GMA致人支气管上皮细胞恶性转化过程中抑癌基因P15甲基化状态的研究

Study on the anti-oncogene of P15 methylation status in malignant transformation of human bronchial epithelial cells induced by glycidyl methacrylate

目的:观察甲基丙烯酸环氧丙酯(glycidyl methacrylate,GMA)致人支气管上皮(16HBE)细胞恶性转化过程中不同时点P15基因甲基化状态,探讨其在细胞恶性转化过程中可能的作用。方法:收获GMA转化前期(第10代)、转化中期(第20代)、转化后期(第30代)16HBE细胞,应用甲基化芯片技术和甲基化特异性PCR(MSP)检测P15基因在GMA诱导16HBE细胞恶性转化不同阶段的甲基化状态。结果:甲基化芯片结果显示,P15基因在转化中期、转化后期出现甲基化而转化前期未见甲基化,MSP法检测结果与芯片结果一致。结论:P15基因可能为GMA诱导细胞恶性程度相关的特异性基因,可考虑将其作为诱导细胞恶性转化的生物标志物。

OBJECTIVE： Analyzing the changes of P15 methylation status among different stages of malignant transformation of human bronchial epithelial cells （16HBE） induced by glycidyl methacrylate （GMA） and discussing the role of DNA methylation in the process. METHODS： Cells were harvested at different times, protophase （the 10th generation）, metaphase （the 20th generation） and anaphase （the 30th generation） of transformed cells. Then these methylation statuses of P15 were detected by methylation chip and methylation-specific PCR （MSP）, and compared to the control groups （treated with DMSO） of the same generation. RESULTS： Based on the result of methylation chip and MSP, the methylation of P15 gene in 16HBE cells transformed by GMA occurred in both metaphase and anaphase but not in protophase. CONCLUSION： Gene of P15 could be regarded as a specific gene related to the degree of malignancy and a particular mutation biomarker of cell malignant transformation. [KEY WORDS1 glycidyl methacrylate; human bronchial epithelial cells ; malignant transformation; methylation