Differential Tks5 isoform expression contributes to metastatic invasion of lung adenocarcinoma

Metastasis accounts for the vast majority of cancer-related deaths, yet the molecular mechanisms that drive metastatic spread remain poorly understood. Here we report that Tks5, which has been linked to the formation of proteolytic cellular protrusions known as invadopodia, undergoes an isoform switch during metastatic progression in a genetically engineered mouse model of lung adenocarcinoma. Nonmetastatic primary tumor-derived cells predominantly expressed a short isoform, Tks5_short, while metastatic primary tumor- and metastasis-derived cells acquired increased expression of the full-length isoform Tks5_long. This elevation of Tks5_long to Tks5_short ratio correlated with a commensurate increase in invadopodia activity in metastatic cells compared with nonmetastatic cells. Further characterization of these isoforms by knockdown and overexpression experiments demonstrated that Tks5_long promoted invadopodia in vitro and increased metastasis in transplant models and an autochthonous model of lung adenocarcinoma. Conversely, Tks5_short decreased invadopodia stability and proteolysis, acting as a natural dominant-negative inhibitor to Tks5_long. Importantly, high Tks5_long and low Tks5_short expressions in human lung adenocarcinomas correlated with metastatic disease and predicted worse survival of early stage patients. These data indicate that tipping the Tks5 isoform balance to a high Tks5_long to Tks5_short ratio promotes invadopodia-mediated invasion and metastasis.     

BRI基因在一对转移能力不同的肺腺癌细胞系中的序列分析与表达研究

目的 确认淀粉样纤维蛋白基因(amyloid fibrils，BRI)基因在l对同源但转移能力不同的肺腺癌细胞系AGZY83-a和Anip973中的序列并分析其表达。方法 采用测序技术，Northern印迹杂交。G显带后荧光原位杂交分析BRI基因在肺腺癌细胞系的序列与表达。结果 BRI基因在高转移肺腺癌细胞系Anip973中高表达，在其低转移母系AGZY83-a中低表达，两细胞系BRI基因染色体定位区均存在断裂重排。该基因染色体定位区在Anip973中出现扩增。已知BRI基因的-116bp～-5bp处碱基序列和-115bp～-5bp处碱基序列在AGZY83-a和Anip973中分别突变为CTCAGCAGCCCGC和TCAGCCGC。结论 BRI基因在转移能力不同的肺腺癌细胞系差异表达与该基因的染色体定位区域的断裂重排无关，与该基因染色体定位区拷贝数增加及5′非翻译区存在不同的突变可能相关。     

MicroRNA-130a inhibits cell proliferation,invasion and migration in human breast cancer by targeting the RAB5A

MiR-130a has been demonstrated to play important roles in many types of cancers. Nevertheless, its biological function in breast cancer remains largely unknown. In this study, we found that the expression level of miR-130a was down-regulated in breast cancer tissues and cells. Overexpression of miR-130a was able to inhibit cell proliferation, invasion and migration in MCF7 and MDA-MB-435 cells. With the bioinformatics analysis, we further identified that RAB5A was a directly target of miR-130a, and its mRNA and protein level was negatively regulated by miR-130a. Immunohistochemistry verified RAB5A was upregulated in breast cancer tissues. Therefore, the data reported here demonstrate that miR-130a is an important tumor suppressor in breast cancer, and imply that miR-130a/RAB5A axis have potential as therapeutic targets for breast cancer.     

Increased expression of Rab5A predicts metastasis and poor prognosis in colorectal cancer patients

Rab5A is reported to correlate with cancer development and progression. The purpose of this study is to explore the association between Rab5A expression and the clinical characteristics of colorectal cancer (CRC). Data containing three independent investigations from Oncomine database demonstrated that Rab5A is overexpression in CRC compared with normal tissue, similar result was also found in 32 matched CRC tissue samples by qPCR. The protein expression of Rab5A was examined in 390 CRC specimens and the results showed that high expression of Rab5A was significantly correlated with tumor size (P = 0.008), serum CEA (P = 0.002), liver metastasis (P = 0.014) and clinical stage (P = 0.010). Kaplan-Meier method suggested that overexpression of Rab5A protein expression had shorter overall survival times in CRC patients (P < 0.001). Multivariate Cox regression analysis confirmed Rab5A expression, tumor size and clinical stage as independent prognostic factor in CRC. In conclusion, the data indicated that higher expression of Rab5A was observed in CRC tissues and Rab5A may be identified as a useful predictor of metastasis and prognosis for CRC.     

