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  • Title: [Establishment of a novel chinese human lung adenocarcinoma cell line CPA-Yang3 and its real bone metastasis clone CPA-Yang3BM in immunodeficient mice].
    Author: Yang S, Shi M, Cao J, Su J, Zhao L, Lei B, Chang C, Lu J, Ye J, Xie W.
    Journal: Zhongguo Fei Ai Za Zhi; 2011 Feb; 14(2):79-85. PubMed ID: 21342638.
    Abstract:
    BACKGROUND AND OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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. 背景与目的: 肺癌复发和转移仍是世界性医学难题。本研究旨在建立国人肺腺癌转移细胞株，为研究肺癌转移的分子机制提供新颖的实验材料。 方法: 本细胞取自首诊双侧淋巴结转移，65岁女性肺腺癌(Ⅲb期)患者的胸水细胞原代培养即获成功。致瘤率实验，细胞生长曲线，染色体核型分析，荧光定量PCR检测，小鼠心室造模，核素骨显像和X片活体成像检测小鼠骨转移，取骨转移灶细胞原代培养，in vivo-in vitro重复多个cycles筛选骨转移细胞。 结果: 传自第4代细胞小鼠致瘤率达100%，经裸鼠左心室种植(1×106细胞/只)后第3周经核素骨显像、X片和病理检测确定骨转移率为：下颌骨100%，肱骨50%，股骨66.7%，脊柱50%，肩胛骨33%。同时伴有多脏器转移，肺(33%)、肝(50%)、肾上腺(17%)、颌下腺(33%)。荧光定量PCR检测ESM1、VEGF-C、IL-6、IL-8、AR、SVIL、FN1等基因过表达。将股骨转移细胞经体内外3个cycles筛选获得骨转移细胞CPA-Yang3BM。 结论: 新建的CPAYang3是个取自首诊双侧淋巴结转移肺腺癌病人的胸水原代细胞培养成功并经裸鼠心室造模、体内外4个cycles筛选后获得的多发骨转移的国人肺腺癌细胞株。
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