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目的检测锯齿状病变组织中p16基因启动子区异常甲基化改变和p16蛋白的表达情况,探讨其在锯齿状癌变通路中的作用和临床病理意义,同时探讨p16基因在不同年龄层段甲基化程度。方法采用Taqman探针实时定量PCR(Methylight)方法检测225例锯齿状病变(包括96例HP、61例SSA/P和68例TSA)、54例TA、69例CRC和42例正常结直肠黏膜组织中的p16基因甲基化状态,并通过测序法验证扩增的目的片段;同时应用免疫组化方法检测其中随机抽取的116例锯齿状病变(包括52例HP、41例SSA/P、23例TSA)、20例TA、24例CRC和24例正常结直肠黏膜组织中p16蛋白的表达。结果 p16基因启动子甲基化状态和p16蛋白异常阳性程度在锯齿状病变和对照组中均有显著差异(P<0.05),两者比较呈负相关(P<0.05);p16基因启动子甲基化频率与不同年龄层段的相关性比较有显著差异(P<0.05),两者呈正相关。结论结直肠锯齿状病变组织中p16基因甲基化可能是诱导其蛋白表达下调的主要原因,且在结直肠“增生性息肉-锯齿状腺瘤-癌”的锯齿状癌变通路中起重要作用;同时p16基因甲基化又可能与年龄相关,随年龄增长甲基化程度越高。
Objective To detect the abnormal methylation of p16 gene and the expression of p16 protein in serrated lesions and to explore the role of p16 gene in clinicopathological significance of serrated lesions and to explore the role of p16 gene in the methylation of different age groups . Methods 225 cases of serrated lesions (including 96 cases of HP, 61 cases of SSA / P and 68 cases of TSA), 54 cases of TA, 69 cases of CRC and 42 cases of normal colorectal mucosa were detected by Taqman real-time quantitative PCR (Methylight) The p16 gene was methylated and the amplified fragment was verified by sequencing. 116 randomly selected serrated lesions (including 52 HP, 41 SSA / P, 23 cases TSA), 20 cases of TA, 24 cases of CRC and 24 cases of normal colorectal mucosa tissue p16 protein expression. Results The methylation status of p16 gene and the positive expression of p16 protein were significantly different between the serrated lesions and the control group (P <0.05), and negative correlation was found between them (P <0.05). The promoter of p16 gene There was a significant difference in the correlation between the frequency of alkalization and different age groups (P <0.05), and there was a positive correlation between them. Conclusion Methylation of p16 gene in the serrated lesions of the colorectum may be the main reason for the down-regulation of its protein expression and plays an important role in the serration of the serrated lesions of the colorectal polyps - serrated adenoma - carcinoma Role; at the same time p16 gene methylation may be age-related, with age, the higher the degree of methylation.