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Molecular cloning of the genes related to the early stage of the peripheral type of lung adenocarcinomas

周围型肺腺癌早期相关基因的分子克隆

基本信息

批准号：
12670215
负责人：
YAMAZAKI Hitoshi
金额：
$ 2.3万
依托单位：
Tokai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

In this study, we tried to identify some functioning molecules which would be related to carcinogenesis and development of lung adenocarcinomas by way of differential screening of early lung cancer. Recently, the specimens of the early stage of peripheral lung adenocarcinomas have been available as endoscopic resection of lung cancer are performed. We collected 40 cases of small lung tumors and their unaffected normal lung tissues. Detail histological analysis revealed that these samples are categorized as bronchioloalveolar adenocarcinomas and invasive adenocarcioma. Total RNA was extracted, reverse-transcribed with Rhodamine-labeled oligo-d(T) and applied for PCR in combination with 24 kinds of forward primers. Total number of combination is 216. The samples were electrophoresed on a sequencing gel. The interested bands were identified by fluorescent bioimage analyzer (FMBIO-II Multi View, Takara, Kyoto) and cut out from the gel. The cDNA was isolated from the gel, purified and directly sequenced. After the first step of cloning, total 323 clones were applied for direct sequencing. Finally 255 clones(163 clones possess poly(A)^+ and 92 clones do not possess poly(A)^+) were taken for the further analysis. Now we are searching the interested candidates in these clones by using BLAST analysis method. In this study we also analyze 1L-10 gene expression in some non-small cell lung cancers (NSCLC). We concluded that IL-10 expression was related to the clinical prognosis of NSCLC patients and also related to the gene expression of angiopoietin system (Ang-1, Ang-2, Angiopoietin Receptor)

本研究试图通过对早期肺癌的鉴别筛查，寻找一些与肺腺癌发生发展相关的功能分子。近年来，随着对肺癌的内窥镜切除，早期周围型肺腺癌的标本已经有了。我们收集了40例肺小肿瘤及其正常肺组织。详细的组织学分析显示，这些标本被归类为细支气管腺癌和浸润性腺癌。提取总RNA，用罗丹明标记的寡核苷酸(T)反转录，与24种正向引物结合应用于聚合酶链式反应。组合总数为216个。这些样品在测序凝胶上进行了电泳法。用荧光生物图像分析仪(FMBIO-II多视点，Takara，京都)鉴定感兴趣的条带，并从凝胶中切下。从凝胶中分离、纯化并直接测序。经第一步克隆后，共323个克隆进行直接测序。最后对255个克隆进行进一步分析，其中163个克隆具有PolyA^+，92个克隆不具有PolyA^+。现在我们正在用BLAST分析方法在这些克隆中寻找感兴趣的候选基因。本研究还分析了一些非小细胞肺癌(NSCLC)中1L-10基因的表达。提示IL-10的表达与非小细胞肺癌患者的临床预后有关，也与血管生成素系统(Ang-1、Ang-2、Angiopoietin受体)的基因表达有关。