人大细胞肺癌细胞系NL9980和L9981的建立及其生物学特性研究

Establishment and their biological characteristics of clonal cell subpopulations (NL9980 and L9981) from a human lung large cell carcinoma cell line (WCQH-9801)

目的 探讨从人大细胞肺癌细胞株WCQH 980 1分离构建不同转移潜能肺癌细胞株的可能性 ,并鉴定其生物学和分子生物学差异。方法 应用单个细胞克隆技术分离建立不同肺癌细胞亚系NL9980和L9981;应用Southernblot、RT PCR、Westernblot检测NL9980和L9981细胞株中nm 2 3 H1基因多态性、mR NA和蛋白转录表达水平 ;应用MTT法、平板法、Boyden小室法、染色体分型法以及动物实验检测NL9980和L9981细胞株的体内体外生物学特征。结果 ①成功分离建立具有不同转移能力的人大细胞肺癌细胞系NL9980和L9981;②L9981细胞系存在nm 2 3 H1等位基因杂合性缺失、mRNA和蛋白表达缺失 ,而NL9980细胞株nm2 3 H1等位基因结构和转录表达均正常 ;③L9981细胞株的体外增殖能力、克隆形成率和体外侵袭力均显著高于NL9980细胞株 ;④L9981细胞株在裸鼠体内的成瘤性和肺部转移率均显著高于NL9980 ;⑤NL9980和L9981染色体众数比较无显著性差异。结论 ①NL9980和L9981细胞株具有不同的生物学和分子生物学特征 ;②L9981细胞株的高侵袭和高转移潜能可能与nm 2 3 H1等位基因缺失有关。

Objective To explore the possibility of separating and establishing clonal cell subpopulations with different metastatic phenotype from a human lung large cell carcinoma cell line (WCQH-9801), to identify the difference of biological and molecular biology between NL9980 and L9981 cell lines. Methods Two sub-cell lines (NL9980 and L9981) were isolated and established from a human lung large cell carcinoma cell line (WCQH-9801) by the single cell cloning techniques. The RELP, mRNA and protein transcript expression were detected in NL9980 and L9981 cell lines by Southern blot, RT-PCR and Western blot. The biological characteristics of vivo and vitro were determined in NL9980 and L9981 cell lines by MTT, plate, Boyden chamber methods and animal models. Results ①Two sub-cell lines, NL9980 and L9981 which had different metastatic phenotype, were successfully isolated and established from a human lung large cell carcinoma cell line (WCQH-9801). ②The L9981 cell line had LOH of nm23-H1 gene, deletion of mRNA and protein expression of nm23-H1, but the NL9980 cell line had neither LOH of nm23-H1 nor deletion of mRNA and protein expression of nm23-H1. ③The proliferation, clone formation and vitro invastion of L9981 cell line were significantly higher than those of NL9980 cell line. ④The tumorigenicity and lung metastatic rate in nude mouse of L9981 cell line were remarkably higher than those of NL9980. ⑤ No significant difference of the chromosome number was observed between NL9980 and L9981 cell lines. Conclusion ①NL9980 and L9981 cell lines established from a human lung large cell carcinoma cell line have different biological and molecular characteristics. ②The high invsaion and metastasis ability of L9981 cell line might be related to the LOH of nm23-H1 gene.