Establishment and characterization of four malignant pleural mesothelioma cell lines from Japanese patients
Establishment and characterization of four malignant pleural mesothelioma cell lines from Japanese patients
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DOI：
10.1111/j.1349-7006.2006.00184.x
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发表时间：
2006-05-01
期刊：
影响因子：
5.7
通讯作者：
Hida, T
中科院分区：
文献类型：
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作者：
Usami, N;Fukui, T;Hida, T
恶性胸膜间皮瘤（MPM）是一种石棉相关的恶性肿瘤，对目前的治疗方式具有高度抵抗力。我们从日本患者中建立了四种MPM细胞系（ACC-MESO-1、ACC-MESO-4、Y-MESO-8A和Y-MESO-8D），后两种来自同一患者，具有MPM的双相样特征，在细胞培养中分别显示上皮和肉瘤表型。这些细胞在添加10%胎牛血清的RPMI-1640培养基中在5%CO2下生长良好。突变和表达分析表明，肿瘤抑制基因NF 2，这是已知的MPM中最常见的突变之一，在ACC-MESO-1突变。我们在所有四个MPM细胞系中检测到p16（INK 4A）/p14（ARF）的纯合性缺失。然而，在这些细胞系中未发现其他肿瘤抑制基因（包括TP 53）和原癌基因（包括KRAS、NRAS、BRAF、EGFR和HER 2）的突变。聚合酶链反应扩增猴病毒40序列未检测到任何产物。我们还分析了其他6个MPM细胞系的遗传改变，并证实了NF 2和p16（INK 4A）/p14（ARF）的频繁突变。为了表征Y-MESO-8A和Y-MESO-8D之间的生物学差异，我们进行了cDNA微阵列分析，并检测在这两个细胞系中差异表达的基因。因此，我们的新MPM细胞系似乎是有用的新模型，用于研究人类MPM生物学的各个方面，以及材料的发展，未来的治疗。
Malignant pleural mesothelioma (MPM) is an asbestos-related malignancy that is highly resistant to current therapeutic modalities. We established four MPM cell lines (ACC-MESO-1, ACC-MESO-4, Y-MESO-8A and Y-MESO-8D) from Japanese patients, with the latter two from the same patient with biphasic-like characteristics of MPM, showing epithelial and sarcomatous phenotypes, respectively, in cell culture. These cells grew well in RPMI-1640 medium supplemented with 10% fetal bovine serum under 5% CO2. Mutation and expression analyses demonstrated that the tumor suppressor gene NF2, which is known to be one of the most frequently mutated in MPM, is mutated in ACC-MESO-1. We detected homozygous deletion of p16(INK4A)/p14(ARF) in all four MPM cell lines. However, mutations of other tumor suppressor genes, including TP53, and protooncogenes, including KRAS, NRAS, BRAF, EGFR and HER2, were not found in these cell lines. Polymerase chain reaction amplification of the simian virus 40 sequence did not detect any products. We also analyzed genetic alterations of six other MPM cell lines and confirmed frequent mutations of NF2 and p16(INK4A)/p14(ARF). To characterize the biological differences between Y-MESO-8A and Y-MESO-8D, we carried out cDNA microarray analysis and detected genes that were differentially expressed in these two cell lines. Thus, our new MPM cell lines seem to be useful as new models for studying various aspects of the biology of human MPM as well as materials for the development of future therapies.