云锡矿粉诱导人肺上皮细胞转化和成纤维细胞活化过程中的相互作用  

Interaction between malignant transformation of human pulmonary epithelial cells and activation of fibroblasts induced by Yunnan tin mine dust

作　　者：边莉[1] 何永文[2] 阮永华[2] 唐莹[2] 高倩[2] 王春艳[1] 金克炜[2] 

机构地区：[1]昆明医学院第一附属医院病理科,650031 [2]昆明医学院第一附属医院病理教研室,650031

出　　处：《中华劳动卫生职业病杂志》2009年第7期405-410,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目（30560053）;云南省应用基础研究项目（2006C0042M、2008CD009）;教育部高等学校博士学科点专项科研基金（20050678004）

摘　　要：目的探讨云锡矿粉诱导人肺上皮细胞转化及成纤维细胞活化过程中的相互作用。方法常规培养永生化人支气管上皮细胞（BEAS-2B）及人胚肺成纤维细胞（WI-38），每6天传代1次；100μg／ml的云锡矿粉隔代诱导BEAS-2B及WI-38至第9代，共诱导5次，自第11代分组共培养至第40代。刀豆凝集素A及锚着独立性生长实验检测细胞恶性转化，流式细胞术检测细胞周期变化，免疫细胞化学法检测成纤维细胞α-平滑肌肌动蛋白（α—SMA）表达。结果（1）与正常BEAS-2B细胞相比，矿粉诱导的BEAS-2B细胞单独培养至第28代细胞形态开始出现改变；与WI-38细胞共培养后，细胞形态改变提早至第20代；与矿粉诱导的WI-38细胞共培养后，细胞形态改变进一步提早至第16代。矿粉诱导的BEAS-2B细胞培养至第26代时凝集实验出现阳性；与WI-38细胞及矿粉诱导的WI-38细胞共培养后，细胞凝集时间缩短。与WI-38细胞共培养的矿粉诱导BEAS-2B及与矿粉诱导WI-38细胞共培养的矿粉诱导的BEAS-2B细胞分别在第26及第36代锚着独立性生长实验出现阳性，第36代的克隆形成率分别为6．00‰±1．00‰和15．33‰±2．52‰，且差异有统计学意义（P〈0．05）。矿粉诱导的各组上皮细胞S期细胞所占比例随培养代数增加逐渐升高，细胞发生恶性转化。（2）100μg/ml矿粉诱导的成纤维细胞培养至第26代出现α—SMA表达；与上皮细胞共培养后，成纤维细胞α-SMA表达增强，且随培养代数的增加，阳性表达细胞数明显增多，着色程度增强。结论个旧矿粉能诱导支气管上皮细胞恶性转化及成纤维细胞活化；肺上皮细胞是矿粉诱癌的主要靶细胞；肺上皮细胞的转化和成纤维细胞的活化相互影响，相互促进。Objective To study the interaction between transformation of human pulmonary epithelial cells and activation of fibroblasts induced by Yunnan tin mine dust. Methods （ 1 ） The immortalized human bronchial epithelial cell line BEAS-2B and human embryo lung fibroblast cell line WI-38 were grown in MEM medium containing 5% and 10% FBS, respectively, at 37℃ and 5% CO2 with saturated humidity. The ceils were subcultured every 6 days. BEAS-2B cells and WI-38 cells were induced with Yunnan tin mine dust on every other generation at the concentration of 100 μg/ml. From the 1 lth generation, the cells were co-cultured. Epithelial cell transformation was tested using concanavalin A （ConA）agglutination and anchorage-independent growth assays. The cell cycles were analyzed through flow cytometry. The expressions of α-SMA in fibroblasts were determined with immunocytochemistry. Results （1） Cell morphology of mine dust-exposed epithelial cells began to transform at the 28th generation. Similar transformations were observed with mine dust-induced epithelial cells co-cultured with fibroblasts from the 20th generation and mine dust-induce epithelial cells co-cultured with mine dust-induced fibroblasts from the 16th generation. ConA agglutination assay and anchorage-independent growth assays were negative in normal BEAS-2B ceils. At the 26 th generation, the agglutination test result of the mine dust-exposed epithelial ceils was positive. Co-cultured with fibroblasts and mine dust-exposed fibroblasts, the agglutination time of the mine dust-exposed epithelial cells became short. Epithelial cell anchorage-independent growth assay was positive for mine dust-exposed epithelial cells co-cultured with fibroblasts at the 36th generations and for mine dust-exposed epithelial ceils co-cuhured with mine dust-exposed fibroblasts at the 26th generations. The clone formation rate of the 26th generation was 6.00‰± 1.00‰ and 15.33‰±2.52‰ respectively, with the significant differences （P〈0.05）. With generation addi

关 键 词：云锡矿粉 上皮细胞 成纤维细胞 转化 

分 类 号：R686[医药卫生—骨科学]