Differential expression and distribution of epithelial adhesion molecules in non-small cell lung cancer and normal bronchus

BACKGROUND: Changes in epithelial cell interactions have been implicated in carcinogenesis, tumour invasion and metastasis. AIM: To screen for altered expression of epithelial adhesion genes in lung cancer development. METHODS: Gene expression profiles were assessed with cDNA expression arrays in eight non-small cell lung cancer (NSCLC) and eight normal bronchi obtained from the same patient. Immunohistochemistry (IHC) and RNA in situ hybridisation (ISH) were used to confirm the most prominently expressed adhesion molecules and to investigate their distribution at protein and mRNA levels. RESULTS: 43 differentially expressed cancer-related genes were identified in adenocarcinoma, squamous cell carcinoma (SCC) and normal bronchus. Five of these genes are related to epithelial adhesion-that is, integrin alpha3 (ITGA3), integrin beta4 (ITGB4), desmoplakin I and II (DSP), plakoglobin, and desmocollin 3 (DSC3). ITGA3 and ITGB4, showing predominantly cell-matrix staining, were up regulated in adenocarcinoma and SCC, respectively. ITGB4 also showed strong staining in SCC with IHC and ISH. Components of the desmosome adhesion complex DSP, plakoglobin and DSC3 were strongly up regulated in SCC and showed a distinct cell-cell staining pattern. DSP and plakoglobin were predominantly present at central, more differentiated tumour cells, whereas DSC3 showed a stronger staining in the peripheral basal cells of SCC tumour areas. CONCLUSIONS: Lack of cellular adhesion may have an important role in the metastatic potency of a primary tumour. A possible association of strong presence and normal-distributed desmosomal molecules in SCC with the less frequent and late pattern of metastasis in SCC as compared with adenocarcinoma is suggested.     

Differential expression of heat shock protein 90 isoforms in small cell lung cancer

Heat shock protein 90 (HSP90), a molecular chaperone, plays important roles in cellular protection against various stressful stimuli and in the regulation of cellular growth and apoptosis. HSP90 has 4 different types of human isoforms; HSP90α, HSP90β, glucose related protein 94 (GRP94) and tumor necrosis factor (TNF) receptor-associated protein 1 (TRAP1). We assessed the differential expression of these HSP90 isoforms in small-cell lung cancer (SCLC) and the correlation of their expression levels with clinicopathological factors and patient survival rates. This study included 117 SCLCs, comprised of 108 primary and 9 metastatic tumor tissues. We performed immunohistochemical staining for HSP90α, HSP90β, GRP94 and TRAP1 in 117 tumors and found that HSP90α and HSP90β were positive in 11 (9%) and 61 tumors (52%), respectively, most of which showed weak expression, whereas GRP94 and TRAP1 were positive in 115 (98%) and 117 tumors (100%), respectively, the majority of which showed moderate or strong expression. None of the HSP90 isoforms showed significant associations with clinicopathological factors or survival status in patients with SCLC. Our results indicate that GRP94 and TRAP1 might contribute more to the carcinogenesis or biology of SCLC than HSP90α and HSP90β, and that isoform selectivity should be considered when HSP90 inhibitors are studied or utilized for the treatment of SCLC.     

表达谱芯片筛查非小细胞肺癌细胞系A549放疗抵抗差异基因

目的 建立非小细胞肺癌（NSCLC）A549放疗抵抗模型,表达谱芯片技术筛查放疗抵抗差异基因。 方法 A549细胞系长期间歇照射诱导建立的放疗抵抗细胞模型;表达谱基因芯片筛查放疗抵抗细胞株与敏感株全基因组RNA表达差异;分析2倍以上差异的基因（P<0.05）,分别进行生物学信息基因分类(GO)和信息通路（Pathway）分析。 结果 表达谱芯片显示,抵抗株与敏感株相比,差异表达基因为1410个（733个上调,677个下调）;GO分析显示,主要与细胞周期、DNA修复有关;通路显著性富集分析显示,丝裂原活化蛋白激酶（MAPK）和磷脂酰肌醇-3-激酶（PI3K）等多条信号通路激酶出现显著性差异。 结论 多种基因和信号通路参与了放疗抵抗过程。     

Histopathologic characteristics of advanced-stage ROS1-rearranged non-small cell lung cancers

Background: ROS1 rearrangement accounts for 1%–2% of non-small cell lung cancer (NSCLC) with a remarkable response to crizotinib. Although ROS1-rearranged tumors are known to have characteristic histologic features, only a few studies have investigated the histologic features of advanced-stage ROS1-rearranged tumors.Methods: We analyzed the histopathologic features of ROS1-rearranged tumors in advanced-stage NSCLC patients and assessed the ROS1 immunohistochemistry (IHC) staining patterns of ROS1-rearranged cases.Results: A total of 37 ROS1 fluorescence in situ hybridization (FISH)-positive cases and 64 ROS1 FISH-negative cases were analyzed, and all tumors were EGFR-, ALK-, and RET-negative. Solid pattern, round nuclei with macronucleoli, solid signet-ring cells, extracellular mucin, and a close relation with adjacent bronchioles were significantly associated with ROS1 rearrangement, and the solid signet-ring cell feature was exclusively identified in ROS1-rearranged tumors. ROS1 IHC showed a 97.3% sensitivity when weak to strong protein expression was considered positive.Conclusions: Our findings highlight distinct histologic features of ROS1-rearranged tumors, including their nuclear features. A thorough understanding of ROS1 rearrangement-related histologic features would be helpful to identify ROS1-rearranged tumors in advanced-stage NSCLC.     

