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(guān)察人類(lèi)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(dá)及意義；建立穩(wěn)定轉(zhuǎn)染含NDRG1的宮頸癌SiHa細(xì)胞株,探討NDRG1對(duì)SiHa細(xì)胞增殖與凋亡的影響及其可能的分子機(jī)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(cè)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(dá)情況，，并分析其表達(dá)與臨床病理因素及患者預(yù)后的關(guān)系；運(yùn)用Spearman秩相關(guān)觀(guān)察三者蛋白表達(dá)的相關(guān)性；運(yùn)用Kaplan-Meier生存曲線(xiàn)分析其表達(dá)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(dá)質(zhì)粒轉(zhuǎn)染人宮頸癌SiHa細(xì)胞株，采用G418篩選陽(yáng)性細(xì)胞克隆及逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)法（RT-PCR）、Western blot；RT-PCR和Western blot檢測(cè)陽(yáng)性細(xì)胞克隆COX-2及VEGF表達(dá)的改變；噻唑鹽（MTT）比色法檢測(cè)穩(wěn)定高表達(dá)NDRG1的陽(yáng)性細(xì)胞克隆N12的增殖活性；流式細(xì)胞儀檢測(cè)陽(yáng)性細(xì)胞克隆N12的凋亡率。 結(jié)果（1）80例宮頸鱗癌組織中NDRG1陽(yáng)性表達(dá)率為52.2%，顯著低于正常宮頸組織中表達(dá)水平（P0.01）；COX-2與VEGF陽(yáng)性表達(dá)率分別為91.2%及77.5%，顯著高于正常宮頸組織中表達(dá)水平（P0.01）；NDRG1、COX-2及VEGF蛋白的陽(yáng)性表達(dá)與宮頸鱗癌患者的年齡及腫瘤大小無(wú)關(guān)（P0.05），而與腫瘤組織病理學(xué)分級(jí)、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(dá)陽(yáng)性者生存期較長(zhǎng)，COX-2及VEGF表達(dá)陽(yáng)性者生存期較短。 結(jié)論(1)宮頸鱗癌組織中NDRG1表達(dá)降低，而COX-2及VEGF表達(dá)增加，聯(lián)合檢測(cè)宮頸鱗癌組織中NDRG1、COX-2及VEGF的表達(dá)可能對(duì)判斷預(yù)后具有重要價(jià)值。（2）上調(diào)SiHa細(xì)胞NDRG1表達(dá)后，細(xì)胞凋亡增加而細(xì)胞增殖降低，其作用機(jī)制可能與COX-2、VEGF表達(dá)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(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R737.33

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