肾癌浸润NK细胞活化性受体表达及临床意义

目的:研究肾癌及癌旁组织中NK细胞表面活化性受体NKp44、NKp46表达情况,探讨活化性受体在肾癌发生发展中的临床意义.方法:流式细胞术及免疫组织化学法检测28例肾癌及癌旁组织中NK细胞含量及活化性受体NKp44、NKp46的表达,并结合临床相关因素进行统计学分析.结果:肾癌组织浸润的NK细胞含量明显高于癌旁组织(P＜0.05).肾癌与癌旁组织中活化性受体NKp44表达差异性明显.肾癌病理分级越高NK细胞含量越低,活化性受体NKp44表达也降低.而肾癌与癌旁组织中活化性受体NKp46的表达无统计学意义.结论:肾癌组织中有大量NK细胞浸润,NK细胞活化性受体NKp44表达很低,且NK细胞的含量及活化性受体NKp44的表达是与病理分级相关的.肾癌的发病可能与NK细胞减低及活化性受体NKp44表达降低有关.     

Allelotype and replication error phenotype of small cell lung carcinoma

Allelotype and replication error (RER) phenotype analyses were performed to clarify the pathogenetic significance of inactivation of tumor suppressor genes and genomic instability in the genesis and progression of small cell lung carcinoma (SCLC). We examined 37 cases of SCLC for loss of heterozygosity (LOH) and microsatellite instability at 49 loci on all 39 nonacrocentric chromosomal arms. LOH was frequently (>70%) detected on chromosomes 3p (29/32, 90.6%), 5q (15/21, 71.4%), 13q (25/26, 96.2%), 17p (22/25, 88.0%), and 22q (24/33, 72.7%). Frequent LOH (>70%) on these loci was observed even among seven cases of stage I tumors. The incidence of LOH on all 39 nonacrocentric chromosomal arms was not significantly different between primary tumors and metastases. These results suggest that inactivation of multiple tumor suppressor genes accumulates relatively early during progression of SCLC and it may be responsible for clinically and biologically aggressive phenotype of SCLC. RER was observed in 6/37 (16.2%) of SCLC, however, RER at multiple loci was observed only in two cases. Therefore, it was indicated that genomic instability is uncommon, but might play a role in the genesis of a small subset of SCLC.      

NK/T细胞浸润对肺鳞癌患者生存与预后的影响

背景与目的:机体组织中自然杀伤细胞和细胞毒T淋巴细胞是构成机体细胞免疫的主导成分,对肿瘤组织必然有影响.本研究探讨NK/T细胞在肺鳞癌组织中的浸润程度及对患者生存与预后的影响.方法:将CD8作为细胞毒T淋巴细胞(CTL)的标记物,CD56作为自然杀伤细胞(NK)的特异性标记物,应用免疫组织化学方法检测肿瘤组织中NK/T细胞的分布和浸润情况.结果:68例肺癌中,CTL无/轻度浸润的者39例,5年生存率为18%;显著浸润者29例,5年生存率为42%;NK细胞无/轻度浸润者46例,5年生存率为14%,显著浸润者22例,5年生存率为45%;NK/T无轻度浸润者48例,5年生存率为33%,均显著浸润的20例,5年生存率为54%.经Log-rank检验,NK/T显著浸润组5年生存率显著高于无/轻度浸润组,差异有显著性(X2=18.62,P=0.00).结论:肺鳞癌组织中NK/T细胞显著浸润组的预后和生存时间明显优于轻度浸润组.其机制与机体细胞免疫有关.     

HLA-G expression in human ovarian carcinoma counteracts NK cell function.

