鼻咽癌中PCDH8基因甲基化的研究

发布时间：2018-04-18 17:30

本文选题：PCDH8 + 甲基化　；参考：《重庆医科大学》2012年硕士论文

【摘要】：目的：PCDH8（Protocadherin8）基因是部分肿瘤发展进程中的一种肿瘤抑制物，本实验旨在研究PCDH8的甲基化水平及其在鼻咽癌中的抑癌功能。方法：采用逆转录聚合酶链反应（RT-PCR），检测鼻咽癌细胞系C666-1、CNE1、HK1、HNE1、HONE1，正常的人的鼻咽上皮细胞系NP69及下咽癌细胞系Fadu里面PCDH8mRNA的转录的水平。在细胞系、41例鼻咽癌组织和16例慢性鼻咽炎组织中PCDH8启动子的甲基化水平用甲基化特异性PCR（MSP）来进行测定。然后再用该基因转染鼻咽癌细胞CNE1，检测该基因对该细胞的生理作用。结果：在正常鼻咽上皮细胞系NP69及下咽癌细胞系Fadu里面，PCDH8基因是稳定表达的。在这些鼻咽癌细胞系里面，HK1和HNE1细胞的表达是下调的，另三株均的表达是缺失的。甲基化的分析显示，PCDH8基因在鼻咽癌细胞系及Fadu细胞系中均发生了甲基化，，鼻咽癌组织中甲基化率为85.3%(35/41)，而在正常鼻咽细胞系NP69和慢性鼻咽炎组织中均未发生甲基化。PCDH8基因的启动子的甲基化的水平与鼻咽癌病人的临床的病理参数全部没有任何的联系。用5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycytidine，Aza)和曲古菌素A （trichostatin A，TSA）处理鼻咽癌细胞后，可恢复PCDH8的表达。PCDH8在鼻咽癌细胞里面的过表达，抑制了CNE1的生长和移动。结论：本实验第一次研究说明了PCDH8基因在鼻咽癌的细胞和组织里面都发生了启动子的甲基化，它还可以抑制CNE1的生长和移动。所以，PCDH8基因可以作为鼻咽癌的一个候选的抑瘤基因。
[Abstract]:Objective to investigate the methylation level of PCDH8 and its function in nasopharyngeal carcinoma (NPC).Methods: reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the transcription of PCDH8mRNA in nasopharyngeal carcinoma cell line C666-1, normal human nasopharyngeal epithelial cell line NP69 and hypopharyngeal carcinoma cell line Fadu.The methylation level of PCDH8 promoter in 41 nasopharyngeal carcinoma tissues and 16 chronic nasopharyngitis tissues was determined by methylation-specific PCDH8 MSPs.Then the gene was transfected into nasopharyngeal carcinoma cell line CNE 1, and the physiological effect of the gene on CNE 1 cells was detected.Results: in normal nasopharyngeal epithelial cell line NP69 and hypopharyngeal carcinoma cell line Fadu, the expression of pPCDH8 gene was stable.In these nasopharyngeal carcinoma cell lines, the expression of HK1 and HNE1 cells was down-regulated and the other three strains were absent.Methylation analysis showed that the PCDH8 gene was methylated in both nasopharyngeal carcinoma and Fadu cell lines.The methylation rate in nasopharyngeal carcinoma tissues was 85.3% and 35 / 41%, while no methylation level of promoter of methyl.PCDH8 gene was found in normal nasopharyngeal cell line NP69 and chronic nasopharyngitis tissue. There was no correlation between the methylation level and the clinicopathological parameters of nasopharyngeal carcinoma patients.After treated with 5-aza-2-deoxycytidine (Aza) and trichostatin (TSA), the overexpression of PCDH8. PCDH8 in nasopharyngeal carcinoma cells was restored, which inhibited the growth and migration of CNE1.Conclusion: this first study demonstrated that the promoter methylation of PCDH8 gene occurs in nasopharyngeal carcinoma cells and tissues, and it can also inhibit the growth and migration of CNE1.Therefore, PCDH 8 gene can be used as a candidate tumor suppressor gene for nasopharyngeal carcinoma.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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