本文選題：CD151蛋白 + 直腸癌��； 參考：《河北醫(yī)科大學》2015年碩士論文

【摘要】：目的:Exosome小體是由多種細胞分泌的攜帶有細胞遺傳物質及蛋白質的小囊泡,在細胞間發(fā)揮物質轉移及信號傳導作用,是近些年人們研究的熱點。Exosome小體可能攜帶某些對免疫功能調(diào)控的信息,可促進腫瘤血管的新生和腫瘤轉移,以及可以直接作用于腫瘤細胞等途徑,影響腫瘤的進展。本課題通過以自身正常粘膜組織做為對照,采用RT㧟PCR法以及Western blot方法檢測直腸癌患者癌組織以及正常粘膜組織中CD151m RNA以及CD151蛋白的表達情況,同步采用Western blot方法檢測直腸癌患者以及健康志愿者尿液exosome小體中CD151蛋白表達情況,并結合臨床病理資料,對比觀察兩種方法檢測CD151基因在直腸癌患者體內(nèi)表達情況以及其與患者臨床病理特征的關系,同時尋求臨床無創(chuàng)檢測該基因表達情況的有效方法。方法:1本研究分別采用RT㧟PCR法以及Western blot檢測法檢測40例直腸癌患者癌組織和癌旁粘膜組織(距癌組織邊緣≥10cm)中CD151m RNA以及CD151蛋白的表達情況。2采用超速離心法進行分離提取直腸癌患者及健康志愿者尿液中的exosome小體,采用Western blot檢測法檢測上述40例直腸癌患者以及40例健康志愿者尿液exosome小體中CD151蛋白表達情況。3最后通過對比患者癌組織與尿液exosome小體中CD151蛋白表達水平的差異,并結合臨床病理資料,分析其與直腸癌患者臨床病理學特征的關系,同時對比兩種檢測方法的優(yōu)劣及可靠性。結果:1直腸癌患者癌組織中CD151m RNA以及CD151蛋白的相對表達量顯著高于癌旁對照組。應用RT㧟PCR法檢測結果顯示,在癌旁對照組中CD151m RNA的相對表達量為0.099±0.012,在直腸癌組中CD151m RNA的相對表達量為0.541±0.056。經(jīng)獨立樣本t檢驗分析,CD151m RNA在直腸癌組織中相對表達量明顯高于癌旁正常組織(P0.05)。應用Western blot檢測法檢測結果顯示,在正常對照組中CD151蛋白相對表達量為0.117±0.007,在直腸癌組中CD151蛋白相對表達量為0.537±0.025。經(jīng)獨立樣本t檢驗分析,CD151蛋白在直腸癌組織中相對表達量明顯高于癌旁正常組織(P0.05)。直腸癌組織中CD151蛋白以及CD151m RNA的相對表達量與患者病理特征進行相關性分析,應用獨立樣本t檢驗分析對數(shù)據(jù)進行處理,結果顯示,CD151m RNA相對表達量以及CD151蛋白的相對表達量與患者直腸癌臨床分期、浸潤深度、分化程度、淋巴結轉移有關,差異有統(tǒng)計學意義(P0.05),與患者年齡、性別無關,差異無統(tǒng)計學意義(P0.05)。2直腸癌患者尿液exosome小體中CD151蛋白水平的相對表達量顯著高于正常對照組。應用Western blot檢測法檢測結果顯示,在正常對照組中CD151蛋白相對表達量為0.153±0.015,在直腸癌組中CD151蛋白相對表達量為0.366±0.016。經(jīng)獨立樣本t檢驗分析,CD151蛋白在直腸癌患者尿液中相對表達量明顯高于正常對照組(P0.05)。直腸癌患者尿液中CD151蛋白相對表達量與患者病理特征進行相關性分析,應用獨立樣本t檢驗分析對數(shù)據(jù)進行處理。結果顯示,CD151蛋白的相對表達量與患者直腸癌臨床分期、浸潤深度、分化程度、淋巴結轉移有關,差異有統(tǒng)計學意義(P0.05),與患者年齡、性別無關,差異無統(tǒng)計學意義(P0.05)。3對直腸癌組患者癌組織以及尿液exosome小體中CD151蛋白表達率進行卡方檢驗,在40例直腸癌患者癌組織中的陽性率為87.5%,在尿液exosome小體中的陽性率為82.5%,兩組比較?2=0.392,P=0.531,差異沒有統(tǒng)計學意義(P0.05),結果顯示CD151蛋白在直腸癌組織中與尿液exosome小體中的表達陽性率無差異,因此,采用Western blot檢測法檢測尿液exosome小體與癌組織中CD151蛋白表達的情況具有同樣的臨床指導意義。結論:1 CD151可能參與直腸癌的發(fā)生及進展過程,其表達水平與直腸癌病程進展具有密切的關系。直腸癌組織中CD151m RNA的相對表達量以及CD151蛋白的相對表達量與直腸癌臨床分期、浸潤深度、分化程度、淋巴結轉移有關,并且伴隨臨床分期的進展、浸潤深度的增加、淋巴結的轉移,CD151的相對表達量明顯增加。2采用Western blot法檢測尿液exosome小體中CD151蛋白水平的表達情況,與新鮮腫瘤組織檢測結果相似,同樣可反映CD151蛋白水平的表達差異與臨床分期、浸潤深度、分化程度、淋巴結轉移等病理特征的相互關系。3檢測尿液exosome小體中目的基因表達情況的方法,取材方便,結果可靠,并可多次檢測,該方法的不斷完善和推廣,可能為臨床惡性腫瘤的治療和病情監(jiān)測提供有力的指導依據(jù)。
[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.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.37

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