BACKGROUND: Human leukocyte antigen-G (HLA-G) is an important immunotolerant which could be a part of the strategies applied by malignant cells applied to avoid host immunosurveillance. Aberrant expression of HLA-G has been found in ovarian carcinoma. The aim of this study was to evaluate the HLA-G expression in ovarian cancer tissues and to explore its function in vitro. MATERIALS AND METHODS: HLA-G expression in 33 primary ovarian carcinoma tissues was analyzed using immunohistochemistry with the anti-HLA-G monoclonal antibody (mAb) 4H84. Furthermore, the function of HLA-G in NK cell cytotoxicity was determined in vitro by cloning and expression of HLA-G on the ovarian carcinoma cell OVCAR-3. RESULTS: HLA-G expression was detected in 22/33 (66.7%) primary tumor tissues, but was absent in normal ovarian tissues (P<0.01). Cytotoxicity studies showed that HLA-G expression dramatically inhibits cell lyses by NK-92 cells (P<0.01), which could be restored by the anti-HLA-G conformational mAb 87G (P<0.01). CONCLUSION: HLA-G was expressed in a significant number of primary ovarian carcinoma tissues, and HLA-G expression in OVCAR-3 could directly inhibit NK-92 cell lysis. Taken together, our results indicated that expression of HLA-G plays an important role in evasion of ovarian cancer cells from host immunosurveillance.     

Distinct patterns of genetic alterations in adenocarcinoma and squamous cell carcinoma of the lung

Squamous cell carcinoma (SqC) and adenocarcinoma (AdC) are the two most common subtypes of non-small cell lung cancer (NSCLC). Cumulative information suggests that the SqC and AdC subtypes progress through different carcinogenic pathways, but the genetic aberrations promoting such differences remain unclear. Here we have assessed the overall genomic imbalances and structural abnormalities in SqC and AdC. By parallel analyses with comparative genomic hybridisation (CGH) on tumorous lung tissues and spectral karyotyping (SKY) on short-term cultured primary tumours, genome-wide characterisation was carried out on 69 NSCLC (35 SqC, 34 AdC). Molecular cytogenetic characterisation indicated common and distinct genetic changes in SqC and AdC. Common events of +1q21-q24, +5p15-p14, and +8q22-q24.1, and -17p13-p12 were found in both groups, although hierarchical clustering simulation on CGH findings depicted +2p13-p11.2, +3q25-q29, +9q13-q34, +12p, +12q12-q15 and +17q21, and -8p in preferential association with SqC pathogenesis (P<0.05). Corresponding SKY analysis suggested that these changes occur in simple and complex rearrangements, and further indicated the clonal presence of translocation partners leading to chromosomal over-representations. These recurring rearrangements involved chromosome pairs of t(1;13), t(1;15), t(7;8), t(8;15), t(8;9), t(2;17) and t(15;20). Of particular interest was the finding that the t(8;12) translocation partner was exclusive to AdC. The combined application of SKY and CGH has thus uncovered the genome-wide chromosomal aberrations in NSCLC. Specific chromosomal imbalances and translocation partners found in SqC and AdC have highlighted regions for further molecular investigation into gene(s) that may hold importance in the carcinogenesis of NSCLC.     

Receptor tyrosine kinase alterations and therapeutic opportunities in squamous cell carcinoma of the lung

Purpose

Targeted therapy has greatly improved the treatment for adenocarcinoma of the lung, but not squamous cell carcinoma (SCC) of the lung. The current paper describes the abnormalities of receptor tyrosine kinases (RTK) in lung SCC in a hope to stimulate the development of therapeutics that can have clinical impact.

Methods

We reviewed both clinical and preclinical studies published in English regarding RTK abnormalities and/or RTK-targeting treatment for SCC of the lung.

Results

RTK alterations have been demonstrated as biological signature for SCC of the lung. A number of clinical trials of RTK-targeting therapy have been carried out or are ongoing, with encouraging results.

Conclusions

SCC of the lung should be treated as an independent disease with unique treatment options based on molecular changes, particularly RTK.     

Endocrine function in small cell undifferentiated carcinoma of the lung

The endocrine status of 106 patients with undifferentiated small cell carcinoma of the lung was evaluated before treatment was begun. Almost one half of the patients had evidence of abnormal control of the secretion of adrenal cortical steroids, manifested by loss of diurnal rhythmicity or dexamethasone suppressibility. Only two had the clinical syndrome of ectopic ACTH secretion. Evidence of inappropriate secretion of vasopressin was found in 38% of the patients, most of whom also had abnormalities of corticosteroid secretory pattern. About one half of the patients had evidence of abnormal glucose tolerance, and many also had a paradoxical rise of plasma growth hormone concentration after glucose administration. The levels of the other hormones studies were normal. The pattern of hormone abnormality observed in these patients appears to be relatively specific for small cell undifferentiated carcinoma, and is different from that observed in other pulmonary tumors. Patients with abnormal control of plasma cortisol had a worse prognosis than those with normal adrenal function, largely because of decreased response rates to chemotherapy. Other endocrine abnormalities were of no prognostic significance.     

