【摘要】：目的:測定食管鱗癌患者Period 1(PER1)、Period 2(PER2)和VEGF(vascular endothelial growth factor,VEGF)在食管鱗癌患者中的異常表達與臨床病理的關(guān)系。了解腫瘤組織中節(jié)律基PER1、PER2與節(jié)律相關(guān)基因VEGF表達的相關(guān)性。了解食管鱗癌患者血清、尿液中VEGF的波動規(guī)律及其與正常對照人群血液中VEGF波動差異。方法:(1)RT-PCR法檢測3例食管鱗癌患者的正常組織、癌旁非典型增生組織及癌組中PER1、PER2和VEGF mRNA表達水平。(2)、Western-Blot法檢測食管鱗癌患者的正常組織、癌旁非典型增生組織及癌組織中VEGF、PER1和PER2蛋白表達水平。(3)、收集144例食管鱗癌患者的癌組織、癌旁組織及正常組織、患者血液標本及臨床病例特點。免疫組化方法檢測144例食管鱗癌患者的癌組織、癌旁組織及正常組織中PER1、PER2和VEGF蛋白表達水平并觀察PER、PER2和VEGF與臨床特征的相關(guān)性。(4)、ELISA方法檢測144例患者血液中VEGF的表達情況。(5)、收集9位食管鱗癌患者四個時間點(03:00,09:00,15:00,21:00)血液和尿液標本以及9位正常對照四個時間點(03:00,09:00,15:00,21:00)血液標本。ELISA方法檢測9位食管鱗癌患者四個時間點(03:00,09:00,15:00,21:00)血液和尿液標本中VEGF的表達量以及9位正常人血液中VEGF的表達量了解VEGF節(jié)律波動的規(guī)律。結(jié)果:(1)RT-PCR、WB檢測結(jié)果顯示VEGF、PER1和PER2在食管鱗癌患者癌組織、癌旁非典型增生組織及正常組織中的表達存在差異,VEGF在癌組織、癌旁非典型增生組織中表達較正常組織高,PER1和PER2在腫瘤組織、癌旁非典型增生組織中表達較正常組織低。(2)、PER1、PER2蛋白在食管鱗癌患者腫瘤組織中的表達與腫瘤組織的的臨床分期、浸潤深度、淋巴結(jié)轉(zhuǎn)移、遠處轉(zhuǎn)移成負相關(guān),與分化程度呈正相關(guān);同時也觀察到腫瘤組織越靠近食管下段,其PER1、PER2的表達越低;而與性別、年齡、文化程度、腫瘤大小不相關(guān)。腫瘤組織中、血液中VEGF蛋白的表達與腫瘤組織的大小、臨床分期、浸潤深度、淋巴結(jié)轉(zhuǎn)移、遠處轉(zhuǎn)移成正相關(guān),與分化程度呈負相關(guān);同時也觀察到腫瘤組織越靠近食管下段,其VEGF蛋白的表達越低。而與性別、年齡、文化程度不相關(guān)。腫瘤組織中PER1、PER2蛋白的表達與腫瘤組織中、血液中VEGF蛋白表達呈負相關(guān)。(3)、正常人血液與食管鱗癌患者尿液、血液中VEGF的表達呈日節(jié)律波動,且正常人與食管鱗癌患者一天四個時間點VEGF的波動幅度存在差異。結(jié)論:(1)、節(jié)律基因PER1和PER2的低表達與VEGF的高表達在食管鱗癌的發(fā)生與發(fā)展中起了重要作用。(2)、節(jié)律基因PER1和PER2的表達水平影響了VEGF表達及波動情況。
[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.
【學位授予單位】：川北醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R735.1

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