背景与目的 肺癌是我国肿瘤疾病中的头号杀手.建立国人肺腺癌高转移细胞株CPA-Yang1,为肺癌的发生、发展的研究提供实验模型.方法 本细胞取自50岁男性肺腺癌患者的心包积液作细胞原代培养成功.免疫缺陷小鼠致瘤率实验;绘制细胞生长曲线;细胞染色体核型分析;肿瘤标志物检测;荧光定量PCR检测部分癌基因表达.结果 第一代细胞经BNX小鼠和裸鼠皮下种植的动物实验证实,瘤体生长迅速,致瘤率达100%且以后各代均表现稳定的强致瘤率(100%);体外培养细胞在显微镜下显示为半悬浮生长且细胞面积较大;经裸鼠左心室和尾静脉种植(75-80万细胞/只)2-3周后陆续出现小鼠下肢瘫痪和脊柱骨肿胀变形且体重下降,经核素骨显像检测和病理确定为骨转移达90%,且仅10%同时伴有肺转移.该细胞的染色体核型分析为亚三倍体;肿瘤标志物放射免疫检测CEA为高分泌(10.1 ng/mL).RT-PCR检测VEGF-C、IL-6、IL-8癌基因过表达.结论 新建的CPA-Yang1是株亲代细胞就具有骨转移特性的国人肺腺癌细胞株,细胞代谢旺盛,性状稳定,细胞形态特殊,该细胞将成为肺癌研究的一个良好细胞实验模型.%Background and objective Lung cancer is a common malignancy and is the major determinant of overall cancer mortality worldwidely. Approximately 70％ of lung cancer patients will die from metastatic diseases. The aim of this study is to establish a Chinese lung adenocarcinoma cell line with high metastasis potency for exploring the mechanism of occurrence and development in lung cancer. Methods The cell came from the pericardial effusion of a fifty-year old male patient with lung adenocarcinoma and the cells in primary culture were obtained successfully. Immunodeficient mice tumorigenidty was assayed. The cell growth curve was mappinged Analysis of chromosome karyotype was tested. Tumor marker was detected by radioim-munoassay.The gene expression was measured by real-time quantitative PCR. Results The first passage cells were planted in im-munodeficiant mice subcutaneously and the tumorigenesis rate was 100％ as well as later passages. Under the microscope, the cell showed larger and semi-suspension, semi-adherence. Approximately 0.8x 106 cancerous ceils were injected into left cardiac ventri-cle or tail vein of immunodeficient mice resulted start to appear lower limb paralysis and spine swelling deformation in the mice after inoculation two-three weeks. The bone metastasis rate was 90％ in the tumor bearing mice by bone acintigraphy and pathol-ogy and only pulmonary metastasis 10％ at the same time. The chromosome karyotype analysis of the cells was sub-triploid. The tumor marker CEA was detected in higher secretion by radioimmunoassay in the cell culture suspension. Quantitative real-time PCK was used to examined and compared SPC-A-1 lung adenocarcinoma, VEGF-C, IL-6,IL,-8, genes were overexpressed. The novel cell line was named CPA-Yang1. Conclusion Tne novel strain CPA-Yang1 is an parental cell with characteristics of bone metastasis of Chinese lung adenocarcinoma. It has stable traits, highly metastatic ability and a good experimental modal for lungcancer research.
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