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人胃癌细胞系的建立与鉴定

Establishment and characterization of human gastric carcinoma cell lines.

作者信息

Park J G, Yang H K, Kim W H, Chung J K, Kang M S, Lee J H, Oh J H, Park H S, Yeo K S, Kang S H, Song S Y, Kang Y K, Bang Y J, Kim Y H, Kim J P

机构信息

Laboratory of Cell Biology, Cancer Research Institute, Korean Cell Line Bank, Korean Cell Line Research Foundation, Seoul, Republic of Korea.

出版信息

Int J Cancer. 1997 Feb 7;70(4):443-9. doi: 10.1002/(sici)1097-0215(19970207)70:4<443::aid-ijc12>3.0.co;2-g.

Abstract

We report 8 newly established gastric-carcinoma cell lines (SNU-216, 484, 520, 601, 620, 638, 668, 719) from Korean patients. Morphologic study was carried out using light and electron microscopes. CEA, alpha FP, and CA 19-9 and TPA in supernatant and in cell lysate were measured by radioimmunoassay. p53 and c-Ki-ras gene mutations were screened and confirmed by sequencing. The cell lines, derived from tumors with moderate differentiation, grew as a diffuse monolayer, and those from tumors with poor differentiation and minimal desmoplasia grew exclusively as non-adherent. Out of the 8 gastric-cancer cell lines, 5 had detectable levels of CEA both in supernatant and in cell lysate; there was no expression or secretion of alpha FP in these cells; 4 cell lines showed high levels of CA 19-9 in cell pellets. All cell lines except SNU-484 had high concentrations of TPA both in cell lysate and in supernatants. p53 mutation was found in 6 cell lines (75%): 2 (SNU-216 and SNU-668) had mutations in exon 6, and other 3 in exon 8. The c-Ki-ras mutation was found in 2 cell lines (25%), SNU-601 and SNU-668. The former showed GGT-to-GAT transition mutation at codon 12, while the latter showed CAA-to-AAA transversion mutation at codon 61. DNA profiles using restriction endonuclease HinfI and polymorphic DNA probes ChdTC-15 and ChdTC-114 showed different unique patterns; which suggests that these cell lines are unique and not cross-contaminated. We believe that the newly characterized gastric-cancer cell lines presented in this paper will provide a useful in vitro model for studies related to human gastric cancer.

摘要

我们报告了从韩国患者中新建的8种胃癌细胞系(SNU - 216、484、520、601、620、638、668、719)。利用光学显微镜和电子显微镜进行了形态学研究。通过放射免疫分析法检测了上清液和细胞裂解物中的癌胚抗原(CEA)、甲胎蛋白(αFP)、糖类抗原19 - 9(CA 19 - 9)和组织多肽抗原(TPA)。通过测序筛选并确认了p53和c - Ki - ras基因突变。这些细胞系来源于中度分化的肿瘤,呈弥漫性单层生长,而来源于低分化且极少伴有纤维组织增生的肿瘤细胞系则仅以非贴壁方式生长。在这8种胃癌细胞系中,5种在上清液和细胞裂解物中均可检测到CEA水平;这些细胞中未检测到αFP的表达或分泌;4种细胞系在细胞沉淀中显示出高水平的CA 19 - 9。除SNU - 484外,所有细胞系在细胞裂解物和上清液中均含有高浓度的TPA。在6种细胞系(75%)中发现了p53突变:2种(SNU - 216和SNU - 668)在外显子6发生突变,另外3种在外显子8发生突变。在2种细胞系(25%)SNU - 601和SNU - 668中发现了c - Ki - ras突变。前者在密码子12处发生GGT到GAT的转换突变,而后者在密码子61处发生CAA到AAA的颠换突变。使用限制性内切酶HinfI以及多态性DNA探针ChdTC - 15和ChdTC - 114进行的DNA图谱分析显示出不同的独特模式;这表明这些细胞系是独特的,且未发生交叉污染。我们认为本文中所描述的新鉴定的胃癌细胞系将为人类胃癌相关研究提供一个有用的体外模型。

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