ATP5A1在大肠癌中的表达及其临床意义

发布时间：2019-05-23 20:15

【摘要】：研究背景及目的:大肠癌(Colorectal cancer,CRC),别名为结直肠癌,是一种全球范围内发病率和死亡率都很高的疾病,是世界第五大最常见的肿瘤,第三大全球肿瘤相关死因[1],其五年生存率小于65%,一般患者确诊时已处于晚期阶段,从而给治疗成功率带来了很大的限制。有研究显示,25%的患者确诊时已经发生了转移[2]。疾病的早期诊断,对患者生活质量提高有很大价值。所以亟需一个灵敏度和特异度都高的肿瘤标志物,来实现对结直肠癌的早发现、早诊断、早治疗。利用二维色谱与质谱联用技术的蛋白组学,可以发现癌症组织与相应癌旁组织中蛋白质表达的差异,是我们探求新的标志物强有力的技术支持。本研究通过二维色谱与质谱联用技术,分析了大肠癌与癌旁组织之间蛋白表达的不同,构建大肠癌的差异蛋白质表达谱,期望从中选择出对大肠癌的诊断及治疗有很高价值的标志物。方法:本研究主要分下面几个部分进行:1.二维色谱与质谱联用技术建立大肠癌及相应癌旁组织差异蛋白谱。方法:1)选取大肠癌腺癌的癌组织及其相应癌旁组织标本,提取组织蛋白,制备多肽混合物。2)应用二维色谱及质谱联用技术检测癌组织及相应癌旁组织差异表达蛋白。3)分析得到大肠癌及相应癌旁组织差异表达蛋白谱。2.验证ATP5A1在大肠癌中的表达。方法:选取3组大肠癌腺癌的癌组织及其相应癌旁组织标本,用Western Blot的方法印证目标蛋白ATP5A1在大肠癌及癌旁组织的表达。3.大肠癌患者ATP5A1表达情况与临床资料之间的关系。方法:1)从患者的年龄、性别、TNM分期及有无远处转移等方面建立临床病理资料数据库。2)应用免疫组化方法检测ATP5A1在大肠癌中的表达情况,同时分析其表达情况与患者的临床资料之间的关系。结果:1.建立大肠癌及相应癌旁组织差异蛋白谱,分析得出差异表达的蛋白质25个,其中18个蛋白在癌组织中表达上调,7个蛋白在癌组织中表达下调。2.Western Blot结果显示大肠癌组织中ATP5A1的表达量较癌旁组织明显升高,进一步验证了第一部分蛋白组学的结果。3.⑴ATP5A1在大肠癌癌组织中阳性表达,主要表达在胞浆和胞膜,而在相应癌旁组织中呈阴性表达。⑵20例大肠癌组织中,7例ATP5A1蛋白表达阳性,得出阳性率为35%;而20例相应癌旁组织中,未见阳性表达。这表明,大肠癌组织中ATP5A1蛋白的表达水平比相应的癌旁组织明显上调,差异有统计学意义(p0.05)。⑶ATP5A1蛋白的表达与大肠癌组织TNM分期、远处转移有关,并且与患者血清中CEA表达水平高度相关(p0.05)。但是与患者的年龄、性别、临床诊断及分化程度均无关(p0.05)。结论:1.大肠癌组织中ATP5A1的表达量较癌旁组织明显升高。2.ATP5A1蛋白的表达与大肠癌组织TNM分期、远处转移有关,并且与患者血清中CEA表达水平高度相关。但是与患者的年龄、性别、临床诊断及分化程度均无关。3.ATP5A1在大肠癌肿瘤发生及进展中发挥重要作用,对大肠癌恶性程度诊断有一定价值,为进一步研究奠定了基础。
[Abstract]:Background and purpose of the study: Colorectal cancer (CRC), an alias of colorectal cancer, is a disease with high morbidity and mortality in the world. It is the fifth most common tumor in the world, the third largest global tumor-related cause of death[1], and the five-year survival rate is less than 65%. In general, the patient is in the advanced stage at the time of diagnosis, so that the success rate of the treatment is greatly limited. A study showed that 25% of patients had been transferred[2] at the time of diagnosis. The early diagnosis of the disease is of great value to the improvement of the quality of the patient's life. Therefore, a tumor marker with high sensitivity and specificity is needed to realize early detection, early diagnosis and early treatment of colorectal cancer. By using the proteomics of two-dimensional chromatography and mass spectrometry, it can be found that the difference between the expression of the proteins in the cancer tissues and the corresponding adjacent tissues is a powerful technical support for us to search for new markers. In this study, by means of two-dimensional chromatography and mass spectrometry, the expression of the protein between the colorectal cancer and the adjacent tissue is analyzed, and the differential protein expression profile of the colorectal cancer is constructed, and it is expected to select a high-value marker for the diagnosis and treatment of the large intestine cancer. Methods: The main part of this study was as follows:1. The differential protein spectrum of colorectal cancer and corresponding adjacent tissues was established by two-dimensional chromatography and mass spectrometry. method:1) selecting a cancer tissue of a large intestine cancer and a corresponding adjacent tissue sample to extract the tissue protein, 2) The expression of the differentially expressed proteins in the cancer tissues and the corresponding adjacent tissues was detected by two-dimensional chromatography and mass spectrometry. To verify the expression of CYP5A1 in colorectal cancer. Methods: The expression of target protein A5A1 in colorectal cancer and adjacent tissues was confirmed by Western Blot method. The relationship between the expression of CYP5A1 and clinical data in patients with colorectal cancer. Methods:1) The clinical and pathological data were established from the age, sex, TNM stage of the patient and the presence or absence of distant metastasis. Results:1. 2. Western Blot showed that the expression of CYP5A1 in the tissues of colorectal cancer was significantly higher than that of the adjacent tissues. The results of the first partial proteomics are further validated. The positive expression of Astragalus 5A1 in the carcinoma of the large intestine is mainly expressed in the cytoplasm and the cell membrane, while in the corresponding adjacent tissues, the expression of the negative expression is negative. In 20 cases of colorectal cancer,7 cases of H5A1 protein were positive, and the positive rate was 35%; in 20 of the 20 corresponding adjacent tissues, no positive expression was found. The expression of CYP5A1 in colorectal cancer was significantly higher than that of the corresponding adjacent tissues, and the difference was statistically significant (p0.05). The expression of the CYP5A1 protein was related to the TNM stage and distant metastasis of the colorectal cancer tissue, and was highly correlated with the level of CEA expression in the serum of the patient (p0.05). However, it was not related to the age, sex, clinical diagnosis and differentiation of the patient (p0.05). Conclusion:1. The expression of CYP5A1 in colorectal cancer was significantly higher than that of the adjacent tissues. The expression of the A5A1 protein was related to the TNM stage and distant metastasis of the colorectal cancer tissues, and was highly correlated with the level of CEA expression in the serum of the patient. 3. A5A1 plays an important role in the tumorigenesis and progression of the large intestine, and has a certain value for the diagnosis of the degree of malignancy of the large intestine, and lays the foundation for further study.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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