Stathmin在非小细胞肺癌中表达增强

目的探讨stathmin在非小细胞肺癌(NSCLC)组织中和正常肺组织中的表达,了解其与NSCLC临床病理特征之间的关系。方法用免疫组化与RT-PCR方法,检测43例NSCLC术后癌组织及正常肺组织标本中stathmin蛋白与基因的表达。结果 Stathmin蛋白和基因在NSCLC中阳性表达率分别为62.79%和67.44%,显著高于正常肺组织的16.28%和20.93%(P<0.01),其表达与肿瘤细胞分化程度和有无淋巴结转移有关(P<0.05)。结论 Stathmin在NSCLC的发生发展过程中起了重要作用,可能成为预测NSCLC恶性程度的新的生物学及个体化治疗的敏感指标。     

MicroRNA-153 expression and prognosis in non-small cell lung cancer

Background: miR-153 has been found to be significantly decreased in non-small cell lung cancer (NSCLC) tissues; however, its clinical significance has not been investigated. Methods: The expression patterns of miR-153 in 137 pairs of human lung cancer tissues and adjacent normal lung tissues were analyzed using qRT-PCR. The relationships between miR-153 expression and clinicopathological parameters were examined by chi-square test. Kaplan-Meier method and the log-rank test were used to determine the difference in overall survival (OS) rates between two groups. Results: The expression of miR-153 was reduced significantly, compared with adjacent normal lung tissues (P<0.05). We observed that the expression level of miR-153 was positively correlated with the clinical stage (P=0.005), lymph node status (P=0.014), distant metastasis (P=0.004), and differentiated degree (P<0.001) in NSCLC patients. According to the Kaplan-Meier survival analysis, the patients with low miR-153 expression exhibited evidently poorer overall survival rates than those with high miR-153 expression (P=0.003). Multivariate analysis showed that the expression of miR-153 was an independent and significant factor associated with poor OS rates (P=0.002). Conclusion: Decreased expression of miR-153 might be a potential unfavorable prognostic factor for patients with NSCLC, and further studies would be needed to prove our findings.     

SEL1L expression in non-small cell lung cancer

SEL1L gene product plays a role in cell transformation and tumor progression in human breast, pancreas, esophageal, and prostate cancer. SEL1L expression was evaluated in a series of 76 surgically resected non-small cell lung carcinomas to investigate its clinical significance. SEL1L is scarcely detectable in normal lung, whereas in the initial stages of cell transformation, it becomes consistently expressed with evident staining in bronchial squamous metaplasia and in associated dysplastic changes. SEL1L immunoreactivity can be detected both in the cytoplasm and less commonly in the nuclei; the subcellular location correlates with tumor histotype, with cytoplasmic immunoreactivity being most prevalent in squamous cell carcinomas (P = .0005) and nuclear immunoreactivity being associated with adenocarcinomas (P = .02). Nuclear import and export signals are present in the SEL1L coding sequence, justifying the different subcellular location of the protein. SEL1L immunoreactivity was inversely correlated with tumor grade (P = .05); when considering only the adenocarcinomas, a stronger association was found (P = .006). SEL1L messenger RNA and protein evaluation in lung cancer cell lines confirmed the expression of the gene and the dual subcellular location of the protein in lung tumors. The data here reported suggest that, in non-small cell lung carcinoma, SEL1L may be an indicator of cell transformation, thus having important biologic and clinical implications.     

Survivin expression in pre-invasive lesions and non-small cell lung carcinoma

Survivin is an inhibitor of apoptosis protein, which is overexpressed in many carcinomas, including lung carcinoma. The aim of this immunohistochemical study was to investigate the role of survivin in the early steps of lung carcinogenesis and non-small cell lung carcinomas (NSCLC), and its relationship with expression of p53 protein, a tumor suppressor gene involved in cell cycle control. In the normal bronchial epithelium, low-grade atypical adenomatous hyperplasia (AAH) and non-neoplastic lung parenchyma adjacent to tumor, survivin was found completely negative. Expression of survivin was detected in the areas of squamous metaplasia and dysplasia as well as high-grade AAH lesions adjacent to tumor. Survivin was expressed in 50 (64%) and p53 in 41 (53%) NSCLC. Survivin expression was significantly correlated with lymph node metastasis (p=0.02). There was no correlation between survivin and p53 expression. The patients with expression of survivin had significantly worse prognosis (Log-rank test, p=0.003). Multivariate Cox regression analysis showed TNM stage (p<0.001) and survivin expression (p=0.003) as independent prognostic indicators. In conclusion, survivin expression might be an early step in lung carcinogenesis. Survivin expression might also be used as a prognostic indicator predicting the worse outcome in NSCLC, and might be a novel target for the treatment of patients with preinvasive lesions of lung and NSCLC.