Clinical implications of intratumoral dendritic cell infiltration in esophageal squamous cell carcinoma

The clinical impact of intratumoral dendritic cell infiltration (IDCI) in esophageal cancer is not fully understood. We investigated the relationship between IDCI and clinical features in esophageal carcinoma. A retrospective analysis of a total of 203 patients who underwent esophagectomy for squamous cell esophageal carcinoma was performed. We evaluated IDCI immunohistochemically using S-100-protein. Under a high power objective (x400), we evaluated IDCI in 10 regions of surgical specimen including the most invasive slice and averaged the number of S-100-protein positive cells. The patients were classified into two groups according to IDCI (>20 or <20 S-100-protein positive cells per high power field: high or low IDCI, respectively). The degree of IDCI varied from 0 to 67 (average 12.3). The 203 patients were separated into 65 with high IDCI and 138 with low IDCI. The depth of invasion was significantly more superficial in patients with high IDCI (p<0.05), and the disease was at an earlier clinical stage (p<0.01) than in those with low IDCI. Five-year survival rates after curative resection were 67% and 39% in patients with high and low IDCI, respectively (p<0.01). Multivariate analysis showed that IDCI was an independent prognostic factor (RR=1.6, p=0.02), next to nodal involvement, depth of invasion and clinical stage. Being closely related to clinical features, the prognosis of patients with esophageal cancer may be estimated by monitoring IDCI.     

Prognostic significance of CpG island methylator phenotype in surgically resected small cell lung carcinoma

Methylation is closely involved in the development of various carcinomas. However, few datasets are available for small cell lung cancer (SCLC) due to the scarcity of fresh tumor samples. The aim of the present study is to clarify relationships between clinicopathological features and results of the comprehensive genome‐wide methylation profile of SCLC. We investigated the genome‐wide DNA methylation status of 28 tumor and 13 normal lung tissues, and gene expression profiling of 25 SCLC tissues. Following unsupervised hierarchical clustering and non‐negative matrix factorization, gene ontology analysis was performed. Clustering of SCLC led to the important identification of a CpG island methylator phenotype (CIMP) of the tumor, with a significantly poorer prognosis (P = 0.002). Multivariate analyses revealed that postoperative chemotherapy and non‐CIMP were significantly good prognostic factors. Ontology analyses suggested that the extrinsic apoptosis pathway was suppressed, including TNFRSF1A, TNFRSF10A and TRADD in CIMP tumors. Here we revealed that CIMP was an important prognostic factor for resected SCLC. Delineation of this phenotype may also be useful for the development of novel apoptosis‐related chemotherapeutic agents for treatment of the aggressive tumor.     

肝癌者红细胞和NK细胞的免疫活性研究

目的对肝癌患者的红、白细胞免疫粘附功能与T细胞亚群变化和NK细胞活性的关系进行观察.方法对53例肝癌患者在入院与病情恶化时,采用患者外周静脉血进行红、白细胞免疫粘附功能、T细胞亚群和NK细胞活性的对比测定.结果肝癌患者的红细胞C3b受体花环率、白细胞-补体受体花环率、CD3、CD4、CD8和CD4/CD8、及NK细胞的活性、第一次和第二次测定均低于对照组,P值分别为(P＜0.05)和(P＜0.01).同时还发现NK细胞的活性与红、白细胞的免疫功能密切相关.结论肝癌患者的红、白细胞免疫功能、T细胞亚群和NK细胞的活性低下,并随着病情恶化其降低显著扩大;NK细胞活性与红、白细胞的免疫功能密切相关.     