基因芯片检测人肺鳞癌和肺腺癌基因表达的异同

目的用基因芯片技术检测肺鳞癌和肺腺癌基因表达的异同。方法提取人肺鳞癌和肺腺癌组织及正常肺组织的RNA,分别用Cy5-dCTP或Cy3-dCTP标记,再与4096点基因芯片杂交,检测肺鳞癌和肺腺癌组织基因表达的异同。结果肺鳞癌和肺腺癌表达共同上调的基因17条,共同下调的基因19条;肺鳞癌表达显著高于肺腺癌的基因20条,显著低于肺腺癌的基因14条。结论多基因参与肺癌发病,基因芯片技术是肺癌基因表达检测的有效方法。     

5-氟尿嘧啶诱导人肺腺癌A549细胞自噬

目的 探讨5-氟尿嘧啶(5-FU)是否诱导人肺腺癌A549细胞发生自噬.方法 吖啶橙(AO)荧光染色和透射电镜观察自噬泡的变化;细胞免疫化学法测定LC3-Ⅱ的表达;流式细胞术及吖啶橙染色结合激光扫描共焦显微镜检测细胞凋亡;DNA电泳检测凋亡梯形条带的产生.结果 吖啶橙荧光染色和透射电镜观察结果显示,5-氟尿嘧啶处理细胞...     

Mechanisms of lymphatic metastasis in human colorectal adenocarcinoma

The invasion of lymphatic vessels by colorectal cancer (CRC) and its subsequent spread to draining lymph nodes is a key determinant of prognosis in this common and frequently fatal malignancy. Although tumoural lymphangiogenesis is assumed to contribute to this process, review of the current literature fails to support any notion of a simple correlation between lymphatic vessel density and CRC metastasis. Furthermore, attempts to correlate the expression of various lymphangiogenic growth factors, most notably VEGF‐C and VEGF‐D, with the lymphatic metastasis of CRC have provided contradictory results. Recent evidence from animal and human models of tumour metastasis suggests that complex functional and biochemical interactions between the microvasculature of tumours and other cell types within the tumour microenvironment may play a pivotal role in the behaviour of commonly metastasizing tumours. Indeed, previous insights into tumoural blood vessels have provided candidate markers of tumoural angiogenesis that are currently the subject of intense investigation as future therapeutic targets. In this review article we survey the current evidence relating lymphangiogenesis and lymphangiogenic growth factor production to metastasis by CRC, and attempt to provide some insight into the apparent discrepancies within the literature. In particular, we also discuss some new and provocative insights into the properties of tumoural lymphatics suggesting that they have specific expression profiles distinct from those of normal lymphatic vessels and that appear to promote metastasis. These findings raise the exciting prospect of future biomarkers of lymphatic metastasis and identify potential targets for new generation anti‐tumour therapies. Copyright © 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Pulmonary metastasis of cribriform‐morular variant of thyroid carcinoma mimicking primary adenocarcinoma of the lung: A potential pitfall

Cribriform‐morular variant of papillary thyroid carcinoma (CMV‐TC) shows a peculiar mixture of follicular, cribriform, papillary, trabecular, and solid patterns with squamoid morules. Ocassionally, lung metastasis may be interpreted incorrectly as primary lung adenocarcinoma. We illustrate a case of pulmonary meastasis of CMV‐TC mimicking a primary adenocarcinoma, 7 years after diagnosis of CMV‐TC. The lung metastases may be easily missed if the pathologist is unaware of the patient's prior history and a limited immunohistochemical panel (CK7 and TTF‐1) is used. The histologic and immunohistochemical (β‐catenin+, ER+, PR+, TTF‐1 +, and CK7+) findings were diagnostic of CMV‐TC and ensured adequate treatment.     

