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目的研究温石棉诱导人支气管上皮细胞系的BEAS-2B细胞恶性转化活力及错配修复基因表达的变化。方法以2×10-3μg/m2温石棉纤维染毒BEAS-2B细胞,每10 d染毒24 h,待细胞出现生长抑制后用锚着不依赖性生长实验判断细胞恶性转化,并利用实时定量逆转录聚合酶链反应(qRT-PCR)方法检测温石棉对BEAS-2B染毒12 h、24 h、48 h、60 d等不同染毒阶段的人类错配修复基因hMSH1和hMLH2的mRNA表达的变化。结果细胞染毒60 d后生长加快,接触抑制消失,呈现重叠生长,并可在软琼脂中生长。温石棉染毒24 h和48 h组mRNA表达下调,60 d组表达上调。结论温石棉可导致人支气管上皮细胞BEAS-2B错配修复基因表达发生变化。
Objective To study the changes of malignant transformation activity and mismatch repair gene expression of BEAS-2B cells induced by chrysotile in human bronchial epithelial cell line. Methods BEAS-2B cells were exposed to 2 × 10-3μg / m2 chrysotile asbestos fibers for 24 hours every 10 days. When the cells appeared growth inhibition, the malignant transformation of cells was judged by the anchorage independent growth experiment. The real-time quantitative Reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect mRNA expression of human mismatch repair genes hMSH1 and hMLH2 in different stages of exposure to BEAS-2B at 12 h, 24 h, 48 h, 60 d Variety. Results The cells grew rapidly after 60 days of exposure and disappeared after contact inhibition. The cells grew overlappingly and grew on soft agar. In chrysotile exposed to chrysotile at 24 h and 48 h, the mRNA expression was down-regulated and up-regulated at 60 d. Conclusions Chrysotile can change the expression of BEAS-2B mismatch repair gene in human bronchial epithelial cells.