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目的 对源自人胚肺组织原代培养、形态似上皮类型的细胞系———人肺上皮细胞(HLEC)的生物学特 性进行鉴定,为相关研究和应用提供模型和依据。 方法 分别采用相差显微镜下观察、细胞计数、β 半乳糖苷酶 染色、免疫荧光细胞化学。人Alu序列PCR扩增、染色体计数、软琼脂集落形成实验和裸鼠致瘤实验对HLEC的形 态学特征、生长特性、衰老状态、上皮细胞标志物、来源、细胞遗传学特点和恶性转化等生物学特性进行研究鉴定。 结果 HLEC角蛋白和上皮型细胞间黏附分子钙黏着素E cadherin免疫荧光染色阳性,染色体众数在41～44,人 Alu序列PCR扩增结果阳性,β 半乳糖苷酶阳性细胞的比例未见随细胞代数的增加而升高,软琼脂和裸鼠体内均不 生长,微丝及黏着斑结构正常。 结论 HLEC细胞具有永生化细胞特征、属相对正常人肺上皮亚二倍体细胞,可 用于肺癌癌变机理研究和癌细胞的相对正常对照。
OBJECTIVE: To identify the biological characteristics of human lung epithelial cells (HLEC), a cell line derived from primary human embryonic lung tissue and morphologically epithelial type, and to provide a model and a basis for related research and application. Methods The cells were observed under phase contrast microscope, cell counting, β-galactosidase staining and immunofluorescence cytochemistry. Human Alu sequence PCR amplification, chromosome count, soft agar colony formation assay and nude mice tumorigenicity test on the morphology, growth characteristics, senescence status, epithelial cell markers, origin, cytogenetic characteristics and malignant transformation of HLEC Study characteristics identified. Results The positive rate of E-cadherin between HLEC keratin and epithelial intercellular adhesion molecule E cadherin was 41 ~ 44, and the positive rate of human Alu sequence was positive. The percentage of β-galactosidase positive cells was not observed Increased cell algebra and increased soft agar and nude mice did not grow in the body, microfilament and focal plaque structure is normal. Conclusions HLEC cells are characterized by immortalized cells. They belong to relatively normal human lung epithelial subdiploid cells and can be used in the study of carcinogenesis of lung cancer and the relative normal control of cancer cells.