A RAB27A duplication in several cases of Griscelli syndrome type 2: An explanation for cases lacking a genetic diagnosis

Griscelli syndrome type 2 (GS2) is a rare and often fatal autosomal recessive, hyperinflammatory disorder. It is associated with hypopigmentation of the skin and the hair, resulting in the characteristic pigment accumulation and clumping in the hair shaft. Loss‐of‐function mutations in RAB27A, resulting from point mutations, short indel, or large deletions, account for all the cases reported to date. However, several GS2 cases originating from Saudi Arabia lack a genetic diagnosis. Here, we report on a new RAB27A genetic anomaly observed in seven Saudi Arabia families that had remained negative after extensive molecular genomic DNA testing. Linkage analysis and targeted sequencing of the RAB27A genomic region in several of these patients led to the identification of a common homozygous tandem duplication of 38 kb affecting exon 2–5 and resulting in a premature stop codon. The pathogenic effect of this duplication was confirmed by a cDNA analysis and functional assays. The identification of microhomology flanking the breakpoint site suggests a possible underlying mechanism.     

Differential gene expression in stromal cells of human giant cell tumor of bone

Giant cell tumor (GCT) offers a unique model for the hematopoietic–stromal cell interaction in human bone marrow. Evidence has been presented that GCT stromal cells (GCTSCs) promote accumulation, size and activity of the giant cells. Although GCTSCs are considered the neoplastic component of GCT, little is known about their genetic basis and, to date, a tumor-specific gene expression pattern has not been characterized. Mesenchymal stem cells (MSCs) have been identified as the origin of the GCT neoplastic stromal cell. Using state of the art array technology, expression profiling was applied to enriched stromal cell populations from five different GCTs and two primary MSCs as controls. Of the 29 differentially expressed genes found, 25 showed an increased expression. Differential mRNA expression was verified by real-time polymerase chain reaction analysis of 10 selected genes, supporting the validity of cDNA arrays as a tool to identify tumor-related genes in GCTSCs. Increased expression of two oncogenes, JUN and NME2, was substantiated at the protein level, utilizing immunohistochemical evaluation of GCT sections and Western-blot analysis. Increased phosphorylation of JUN Ser-63 was also found.     

不同转移潜能的人肿瘤细胞系金属蛋白酶活性分析

目的探讨不同转移潜能的人类肿瘤细胞的金属蛋白酶（ＭＭＰｓ）活性与其侵袭转移潜能的相关性。方法分别选取具有不同转移潜能的人类肺癌细胞系（ＰＧ、ＰＡａ、ＢＥ１、ＣＬ３、ＬＨ７）、黑色素瘤细胞系（ＷＭ３５、ＷＭ１３４１ｂ、ＷＭ９８３ａ、ＷＭ４５１）及前列腺癌细胞系（ＰＣ，ＰＣ３Ｍ），经细胞培养，条件培养基的收集与浓缩，利用明胶Ｚｙｍｏｇｒａｐｈｙ法检测以上各组细胞ＭＭＰ２和ＭＭＰ９的产生及活性差异，并将这种差异与各自的转移潜能联系起来。结果转移能力相对较高的细胞系产生ＭＭＰｓ的能力相应强于转移能力相对较低者：ＰＧ高于ＰＡａ，ＢＥＩ高于ＣＬ３和ＬＨ７。在进展期黑色素瘤株ＷＭ９８３ａ和转移瘤株ＷＭ４５１出现了ＭＭＰ９的表达，早期瘤株则无。前列腺瘤细胞ＰＣ３的转移性克隆ＰＣ３Ｍ的条件培养基中有较高的ＭＭＰ９活性。结论肿瘤细胞的侵袭转移潜能与其产生ＭＭＰｓ的能力密切相关。     

