Zhou Qinghua, Wang Yanping, Che Guowei, Zhu Wen, Chen Xiaohe, Chen Xiaofeng, Sun Zhilin
Key Laboratory of Lung Cancer Molecular Biology of Sichuan Province, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R.China.
Zhongguo Fei Ai Za Zhi. 2003 Dec 20;6(6):464-8. doi: 10.3779/j.issn.1009-3419.2003.06.13.
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.
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.
(1)Two 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). (2)The 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. (3)The proliferation, clone formation and vitro invastion of L9981 cell line were significantly higher than those of NL9980 cell line. (4)The 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.
(1)NL9980 and L9981 cell lines established from a human lung large cell carcinoma cell line have different biological and molecular characteristics. (2)The high invsaion and metastasis ability of L9981 cell line might be related to the LOH of nm23-H1 gene.
探讨从人肺大细胞癌细胞系（WCQH-9801）中分离并建立具有不同转移表型的克隆细胞亚群的可能性，以明确NL9980和L9981细胞系在生物学及分子生物学方面的差异。
采用单细胞克隆技术从人肺大细胞癌细胞系（WCQH-9801）中分离并建立了两个亚细胞系（NL9980和L9981）。通过Southern印迹、RT-PCR和Western印迹检测NL9980和L9981细胞系中的RELP、mRNA和蛋白转录表达。采用MTT、平板、Boyden小室法及动物模型检测NL9980和L9981细胞系的体内外生物学特性。
（1）成功从人肺大细胞癌细胞系（WCQH-9801）中分离并建立了具有不同转移表型的两个亚细胞系NL9980和L9981。（2）L9981细胞系存在nm23-H1基因杂合性缺失，nm23-H1的mRNA及蛋白表达缺失，而NL9980细胞系既无nm23-H1基因杂合性缺失，也无nm23-H1的mRNA及蛋白表达缺失。（3）L9981细胞系的增殖、克隆形成及体外侵袭能力明显高于NL9980细胞系。（4）L9981细胞系在裸鼠中的致瘤性及肺转移率显著高于NL9980。（5）NL9980和L9981细胞系之间的染色体数目未观察到显著差异。
（1）从人肺大细胞癌细胞系建立的NL9980和L9981细胞系具有不同的生物学及分子特征。（2）L9981细胞系的高侵袭和转移能力可能与nm23-H1基因的杂合性缺失有关。
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