本文選題：乳腺癌 + STIM1��； 參考：《天津醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的乳腺癌是女性最常見(jiàn)的惡性腫瘤之一,近年來(lái)發(fā)病率在環(huán)球內(nèi)呈上升的趨向,其發(fā)生和轉(zhuǎn)移是一個(gè)與多因素相關(guān)的復(fù)雜過(guò)程。經(jīng)研究發(fā)現(xiàn),STIM1蛋白是構(gòu)成鈣庫(kù)操縱性鈣通道(store-opetated Ca2+channels,SOCs)的主要成分之一,是細(xì)胞內(nèi)鈣離子的傳感器,可以使細(xì)胞外的鈣離子進(jìn)入細(xì)胞內(nèi),從而參與乳腺癌的侵襲和轉(zhuǎn)移過(guò)程。micro RNA是一類由內(nèi)源基因編碼的非編碼單鏈RNA分子,參與生命活動(dòng)中的重要進(jìn)程,約調(diào)控人類三分之一的基因。有些miRNAs可以通過(guò)轉(zhuǎn)錄后負(fù)性調(diào)控致癌性靶基因表達(dá)的方式而發(fā)揮抑癌功能;相反,有一部分miRNAs可以通過(guò)下調(diào)抑癌靶基因表達(dá)的方式起到癌基因的作用。在前期工作中,我們小組利用免疫組化的方法發(fā)現(xiàn)STIM1蛋白在乳腺IDC組織中呈高表達(dá)狀態(tài),且STIM1高表達(dá)是乳腺癌患者預(yù)后的一個(gè)不良指標(biāo)。經(jīng)生物信息學(xué)預(yù)測(cè)miR-223-3p和miR-150-5p有可能直接調(diào)控STIM1基因,從而在乳腺癌的疾病進(jìn)程中發(fā)揮作用。目前在乳腺IDC中miR-223-3p和miR-150-5p與STIM1基因的關(guān)系及作用機(jī)制尚不清楚,因此本課題旨在研究miR-223-3p-STIM1及miR-150-5p-STIM1在乳腺癌中的相互作用以及兩個(gè)miRNA和STIM1蛋白與乳腺癌臨床病理資料和預(yù)后的關(guān)系,為乳腺癌病人提供新的預(yù)后標(biāo)志物,并且為乳腺癌的綜合治療需找新的靶點(diǎn)。方法本題選取天津市腫瘤醫(yī)院2007年1月至2009年12月期間的原發(fā)性乳腺IDC患者110例以及其中癌和對(duì)應(yīng)癌旁組織的標(biāo)本60例,通過(guò)查閱病歷的方式獲取患者的臨床病理資料,通過(guò)電話隨訪或者門診電子檢查結(jié)果獲取隨訪信息。通過(guò)構(gòu)建雙熒光素酶報(bào)告基因的方法驗(yàn)證STIM1是miR-223-3p和miR-150-5p直接調(diào)控的靶基因,此實(shí)驗(yàn)重復(fù)三次。采用乳腺癌組織切片原位雜交的方法檢測(cè)miR-223-3p和miR-150-5p在乳腺癌組織及相應(yīng)的癌旁組織中的表達(dá)量,用Kaplan-Meier法分析miR-223-3p、miR-150-5p和STIM1蛋白與乳腺癌患者總生存時(shí)間和無(wú)病生存時(shí)間的相關(guān)性,用卡方檢驗(yàn)分析miR-223-3p、miR-150-5p和STIM1蛋白與患者臨床病理資料的關(guān)系。根據(jù)前期中我們用免疫組化方法測(cè)定的乳腺癌組織中的STIM1蛋白的表達(dá)結(jié)果,用Spearman相關(guān)法分析miR-223-3p和miR-150-5p分別與STIM1蛋白的相關(guān)性,P0.05具有統(tǒng)計(jì)學(xué)差異。結(jié)果1.結(jié)果顯示,在miR-223-3p組中,在MDA-MB-231和MCF-7兩個(gè)乳腺癌細(xì)胞系中轉(zhuǎn)染STIM1-3,UTR-WT后熒光素酶的相對(duì)熒光值明顯下降,而轉(zhuǎn)染STIM1-3,UTR-MT后熒光素酶的相對(duì)熒光值沒(méi)有明顯變化(P0.05)。說(shuō)明miR-223-3p可以直接與STIM1-3,UTR區(qū)結(jié)合,從而影響其生物學(xué)功能。在MDA-MB-231和MCF-7乳腺癌細(xì)胞系中轉(zhuǎn)染STIM1-3‘UTR-WT/miR-150-5p mimic與STIM1-3,UTR-MT/miR-150-5p mimic的相對(duì)熒光值差異并沒(méi)有統(tǒng)計(jì)學(xué)意義,P值分別為0.754和0.458。2.miR-223-3p在乳腺癌組織中表達(dá)較低,高表達(dá)水平的乳腺癌患者的預(yù)后優(yōu)于低表達(dá)水平的患者(OS:P=0.038;DFS:P=0.196)。在乳腺癌組織中miR-223-3p表達(dá)水平與STIM1蛋白的表達(dá)水平呈負(fù)相關(guān)(r=-1.914,P=0.043)。miR-223-3p的表達(dá)水平與患者的淋巴結(jié)轉(zhuǎn)移(P=0.035)呈顯著相關(guān)性。3.miR-150-5p在乳腺癌組織中呈高表達(dá),表達(dá)較低的乳腺癌患者的預(yù)后有優(yōu)于表達(dá)高的患者的趨勢(shì),(OS:P=0.734;DFS:P=0.077)。在乳腺癌組織中miR-150-5p表達(dá)水平與STIM1蛋白的表達(dá)水平不具有顯著的相關(guān)性(r=0.001,P=0.991)。miR-150-5p表達(dá)水平與乳腺腫瘤大小(P=0.035)、TNM病理分期(P=0.003)以及ER狀態(tài)(P=0.035)具有顯著的關(guān)聯(lián)性。結(jié)論1.miR-223-3p在乳腺浸潤(rùn)性導(dǎo)管在組織中表達(dá)水平較低,低表達(dá)的乳腺癌患者更容易出現(xiàn)淋巴結(jié)轉(zhuǎn)移,miR-223-3p高表達(dá)組的患者較低表達(dá)的患者預(yù)后好,STIM1是miR-223-3p直接調(diào)控的靶基因,有可能成為治療乳腺癌的新藥物。2.在乳腺浸潤(rùn)性導(dǎo)管癌組織中,miR-150-5p表達(dá)較高,miR-150-5p低表達(dá)的乳腺癌患者有優(yōu)于高表達(dá)的趨勢(shì)。miR-150-5p表達(dá)水平與患者腫瘤大小和乳腺癌TNM分期以及ER相關(guān),提示miR-150-5p在乳腺癌的侵襲和轉(zhuǎn)移中起到重要作用。