人非小细胞肺癌组织中Ku80 mRNA表达评价化学治疗敏感性的研究

目的：通过观察Ku80 mRNA在人正常肺组织、未经化疗和多次化疗的非小细胞肺癌组织的定量表达情况，探讨Ku80是否可以预测和评价肺癌化学治疗的敏感性。方法：采用RT-PCR方法，以B-actin为内参照，检测、比较25例正常肺组织和51例肺癌患者肺组织中Ku80 mRNA的半定量表达，以SPSS统计软件进行分析。结果：比较人正常肺组织组与未经化疗肺癌组和多次化疗肺癌组Ku80 mRNA表达量的差别具有统计学意义，分别为P〈0．05和P〈0．01。未经化疗肺癌组与多次化疗肺癌组（n≥2）的Ku80 mRNA的表达量相比具有统计学意义（P〈0．05）。结论：正常肺组织和肺癌组织在Ku80 mRNA表达量上具有统计学差异，并且Ku80 mRNA表达量在肺癌组织中随化疗次数增加明显上升，提示Ku80 mRNA定量表达可能成为对肺癌化疗的敏感性的预测和评价提供新的依据。     

肺腺癌细胞微环境及转移与黏附分子表达的关系

目的研究肺腺癌细胞生长环境及转移性与黏附分子CD44v6和CD29的表达关系。方法将起源相同、转移性不同的两个肺腺癌细胞系AGZY和Anip分别用简便肿瘤多细胞球体(MTS)培养法培养，并设常规单层贴壁细胞培养对照。通过倒置显微镜、扫描及透射电镜观察MTS形成情况，并用免疫组化法分别对MTS及贴壁细胞上CD44v6和CD29表达进行检测。结果MTS培养成功，贴壁细胞与MTS在细胞结构及细胞连接结构上相似，两种MTS在形态及结构上差异无显著性。免疫组化结果显示，CD29在高转移性的Anip细胞及其MTS上呈阳性表达；在低转移性的AGZY细胞及其MTS上阴性表达。CD44v6在Anip和AGZY细胞及MTS上均呈阳性表达，差异无显著性。贴壁细胞与MTS上两种黏附分子表达均无差异。结论成功建立了一种简易制备MTS的方法。细胞生长方式(单层贴壁与MTS)可能不影响CD44v6和CD29的表达。CD29表达可能与肺腺痛转移性相关；CD44v6表达可能与肺腺癌转移无关。     

Analysis of RAB27A Gene in Griscelli Syndrome type 2: Novel Mutations Including a Deletion Hotspot

INTRODUCTION: Griscelli syndrome type 2 is an autosomal recessive disorder characterized by pigmentary dilution and occurrence of acute phases of hemophagocytosis. The disease is caused by mutations in RAB27A gene, coding a small GTPase involved in terminal phases of cytotoxic granule/melanosome exocytosis. MATERIALS AND METHODS: We describe the result of mutation analysis among nine patients from seven non-related Persian families. We present four novel mutations including a deletion hot spot (514del 5). CONCLUSION: This hot spot is flanked by "direct repeats" of nucleotides, which are previously shown to be associated with areas of recurrent small deletions.     

ALK protein expression in pulmonary adenocarcinoma of Tunisian patients

Background. It is now necessary to determine ALK status in order to use targeted therapy. Aim: herein, we assess immunohistochemical profile of ALK protein in a series of Tunisian patients with pulmonary adenocarcinoma.

Materials and Methods. ALK protein expression was studied applying the D5F3 antibody with a fully automated Ventana CDx technique on a series of 19 patients.

Results. Positive ALK expression was found in one case (5.2%) corresponding to a papillary adenocarcinoma which showed a strong granular and homogenous cytoplasmic staining. The patient was a 30-years-old woman.

Conclusion. The frequency of positive ALK expression based on immunohistochemistry in our series was similar to that reported in the world literature.     

人肝癌组织nm23的表达及其与转移的关系

目的：研究ｎｍ２３基因在肝癌转移，发生，发展中的意义，方法：应用免疫组人技术（ＳＰ法）检测ｎｍ２３在２４例人肝癌组织的表达，结果ｎｍ２３在肝癌组织中较高表达，在癌旁组织中也有表达，在已发生临床转移的肝癌组织中ｎｍ２３的表达水平明显低于尚未发生转移，结论：ｎｍ２３在抑制肝癌转移中有一定的作用，但在肝癌转移过程显然尚有其他调节因素的存在。