Microsatellite alterations in non-small cell lung cancer

Genetic alterations at 12 dinucleotide repeat loci located on human chromosomes 2, 3, 12, and 17 have been analyzed in non-small cell lung cancer from Thai patients. Seventeen out of 30 cases (57%) harbored the microsatellite alterations. Of the 30 cases, 19 patients had a history of tobacco smoking, of whom 14 (74%) were in the group with microsatellite alterations, whereas 3 out of 11 non-smokers (26%) had these alterations. The frequency of microsatellite alterations among smokers was significantly higher than it was in non-smokers (P = 0.01 Fisher's exact test; odds ratio; 7.47).     

Interleukin-8 in non-small cell lung carcinoma: relation with angiogenic pattern and p53 alterations

Progression of solid tumors, including NSCLC, is associated with increase in MVC (microvessel count), as a measure of tumor angiogenesis resulting from an imbalance between angiogenic factors and inhibitors. However, since tumor angiogenesis is a multi-step process under the control of various molecules, the mechanism of angiogenesis has not been fully clarified. Interleukin (IL)-8 has been shown to have a potential angiogenic effect in vitro and in vivo, and is overexpressed in several human solid cancers. Among the various angiogenic factors, vascular endothelial growth factor (VEGF) has been shown to correlate with a high MVC and with adverse prognosis in several human cancers, including NSCLC. Alterations of p53 suppressor gene are the most common genetic changes found in malignant tumors; several studies examined the link between aberrant p53 and angiogenesis in lung cancer, but only a few studies report data regarding a relation between p53 mutations and IL-8 expression. In this study we observed a correlation between IL-8 mRNA expression, intratumoral MVC and VEGF mRNA expression levels; furthermore, an aberrant p53 status was related to IL-8 expression. However, in our samples IL-8 levels did not significantly affect prognosis of NSCLC; more studies are required to elucidate the precise role of IL-8 in a large series of patients with non-small cell lung carcinoma.     

The intratumoral expression of vascular endothelial growth factor and interleukin-8 associated with angiogenesis in nonsmall cell lung carcinoma patients.

BACKGROUND: Angiogenesis has important effects on tumor growth and metastasis. It is regulated by a variety of angiogenic and angiostatic factors. METHODS: To evaluate the effects of tumor cell-derived angiogenic factors, we performed an immunohistochemic study to evaluate the intratumoral expression of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) in relation to intratumoral microvessel density (IMD) in tumors from 104 nonsmall cell lung carcinoma (NSCLC)patients. RESULTS: Fifty-four carcinomas were VEGF-positive, 47 carcinomas were IL-8-positive, and 53 carcinomas were hypervascular tumors. There was no significant correlation between the percentages of positive VEGF-staining and positive IL-8-staining in NSCLCs (rho = 0.174, P = 0.080). The IMD of VEGF-positive carcinomas was significantly greater than that of VEGF-negative carcinomas (P = 0.023). In addition, the IMD of IL-8-positive carcinomas was significantly greater than that of IL-8-negative carcinomas (P =0.013). The overall survival rate of patients with hypervascular tumors was significantly lower than that of patients with hypovascular tumors (41.0% versus 67.0%, P = 0.004). Cox proportional-hazards regression model also demonstrated that angiogenesis was one of the significant factors in predicting the survival of NSCLC patients (relative risk = 1.944, P = 0.041). CONCLUSIONS: Intratumoral expression of VEGF and IL-8 was associated with angiogenesis in NSCLCs. Tumor angiogenesis significantly affected the prognosis of NSCLC patients.     

Human breast cancer and impaired NK cell function

Recent advances in tumor immunology have led to the discovery of a new lymphoid cell with unique antitumor activity. Natural killer (NK) cells form an antitumor surveillance system and appear to be vital in preventing tumor growth and metastasis in animal models. We studied NK activity in patients with benign and malignant breast disease, using a chromium-51 release microtiter cytotoxicity assay with K562 cells as targets. Compared with benign controls, patients with malignancies had significantly depressed NK-mediated lysis (P less than 0.01). Furthermore, lysis in those with advanced disease (stages II, III, and IV) was significantly less than in those with limited disease (stage I) (P less than 0.01). NK activity was not correlated to estrogen or progesterone receptor states. Positive correlation of a depressed natural killer activity with the extent of tumor spread supports the concept of an NK cell immune surveillance system in breast cancer and emphasizes its importance in this malignancy.     

