食管鳞癌患者PER1、PER2和VEGF表达的相关性及VEGF的节律研究

发布时间：2019-05-12 04:55

【摘要】：目的:测定食管鳞癌患者Period 1(PER1)、Period 2(PER2)和VEGF(vascular endothelial growth factor,VEGF)在食管鳞癌患者中的异常表达与临床病理的关系。了解肿瘤组织中节律基PER1、PER2与节律相关基因VEGF表达的相关性。了解食管鳞癌患者血清、尿液中VEGF的波动规律及其与正常对照人群血液中VEGF波动差异。方法:(1)RT-PCR法检测3例食管鳞癌患者的正常组织、癌旁非典型增生组织及癌组中PER1、PER2和VEGF mRNA表达水平。(2)、Western-Blot法检测食管鳞癌患者的正常组织、癌旁非典型增生组织及癌组织中VEGF、PER1和PER2蛋白表达水平。(3)、收集144例食管鳞癌患者的癌组织、癌旁组织及正常组织、患者血液标本及临床病例特点。免疫组化方法检测144例食管鳞癌患者的癌组织、癌旁组织及正常组织中PER1、PER2和VEGF蛋白表达水平并观察PER、PER2和VEGF与临床特征的相关性。(4)、ELISA方法检测144例患者血液中VEGF的表达情况。(5)、收集9位食管鳞癌患者四个时间点(03:00,09:00,15:00,21:00)血液和尿液标本以及9位正常对照四个时间点(03:00,09:00,15:00,21:00)血液标本。ELISA方法检测9位食管鳞癌患者四个时间点(03:00,09:00,15:00,21:00)血液和尿液标本中VEGF的表达量以及9位正常人血液中VEGF的表达量了解VEGF节律波动的规律。结果:(1)RT-PCR、WB检测结果显示VEGF、PER1和PER2在食管鳞癌患者癌组织、癌旁非典型增生组织及正常组织中的表达存在差异,VEGF在癌组织、癌旁非典型增生组织中表达较正常组织高,PER1和PER2在肿瘤组织、癌旁非典型增生组织中表达较正常组织低。(2)、PER1、PER2蛋白在食管鳞癌患者肿瘤组织中的表达与肿瘤组织的的临床分期、浸润深度、淋巴结转移、远处转移成负相关,与分化程度呈正相关;同时也观察到肿瘤组织越靠近食管下段,其PER1、PER2的表达越低;而与性别、年龄、文化程度、肿瘤大小不相关。肿瘤组织中、血液中VEGF蛋白的表达与肿瘤组织的大小、临床分期、浸润深度、淋巴结转移、远处转移成正相关,与分化程度呈负相关;同时也观察到肿瘤组织越靠近食管下段,其VEGF蛋白的表达越低。而与性别、年龄、文化程度不相关。肿瘤组织中PER1、PER2蛋白的表达与肿瘤组织中、血液中VEGF蛋白表达呈负相关。(3)、正常人血液与食管鳞癌患者尿液、血液中VEGF的表达呈日节律波动,且正常人与食管鳞癌患者一天四个时间点VEGF的波动幅度存在差异。结论:(1)、节律基因PER1和PER2的低表达与VEGF的高表达在食管鳞癌的发生与发展中起了重要作用。(2)、节律基因PER1和PER2的表达水平影响了VEGF表达及波动情况。
[Abstract]:Aim: to investigate the relationship between the abnormal expression of Period 1 (PER1), Period 2 (PER2) and VEGF (vascular endothelial growth factor,VEGF) and clinicopathology in patients with esophageal squamous cell carcinoma. To investigate the relationship between rhythmic PER1,PER2 and rhythm related gene VEGF expression in tumor tissues. To investigate the fluctuation of VEGF in serum and urine of patients with esophageal squamous cell carcinoma and the difference of VEGF in blood between patients with esophageal squamous cell carcinoma and normal controls. Methods: (1) the expression of PER1,PER2 and VEGF mRNA in normal tissues, paracancerous atypical hyperplasia tissues and cancer groups were detected by RT-PCR. (2) the normal tissues of patients with esophageal squamous cell carcinoma were detected by Western-Blot. The expression levels of VEGF,PER1 and PER2 protein in paracancerous atypical hyperplasia and cancer tissues were collected. (3) the characteristics of cancer tissues, paracancerous tissues and normal tissues, blood samples and clinical cases were collected. The expression of PER1,PER2 and VEGF proteins in cancer tissues, paracancerous tissues and normal tissues of 144 patients with esophageal squamous cell carcinoma were detected by immunohistochemical method, and the correlation between PER,PER2 and VEGF and clinical features was observed. (4), The expression of VEGF in blood of 144 patients was detected by ELISA. (5), Blood and urine samples were collected from 9 patients with esophageal squamous cell carcinoma at four time points (0.3, 09, 15, 21) and 9 normal controls at four time points (0.3, 09, 15, 21). Elisa was used to detect 9 food. The expression of VEGF in blood and urine samples and the expression of VEGF in 9 normal subjects at four time points (03: 00, 09: 00, 15: 00, 21: 00) in patients with tubular squamous cell carcinoma understood the fluctuation of VEGF rhythm. Results: (1) the results of RT-PCR,WB showed that there were differences in the expression of VEGF,PER1 and PER2 in cancer tissues, atypical hyperplasia tissues and normal tissues, and the expression of VEGF was different in cancer tissues. The expression of PER1 and PER2 in paracancerous atypical hyperplasia tissues was higher than that in normal tissues. (2) PER1, was lower in tumor tissues and paracancerous atypical hyperplasia tissues than in normal tissues. The expression of PER2 protein in esophageal squamous cell carcinoma was negatively correlated with the clinical stage, depth of invasion, lymph node metastasis and distant metastasis, and positively correlated with the degree of differentiation. At the same time, it was also observed that the closer the tumor tissue was to the lower esophageal segment, the lower the expression of PER1,PER2 was, but not related to sex, age, educational level and tumor size. In tumor tissues, the expression of VEGF protein in blood was positively correlated with the size of tumor tissue, clinical stage, depth of invasion, lymph node metastasis and distant metastasis, but negatively correlated with the degree of differentiation. At the same time, the closer the tumor tissue was to the lower esophageal segment, the lower the expression of VEGF protein was. It is not related to sex, age and educational level. The expression of PER1,PER2 protein in tumor tissue was negatively correlated with the expression of VEGF protein in blood. (3) the expression of VEGF in urine and blood of normal human blood and esophageal squamous cell carcinoma patients fluctuated in daily rhythm. There was significant difference in the fluctuation range of VEGF between normal subjects and patients with esophageal squamous cell carcinoma at four time points a day. Conclusion: (1) the low expression of rhythm genes PER1 and PER2 and the high expression of VEGF play an important role in the occurrence and development of esophageal squamous cell carcinoma. (2) the expression of rhythm genes PER1 and PER2 affects the expression and fluctuation of VEGF.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.1

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