时钟基因Per1、Per2、Per3在头颈部鳞癌组织中的表达及与临床相关因素的研究

发布时间：2018-03-09 07:13

本文选题：头颈部鳞癌　切入点：头颈部正常组织　出处：《贵州医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:从蛋白、基因水平研究时钟基因Per1、Per2、Per3在头颈部鳞癌及正常组织中的表达与临床特征、分期和生存之间的联系,望为临床评估患者疗效及预后提供参考依据。方法:收集头颈部鳞癌组织标本20例,头颈部正常组织标本7例,采用实时荧光定量PCR法检测Per1、Per2、Per3基因的表达水平,通过相对定量法分析Per1、Per2、Per3基因在头颈部鳞癌组织及头颈部正常组织中的表达是否有差异,及头颈部鳞癌组织中Per1、Per2、Per3基因表达水平与早期、晚期患者之间的关系。同时采用免疫组化法检测60例头颈部鳞癌标本及20例头颈部正常组织标本中Per1、Per2、Per3蛋白表达情况,运用染色程度分级与染色细胞百分比相乘结果测定Per1、Per2、Per3蛋白表达水平,分析头颈部鳞癌组织及头颈部正常组织中Per1、Per2、Per3蛋白之间的关系,并从蛋白水平分析其与患者临床特征、肿瘤临床分期、即刻疗效和生存之间的关系。结果:实时荧光定量PCR法检测显示Per1、Per2、Per3基因在头颈部鳞癌组织中及头颈部正常组织中均有表达,但头颈部鳞癌组织中Per1、Per2、Per3基因的表达较正常组织明显下降(P0.01),晚期患者中Per1、Per2、Per3基因下调水平均高于早期患者,且Per2、Per3基因下调程度明显高于早期患者,差异有统计学意义(P0.01)。运用免疫组化法检测表明头颈部鳞癌组织中Per1、Per2、Per3蛋白阳性表达率明显低于正常组织中的阳性表达率,差异有统计学意义(P0.05),且头颈部鳞癌组织中Per1、Per2、Per3蛋白阳性表达与肿瘤临床分期有明显相关性(P0.05),但其与患者年龄、性别无关。即刻疗效方面,Per1、Per2、Per3蛋白阳性组患者其治疗有效率高于阴性组患者,其结果具有统计学意义(P0.05)。头颈部鳞癌患者中Per1、Per2、Per3蛋白阳性组患者的1、2、3年OS明显高于阴性组,差异有统计学意义(P0.05);Per1、Per2、Per3蛋白阳性组患者的1、2、3年PFS、RFS均明显高于阴性组,差异有统计学意义(P0.05)。结论:Per家族基因表达的降低可能与头颈部鳞癌的发生具有相关性,且在晚期患者中其Per2、Per3基因的下调程度高于早期头颈部鳞癌患者;Per家族蛋白表达阳性的头颈部鳞癌患者较阴性患者其可能具有更长的OS及PFS,RFS,而这也提示Per家族基因与头颈部鳞癌的预后中具有重要作用。
[Abstract]:Objective: to study the relationship between the expression of clock gene Per1, Per2P3 and clinical features, staging and survival in head and neck squamous cell carcinoma and normal tissues at the protein and gene levels. Methods: 20 cases of head and neck squamous cell carcinoma and 7 cases of normal head and neck tissues were collected to detect the expression of Per1 PER2P 3 gene by real-time fluorescence quantitative PCR. The relative quantitative analysis was used to analyze the difference of the expression of Per1mPer2P3 gene in head and neck squamous cell carcinoma and normal head and neck tissues, and the expression level and early stage in head and neck squamous cell carcinoma. Immunohistochemical method was used to detect the expression of Per1 and Per2P3 protein in 60 cases of head and neck squamous cell carcinoma and 20 cases of normal tissues of head and neck. The expression level of Per1 PER2P 3 protein in head and neck squamous cell carcinoma and normal tissue of head and neck were determined by staining grade and percentage of stained cells. The relationship between Per1 and Per2 Per3 protein in head and neck squamous cell carcinoma and normal tissue of head and neck was analyzed, and the clinical features of patients were analyzed from the protein level. Results: real time fluorescence quantitative PCR assay showed that the expression of Per1mPer2p3 gene was detected in head and neck squamous cell carcinoma tissues and normal head and neck tissues. But in head and neck squamous cell carcinoma, the expression of Per1, Per2, P0. 01, P0. 01, and the down-regulation level of Per1, Per2, Per2, Per3, and the down-regulation of Per2, P0. 01, and the down-regulation of Per2, P0. 01, were significantly lower than those in the early stage of squamous cell carcinoma. The difference was statistically significant (P 0.01). The positive expression rate of Per1, Per2, Per3 protein in head and neck squamous cell carcinoma tissues was significantly lower than that in normal tissues, and the positive expression rate in head and neck squamous cell carcinoma tissues was significantly lower than that in normal tissues. The difference was statistically significant (P 0.05), and the positive expression of Per1 / Per2 / Per3 protein in head and neck squamous cell carcinoma tissues was significantly correlated with the clinical stage of the tumor (P 0.05), but it was associated with the age of the patients. The effective rate of treatment was higher in the positive group than that in the negative group, and the results were statistically significant (P 0.05). In the patients with head and neck squamous cell carcinoma, the results were significantly higher than those in the group with positive protein of Per1P 2 and P 2 + 3, and the OS in 3 years was significantly higher than that in the negative group. The difference was statistically significant (P 0.05) in the patients with positive P0. 05% Per1 and Per2 + Per3 protein, and in 3 years, the RFS of PFSn was significantly higher than that of the negative group, and the difference was statistically significant (P 0. 05). Conclusion the decrease of the gene expression of the weight per family may be related to the occurrence of squamous cell carcinoma of the head and neck. Moreover, the down-regulation of Per2P3 gene in advanced patients was higher than that in patients with head and neck squamous cell carcinoma with positive protein expression of per family. It may have longer OS and PFS- FRFSs in patients with head and neck squamous cell carcinoma than in patients with negative expression. This also suggests that Per family genes are more likely to be present in patients with squamous cell carcinoma of head and neck than those in patients with negative expression of per family proteins. It plays an important role in prognosis of head and neck squamous cell carcinoma.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.91

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