肺鳞状细胞癌分子特征及治疗对策研究现状

目的:总结肺鳞状细胞癌分子特征改变的研究现状及相关临床试验结果,探讨肺鳞癌的个体化治疗策略。方法:应用PubMed、CBM、CNKI和Medline数据库检索系统,以肺鳞状细胞癌、突变、过表达、扩增和临床试验及相关因子名称为关键词,检索2000-01-2012-08的相关文献。纳入标准:1)肺鳞癌细胞存在的分子生物学改变;2)相关靶向治疗药物的临床试验进展。符合分析的文献27篇。结果:肺鳞癌细胞的一组分子特征改变在肺鳞癌发生发展过程中发挥了重要作用,根据治疗靶点的不同,可分为膜受体、信号通路分子和转录因子3类,其中膜受体包括FGFR1、IGF-1R、EGFRvⅢ、PDGFRA、DDR2和MET;信号通路分子包括PIK3CA、AKT1、PTEN和B-RAF;转录因子包括SOX2和Nrf2;基于这些靶点的靶向治疗药物陆续进入临床试验阶段并取得了一定的进展。结论:在肺鳞癌的诊治中,分子特征的检测应受到更多的重视,分子特征改变的研究不仅有助于明确肿瘤的生物学特性,而且可以为患者选择更加合理的、个体化的治疗方案。     

Association of vascular endothelial growth factor expression with intratumoral microvessel density and tumour cell proliferation in human epidermoid lung carcinoma.

Vascular endothelial growth factor (VEGF) expression, vascularisation and tumour cell proliferation were analysed in 91 human epidermoid lung carcinomas using immunohistochemistry. A polyclonal anti-VEGF antibody was used for VEGF expression, a polyclonal antibody directed against human von Willebrand factor (factor VIII) to identify blood vessels and the proliferating cell nuclear antigen (PCNA) as a marker for proliferating cells. Positive staining for VEGF was obtained in 54 out of 91 cases (59%), the number of blood vessels varied from zero to 64 counts (mean 9.4) and the proportion of PCNA-positive cells varied from 1.3% to 72.1% (mean 25.2%). The mean PCNA labelling index and mean microvessel count in VEGF-positive tumours were significantly higher than those in VEGF-negative tumours (Wilcoxon rank sum test, P<0.0001; p<0.05). In addition, PCNA labelling index significantly increased with increasing VEGF expression (Jonckheere test, P<0.0001). In contrast, no association was found between PCNA labelling index and tumour vascularity (r=0.07, P=0.48). The close correlation of VEGF expression with tumour cell proliferation and microvessel density suggests that VEGF acts both as an autocrine growth factor and as stimulator for angiogenesis. However, tumour cell proliferation and microvessel growth and/or density may be regulated by separate mechanisms.     

Differential expression and function of the caveolin-1 gene in non-small cell lung carcinoma

This study was designed to clarify the function of caveolin-1 (Cav-1) in the development of lung cancer by investigating the mutation and protein expression of the Cav-1 gene in non-small cell lung carcinoma (NSCLC). Quantum dot immunofluorescence histochemistry was used to evaluate Cav-1 protein expression and subcellular localization in the lung cancer tissue microarray including 140 cases of lung cancer and 20 cases of non-cancerous lung tissue. Mutation of the Cav-1 gene in exon 1 and exon 3 was detected by polymerase chain reaction-single strand conformation polymorphism and sequencing. The positive rates of Cav-1 expression were 49.3% (69/140) in NSCLC group, significantly lower than the 100% (20/20) rate in the control group. Adenocarcinomas (16.7%), adenosquamous carcinomas (38.4%), squamous cell carcinomas (67.1%) and large cell lung cancers (66.7%) displayed Cav-1 positive staining, suggesting a gradient of Cav-1 expression according to tumor histotype-related aggressiveness. High-expression of Cav-1 protein was statistically correlated with pathologic TNM stage and lymph node metastasis. No mutation could be detected in exon 1 and exon 3 from all Cav-1 protein negative expression of NSCLC samples. Cav-1 immunoreactivity in lung cancer is histotype-dependent, increased Cav-1 expression indicates the malignant progression and high invasion features of NSCLCs. Deregulation of Cav-1 expression in NSCLCs may not correlate with mutation.