中国人肺腺癌细胞系CPA-Yang2的建立及其生物学特性

Establishment and Characterization of a Novel Chinese Human Lung Adenocarcinoma Cell Line CPA-Yang2 in Immunodeficient Mice

背景与目的肺癌的复发、转移和耐药是肺癌治疗的难题。本研究旨在建立国人肺腺癌转移细胞株,为肺癌的发生、发展和耐药的研究提供新的实验模型。方法本细胞取自化疗10次、放疗1次临床认定均无效的56岁女性肺腺癌患者的腹水且放射免疫法肿瘤标志物检测CA125、CYFRA21-1、CEA和NSE均超标,细胞原代培养即获成功。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,荧光定量PCR检测部分基因表达。结果传自第8代细胞经裸小鼠皮下种植致瘤率仅30%,传自第11代且以后各代均表现稳定的致瘤率(100%),体外培养细胞在显微镜下显示为贴壁生长,该细胞的染色体核型分析为亚三倍体;经裸鼠左心室或尾静脉分别接种1×106个细胞(0.1mL)、1.5×106个细胞(0.15mL),2周后陆续出现小鼠体重下降,第3周经核素骨显像检测和病理确定为骨转移达100%,同时伴有各脏器转移:脑(30%)、肺(50%)、肝(60%)和肾上腺(80%)。荧光定量PCR检测ESM1、VEGF-C、IL-6、IL-8、AR等基因过表达。结论新建的CPA-Yang2是取自化疗10次、放疗1次均NR的肺腺癌病人腹水培养成功并经裸鼠血道种植后具有除多发骨转移外同时伴有脑、肝、肾上腺和肺转移特性的国人肺腺癌细胞株,细胞代谢旺盛,性状稳定。该细胞将成为国人肺癌研究的又一个良好细胞实验模型。

Background and objective The recurrence, metastasis and multidrug resistance （MDR） in lung cancer are the tough problems worldwide. This study was to establish a novel chinese lung adenocarcinoma cell line with high metastasis potential for exploring the mechanism ofreccurrence, development and MDRin lung cancer. Methods The cell came from the abdominal dropsy of a fifty-six years old female patient with lung adenocarcinoma and the tumor markers CA125, CYFRA21-1, CEA, NSE were detected to be higher secretion by radioimmunoassay in the abdominal dropsy. Tumorigenicity of immunodeficient mice was confirmed in 8th passage. The cell growth curve was mapped. Analysis of chromosome karyotype was tested. The gene expression was measured by real-time quantitative PCR. Results The tumorigenesis rate started at 8th passage in 3/10 immunodeficient mice via subcutaneously and the fully tumorigenicity was at 1 lth passage as well as later passages. Under the microscope, the cell showed oval-shap and adherence. The chromosome karyotype analysis of the ceils was sub-triploid. Approximately 1 × 10^6 and 1.5× 10^6 cancerous cells were injected into left cardiac ventricle and tail vein of immunodeficient mice respectively, The results showed multiorgan metastasis in the mice after three-four weeks, including mandible, scapula, humerus, vertebral column, femur, rib and brain, liver, adrenal gland, pulmonary in the mice after inoculation. The bone metastasis rate was 100% in the tumor bearing mice by bone scintigraphy and pathology. Quantitative real-time PCR was used to examined and compared with SPC-A-1 lung adenocarcinoma, ESM1, VEGF-C, IL-6, IL-8, AR genes were overexpressed. The novel cell was named CPA-Yang2. Conclusion The characteristics of novel strain CPA-Yang2 is a highly metastasis cell line of Chinese lung adenocarcinoma. It has stable traits, highly metastasis ability and maybe is a MDR lung cancerous cell line. Of course, it＇s a good experimental model for lung cancer research.