CD151在直肠癌组织及尿液exosome小体中表达的对比研究

发布时间：2018-05-29 22:30

本文选题：CD151蛋白 + 直肠癌　；参考：《河北医科大学》2015年硕士论文

【摘要】：目的:Exosome小体是由多种细胞分泌的携带有细胞遗传物质及蛋白质的小囊泡,在细胞间发挥物质转移及信号传导作用,是近些年人们研究的热点。Exosome小体可能携带某些对免疫功能调控的信息,可促进肿瘤血管的新生和肿瘤转移,以及可以直接作用于肿瘤细胞等途径,影响肿瘤的进展。本课题通过以自身正常粘膜组织做为对照,采用RT㧟PCR法以及Western blot方法检测直肠癌患者癌组织以及正常粘膜组织中CD151m RNA以及CD151蛋白的表达情况,同步采用Western blot方法检测直肠癌患者以及健康志愿者尿液exosome小体中CD151蛋白表达情况,并结合临床病理资料,对比观察两种方法检测CD151基因在直肠癌患者体内表达情况以及其与患者临床病理特征的关系,同时寻求临床无创检测该基因表达情况的有效方法。方法:1本研究分别采用RT㧟PCR法以及Western blot检测法检测40例直肠癌患者癌组织和癌旁粘膜组织(距癌组织边缘≥10cm)中CD151m RNA以及CD151蛋白的表达情况。2采用超速离心法进行分离提取直肠癌患者及健康志愿者尿液中的exosome小体,采用Western blot检测法检测上述40例直肠癌患者以及40例健康志愿者尿液exosome小体中CD151蛋白表达情况。3最后通过对比患者癌组织与尿液exosome小体中CD151蛋白表达水平的差异,并结合临床病理资料,分析其与直肠癌患者临床病理学特征的关系,同时对比两种检测方法的优劣及可靠性。结果:1直肠癌患者癌组织中CD151m RNA以及CD151蛋白的相对表达量显著高于癌旁对照组。应用RT㧟PCR法检测结果显示,在癌旁对照组中CD151m RNA的相对表达量为0.099±0.012,在直肠癌组中CD151m RNA的相对表达量为0.541±0.056。经独立样本t检验分析,CD151m RNA在直肠癌组织中相对表达量明显高于癌旁正常组织(P0.05)。应用Western blot检测法检测结果显示,在正常对照组中CD151蛋白相对表达量为0.117±0.007,在直肠癌组中CD151蛋白相对表达量为0.537±0.025。经独立样本t检验分析,CD151蛋白在直肠癌组织中相对表达量明显高于癌旁正常组织(P0.05)。直肠癌组织中CD151蛋白以及CD151m RNA的相对表达量与患者病理特征进行相关性分析,应用独立样本t检验分析对数据进行处理,结果显示,CD151m RNA相对表达量以及CD151蛋白的相对表达量与患者直肠癌临床分期、浸润深度、分化程度、淋巴结转移有关,差异有统计学意义(P0.05),与患者年龄、性别无关,差异无统计学意义(P0.05)。2直肠癌患者尿液exosome小体中CD151蛋白水平的相对表达量显著高于正常对照组。应用Western blot检测法检测结果显示,在正常对照组中CD151蛋白相对表达量为0.153±0.015,在直肠癌组中CD151蛋白相对表达量为0.366±0.016。经独立样本t检验分析,CD151蛋白在直肠癌患者尿液中相对表达量明显高于正常对照组(P0.05)。直肠癌患者尿液中CD151蛋白相对表达量与患者病理特征进行相关性分析,应用独立样本t检验分析对数据进行处理。结果显示,CD151蛋白的相对表达量与患者直肠癌临床分期、浸润深度、分化程度、淋巴结转移有关,差异有统计学意义(P0.05),与患者年龄、性别无关,差异无统计学意义(P0.05)。3对直肠癌组患者癌组织以及尿液exosome小体中CD151蛋白表达率进行卡方检验,在40例直肠癌患者癌组织中的阳性率为87.5%,在尿液exosome小体中的阳性率为82.5%,两组比较?2=0.392,P=0.531,差异没有统计学意义(P0.05),结果显示CD151蛋白在直肠癌组织中与尿液exosome小体中的表达阳性率无差异,因此,采用Western blot检测法检测尿液exosome小体与癌组织中CD151蛋白表达的情况具有同样的临床指导意义。结论:1 CD151可能参与直肠癌的发生及进展过程,其表达水平与直肠癌病程进展具有密切的关系。直肠癌组织中CD151m RNA的相对表达量以及CD151蛋白的相对表达量与直肠癌临床分期、浸润深度、分化程度、淋巴结转移有关,并且伴随临床分期的进展、浸润深度的增加、淋巴结的转移,CD151的相对表达量明显增加。2采用Western blot法检测尿液exosome小体中CD151蛋白水平的表达情况,与新鲜肿瘤组织检测结果相似,同样可反映CD151蛋白水平的表达差异与临床分期、浸润深度、分化程度、淋巴结转移等病理特征的相互关系。3检测尿液exosome小体中目的基因表达情况的方法,取材方便,结果可靠,并可多次检测,该方法的不断完善和推广,可能为临床恶性肿瘤的治疗和病情监测提供有力的指导依据。
[Abstract]:Objective: Exosome corpuscle is a small vesicle which is secreted by a variety of cells and carries a cell genetic material and protein. It plays the role of material transfer and signal transduction in the cells. It is a hot spot in recent years to carry out some information on the regulation of immune function, which can promote the neovascularization and metastasis of tumor vessels, as well as the metastasis of tumor vessels. RT? PCR and Western blot methods were used to detect the expression of CD151m RNA and CD151 protein in the cancer tissues of rectal cancer patients and the normal mucosa tissues by using the normal mucosal tissue as the control, and the CD151m RNA and the CD151 protein in the normal mucosa tissues were detected by the method of Western blot. The expression of CD151 protein in the urine exosome body of the cancer patients and the healthy volunteers, combined with the clinicopathological data, compared the two methods to detect the expression of the CD151 gene in the patients with rectal cancer and the relationship with the clinicopathological features of the patients, and to find an effective method for the noninvasive detection of the gene expression in the patients. Methods: 1 RT PCR and Western blot were used to detect the expression of CD151m RNA and CD151 protein in the cancerous tissues and adjacent mucosa tissues of the rectal cancer patients (the margins of the cancer tissue more than 10cm). The exosome bodies in the urine of the rectal cancer patients and the healthy volunteers were separated and extracted by speeding centrifugation. Western blot detection was used to detect the expression of CD151 protein in the urine exosome corpuscles in 40 cases of rectal cancer and 40 healthy volunteers.3. Finally, the difference in the expression of CD151 protein in the cancer tissues and urine exosome bodies was compared and the clinicopathological features of the patients with rectal cancer were analyzed. The relative expression of CD151m RNA and CD151 protein in the cancer tissues of 1 rectal cancer patients was significantly higher than that in the paracancerous control group. The relative expression of CD151m RNA in the paracancerous control group was 0.099 + 0.012 in the paracancerous control