Yang Shunfang, Shi Meiping, Cao Jie, Su Jianzhong, Zhao Lanxiang, Lei Bei, Chang Cheng, Lu Jianying, Ye Jianding, Xie Wenhui
Department of Nuclear Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, China.
Zhongguo Fei Ai Za Zhi. 2011 Feb;14(2):79-85. doi: 10.3779/j.issn.1009-3419.2011.02.01.
The recurrence and metastasis of lung cancer is a tough problem worldwide. The aim of this study is to establish a novel Chinese lung adenocarcinoma cell line and its real bone-seeking clone sub-line for exploring the molecular mechanism of lung cancer metastasis.
The cells came from the pleural effusion of a sixty-five years old female patient with lung adenocarcinoma and supraclavicular lymph node metastases. The gene expression was detected by real-time quantitative PCR. Intracardiac injection of the cells into nude mice was performed and in vivo imaging was obtained by bone scintigraphy and conventional radiography. Bone metastases were determined on bone scintigraphy and then the lesions were resected under deep anesthesia for bone metastasis cancer cell culture. The process was repeated for four cycles to obtain a real bone-seeking clone.
The tumorigenesis rate started at 4th passage in immunodeficient mice via subcutaneously and as well as later passages. Approximately 1×10⁶ cancer cells were injected into left cardiac ventricle of immunodeficient mice resulted bone metastasis sites were successfully revealed by bone scintigraphy and pathological diagnosis, the mandible (100%), scapula (33%), humerus (50%), vertebral column (50%), femur (66.7%) and accompanied invasion with other organs, the adrenal gland (17%), pulmonary (33%), liver (50%), submaxillary gland (33%) in the mice after inoculation two-three weeks. The chromosome karyotype analysis of the cells was subdiploid. Quantitative real-time PCR was used to examined and compared with SPC-A-1 lung adenocarcinoma, ESM1, VEGF-C, IL-6, IL-8, AR, SVIL, FN1 genes were overexpress. The novel cell was named CPA-Yang3. The femur metastasis cell was repeated in vivo-in vitro-in vivo with three cycles and harvested a real bone metastasis clone. It was named CPA-Yang3BM.
Tne characteristics of novel strain CPAYang3 is a highly metastasis cell line of Chinese lung adenocarcinoma and CPA-Yang3BM is a real bone-seeking clone.
肺癌的复发和转移是全球范围内的一个难题。本研究旨在建立一种新型的中国肺腺癌细胞系及其真正的趋骨克隆亚系,以探索肺癌转移的分子机制。
细胞来源于一名65岁女性肺腺癌伴锁骨上淋巴结转移患者的胸腔积液。通过实时定量PCR检测基因表达。将细胞经心内注射到裸鼠体内,并通过骨闪烁显像和传统放射照相进行体内成像。通过骨闪烁显像确定骨转移,然后在深度麻醉下切除病变进行骨转移癌细胞培养。该过程重复四个周期以获得真正的趋骨克隆。
免疫缺陷小鼠在第4代皮下接种时开始致瘤,后续传代也有致瘤性。将约1×10⁶个癌细胞注入免疫缺陷小鼠左心室后,骨闪烁显像和病理诊断成功显示骨转移部位,下颌骨(100%)、肩胛骨(33%)、肱骨(50%)、脊柱(50%)、股骨(66.7%),并伴有其他器官浸润,接种后两至三周小鼠体内肾上腺(17%)、肺(33%)、肝(50%)、颌下腺(33%)。细胞染色体核型分析为亚二倍体。采用定量实时PCR检测并与SPC-A-1肺腺癌比较,ESM1、VEGF-C、IL-6、IL-8、AR、SVIL、FN1基因过表达。该新细胞命名为CPA-Yang3。股骨转移细胞经体内-体外-体内三个周期重复培养,获得真正的骨转移克隆。命名为CPA-Yang3BM。
新型细胞系CPAYang3的特点是中国肺腺癌高转移细胞系,CPA-Yang3BM是真正的趋骨克隆。