本文選題：N-myc + 下游調(diào)節(jié)基因1��； 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：目的觀察人類N-myc下游調(diào)節(jié)基因1（NDRG1）、COX-2及VEGF在宮頸鱗癌組織中的表達及意義；建立穩(wěn)定轉(zhuǎn)染含NDRG1的宮頸癌SiHa細胞株,探討NDRG1對SiHa細胞增殖與凋亡的影響及其可能的分子機制，為宮頸鱗癌的診治和判斷預(yù)后提供依據(jù)。 方法應(yīng)用免疫組織化學(xué)方法檢測15例正常宮頸組織和80例宮頸癌組織中NDRG1、COX-2及VEGF蛋白的表達情況，，并分析其表達與臨床病理因素及患者預(yù)后的關(guān)系；運用Spearman秩相關(guān)觀察三者蛋白表達的相關(guān)性；運用Kaplan-Meier生存曲線分析其表達與患者預(yù)后的關(guān)系。經(jīng)脂質(zhì)體介導(dǎo)將含有NDRG1真核表達質(zhì)粒轉(zhuǎn)染人宮頸癌SiHa細胞株，采用G418篩選陽性細胞克隆及逆轉(zhuǎn)錄聚合酶鏈式反應(yīng)法（RT-PCR）、Western blot；RT-PCR和Western blot檢測陽性細胞克隆COX-2及VEGF表達的改變；噻唑鹽（MTT）比色法檢測穩(wěn)定高表達NDRG1的陽性細胞克隆N12的增殖活性；流式細胞儀檢測陽性細胞克隆N12的凋亡率。 結(jié)果（1）80例宮頸鱗癌組織中NDRG1陽性表達率為52.2%，顯著低于正常宮頸組織中表達水平（P0.01）；COX-2與VEGF陽性表達率分別為91.2%及77.5%，顯著高于正常宮頸組織中表達水平（P0.01）；NDRG1、COX-2及VEGF蛋白的陽性表達與宮頸鱗癌患者的年齡及腫瘤大小無關(guān)（P0.05），而與腫瘤組織病理學(xué)分級、FIGO分期及淋巴結(jié)轉(zhuǎn)移密切相關(guān)（P0.01）。（2）Kaplan-Meier生存分析顯示，宮頸鱗癌組織中NDRG1表達陽性者生存期較長，COX-2及VEGF表達陽性者生存期較短。 結(jié)論(1)宮頸鱗癌組織中NDRG1表達降低，而COX-2及VEGF表達增加，聯(lián)合檢測宮頸鱗癌組織中NDRG1、COX-2及VEGF的表達可能對判斷預(yù)后具有重要價值。（2）上調(diào)SiHa細胞NDRG1表達后，細胞凋亡增加而細胞增殖降低，其作用機制可能與COX-2、VEGF表達下降有關(guān)。
[Abstract]:Objective to investigate the expression and significance of COX-2 and VEGF in cervical squamous cell carcinoma (SCC), and to investigate the effect of NDRG1 on the proliferation and apoptosis of SiHa cells and its possible molecular mechanism. To provide a basis for the diagnosis, treatment and prognosis of cervical squamous cell carcinoma. Methods Immunohistochemical method was used to detect the expression of NDRG1COX-2 and VEGF protein in 15 normal cervical tissues and 80 cervical carcinoma tissues, and the relationship between the expression and clinicopathological factors and prognosis of the patients was analyzed. Spearman rank correlation was used to observe the correlation of protein expression and Kaplan-Meier survival curve was used to analyze the relationship between the expression and the prognosis of the patients. The eukaryotic expression plasmid containing NDRG1 was transfected into human cervical cancer SiHa cell line mediated by liposome. The positive cells were screened by G418 and reverse transcriptase polymerase chain reaction was used to detect the expression of COX-2 and VEGF by RT-PCR and Western blot. The proliferative activity of positive cell clone N12 with stable and high expression of NDRG1 was detected by MTT colorimetry, and the apoptosis rate of positive cell clone N12 was detected by flow cytometry. Results the positive expression rate of NDRG1 in 80 cases of cervical squamous cell carcinoma was 52.2, which was significantly lower than that in normal cervical tissues. The positive rates of COX-2 and VEGF were 91.2% and 77.5%, respectively, which were significantly higher than those in normal cervical tissues. Sex expression was not correlated with age and tumor size of cervical squamous cell carcinoma patients, but was closely correlated with Figo stage of tumor histopathology and lymph node metastasis. Kaplan-Meier survival analysis showed that The survival time of patients with positive NDRG1 expression was longer than that of patients with positive expression of COX-2 and VEGF. Conclusion (1) the expression of NDRG1 in cervical squamous cell carcinoma is decreased, while the expression of COX-2 and VEGF is increased. The combined detection of COX-2 and VEGF in cervical squamous cell carcinoma may play an important role in judging the prognosis. The increase of apoptosis and the decrease of cell proliferation may be related to the decrease of VEGF expression in COX-2.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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