group, and the CD151m RNA phase in the rectal cancer group. The relative expression of CD151m RNA in rectal cancer tissues was significantly higher than that of normal tissue adjacent to cancer (P0.05). The relative expression of CD151 protein in the normal control group was 0.117 + 0.007 in the normal control group, and the relative expression of CD151 protein in the rectal cancer group was 0. in the rectal cancer group. The relative expression of CD151m RNA in rectal cancer was 0.541 + 0.056. test. The relative expression of CD151 protein in rectal cancer tissues was significantly higher than that of normal tissue adjacent to cancer (P0.05) by independent sample t test. The relative expression of CD151 protein and CD151m RNA in rectal cancer tissues was correlated with the pathological features of the patients, and the data were processed with independent sample t test analysis, and the results showed CD. The relative expression of 151m RNA and the relative expression of CD151 protein were related to the clinical stage of rectal cancer, the depth of invasion, the degree of differentiation and lymph node metastasis, and the difference was statistically significant (P0.05), not related to the age and sex of the patients, and the difference was not statistically significant (P0.05) the relative table of the level of CD151 protein in the urine exosome corpuscles of the rectal cancer patients (P0.05). The results of Western blot detection showed that the relative expression of CD151 protein in the normal control group was 0.153 + 0.015, and the relative expression of CD151 protein in the rectal cancer group was 0.366 + 0.016. by independent sample t test, and the relative expression of CD151 protein in the urine of patients with colorectal cancer was significantly higher than that of the normal control group. Normal control group (P0.05). The relative expression of CD151 protein in the urine of rectal cancer patients was correlated with the pathological features of the patients. The data were processed by independent sample t test analysis. The results showed that the relative expression of CD151 protein was related to the clinical stage of rectal cancer, the depth of dipping, the degree of differentiation, and lymph node metastasis. Statistical significance (P0.05) was not related to age and sex of the patients. There was no statistical significance (P0.05). The CD151 expression rate of CD151 protein in the cancer tissues and urine exosome bodies of the rectal cancer patients was examined. The positive rate in the cancer tissues of 40 patients with rectal cancer was 87.5%, the positive rate in the urine exosome body was 82.5%, and the two group was compared to 2=. 0.392, P=0.531, the difference was not statistically significant (P0.05), the results showed that the expression of CD151 protein in rectal cancer tissues and urine exosome corpuscle positive rate was not different, therefore, the use of Western blot detection method for detecting the urine exosome corpuscle and the expression of CD151 protein in the cancer tissue is of the same clinical guiding significance. Conclusion: 1 CD151 can be found. The expression level of the rectal cancer is closely related to the progression of rectal cancer. The relative expression of CD151m RNA and the relative expression of CD151 protein in rectal cancer are related to the clinical stage of rectal cancer, depth of invasion, degree of differentiation, lymph node metastasis, and the progression of clinical stages. The increase of the depth, the metastasis of lymph nodes, the relative expression of CD151 increased obviously by.2, and the expression of CD151 protein in the exosome corpuscle of urine was detected by Western blot, which was similar to the results of fresh tumor tissue detection. It also reflected the difference of expression of CD151 protein and the stage of bed, the depth of infiltration, the degree of differentiation, and the metastasis of lymph nodes. The method of detecting the expression of the target gene in the urine exosome corpuscle, such as the correlation of pathological features, is convenient, the result is reliable, and can be detected many times. The continuous improvement and popularization of this method may provide a powerful guide for the treatment and monitoring of the clinical malignant tumor.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.37

【参考文献】