支气管上皮细胞系BEP2D恶性转化过程中p16基因甲基化改变研究

Study on p16 methylation status of BEP2D cells during its malignant transformation

目的 研究人支气管上皮细胞系BEP2D恶性转化过程中p16基因甲基化改变以及 p16基因mRNA转录情况。方法 选取正常人支气管上皮细胞系BEP2D及其经α粒子照射 2 0周 (R 2 0 )、2 1周 (R 2 1)、3 5周 (T 3 5 )和 5 4周 (T 5 4)的BEP2D细胞株作为研究对象 (其中R 2 0、R 2 1未发生恶性转化 ,而T 3 5、T 5 4已发生恶性转化 ) ,采用甲基化特异性PCR(methylation specificPCR ,MSP)检测各细胞株中 p16基因的甲基化改变情况 ,再运用半定量反转录PCR (retro translationPCR ,RT PCR)检测 p16基因mRNA在各细胞株中的表达情况。结果 ①正常BEP2D细胞株 p16基因未发生甲基化 ,而R 2 0、R 2 1、T 3 5和T 5 4细胞的p16基因均发生了甲基化改变。②与正常BEP2D细胞相比 ,R 2 0、R 2 1、T 3 5和T 5 4细胞 p16基因mRNA转录水平均降低。结论 p16基因甲基化改变发生在肺癌形成的早期阶段。p16基因甲基化可导致

Objective To study p16 methylation status and p16 mRNA transcription of BEP2D cells during its malignant transformation. Methods Normal BEP2D cell and BEP2D cells irradiated by α particle for 20 weeks (R 20), 21 weeks (R 21), 35 weeks (T 35) and 54 weeks (T 54) respectively were chosen to study the p16 methylation status by methylation specific PCR (MSP). Meanwhile, RT PCR was used to study p16 mRNA transcription of the above cells. Results ① p16 methylation was found in R 20, R 21, T 35 and T 54 cells, but not in normal BEP2D cell. ② The p16 mRNA transcription levels of R 20, R 21, T 35 and T 54 cells were much lower than that of normal BEP2D cell. Conclusion The p16 methylation occurs in the early stage of lung cancer. The methylation of p16 gene may cause the inactivation of p16 gene.