NDRG1基因在宫颈癌细胞及组织中的表达及意义

发布时间：2018-05-05 05:29

本文选题：N-myc + 下游调节基因1　；参考：《苏州大学》2014年硕士论文

【摘要】：目的观察人类N-myc下游调节基因1（NDRG1）、COX-2及VEGF在宫颈鳞癌组织中的表达及意义；建立稳定转染含NDRG1的宫颈癌SiHa细胞株,探讨NDRG1对SiHa细胞增殖与凋亡的影响及其可能的分子机制，为宫颈鳞癌的诊治和判断预后提供依据。方法应用免疫组织化学方法检测15例正常宫颈组织和80例宫颈癌组织中NDRG1、COX-2及VEGF蛋白的表达情况，，并分析其表达与临床病理因素及患者预后的关系；运用Spearman秩相关观察三者蛋白表达的相关性；运用Kaplan-Meier生存曲线分析其表达与患者预后的关系。经脂质体介导将含有NDRG1真核表达质粒转染人宫颈癌SiHa细胞株，采用G418筛选阳性细胞克隆及逆转录聚合酶链式反应法（RT-PCR）、Western blot；RT-PCR和Western blot检测阳性细胞克隆COX-2及VEGF表达的改变；噻唑盐（MTT）比色法检测稳定高表达NDRG1的阳性细胞克隆N12的增殖活性；流式细胞仪检测阳性细胞克隆N12的凋亡率。结果（1）80例宫颈鳞癌组织中NDRG1阳性表达率为52.2%，显著低于正常宫颈组织中表达水平（P0.01）；COX-2与VEGF阳性表达率分别为91.2%及77.5%，显著高于正常宫颈组织中表达水平（P0.01）；NDRG1、COX-2及VEGF蛋白的阳性表达与宫颈鳞癌患者的年龄及肿瘤大小无关（P0.05），而与肿瘤组织病理学分级、FIGO分期及淋巴结转移密切相关（P0.01）。（2）Kaplan-Meier生存分析显示，宫颈鳞癌组织中NDRG1表达阳性者生存期较长，COX-2及VEGF表达阳性者生存期较短。结论(1)宫颈鳞癌组织中NDRG1表达降低，而COX-2及VEGF表达增加，联合检测宫颈鳞癌组织中NDRG1、COX-2及VEGF的表达可能对判断预后具有重要价值。（2）上调SiHa细胞NDRG1表达后，细胞凋亡增加而细胞增殖降低，其作用机制可能与COX-2、VEGF表达下降有关。
[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.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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