[Abstract]:Objective breast cancer is one of the most common malignant tumors in women. In recent years the incidence of the disease is increasing in the world. Its occurrence and metastasis is a complex process related to multiple factors. It is found that STIM1 protein is one of the main components of store-opetated Ca2+ channels (SOCs), which is the intracellular calcium ionization. The subcellular sensor, which can make the extracellular calcium ions into the cell, participates in the invasion and metastasis of breast cancer, and.Micro RNA is a class of non coded single strand RNA molecules encoded by endogenous genes. It participates in important processes in life activities and regulates human 1/3. Some miRNAs can be induced by post transcriptional negative regulation. On the contrary, a part of miRNAs can play the role of Oncogene by down regulating the expression of tumor suppressor gene. In the early work, our group used immunohistochemical method to find the high expression of STIM1 protein in IDC tissues of the breast, and the high expression of STIM1 is the breast cancer. It is predicted that miR-223-3p and miR-150-5p may directly regulate the STIM1 gene and play a role in the process of breast cancer by bioinformatics. The relationship and mechanism of miR-223-3p and miR-150-5p with the STIM1 gene are still unclear in breast IDC. Therefore, this subject is aimed at studying miR-223-3p-STIM1 The interaction of miR-150-5p-STIM1 in breast cancer and the relationship between two miRNA and STIM1 proteins and the clinicopathological data and prognosis of breast cancer provide a new prognostic marker for breast cancer patients and need to find new targets for the comprehensive treatment of breast cancer. Method this problem was selected from Tianjin Tumour Hospital from January 2007 to December 2009. 110 cases of primary mammary IDC and 60 cases of carcinoma and paracancerous tissue were obtained. The clinicopathological data of the patients were obtained by consulting the medical records. The follow-up information was obtained by telephone follow-up or outpatient electronic examination. The method of constructing the double luciferase reporter gene was used to verify that STIM1 was miR-223-3p and miR-150-5p. The target gene was regulated directly. This experiment was repeated three times. The expression of miR-223-3p and miR-150-5p in breast cancer tissues and corresponding para cancerous tissues was detected by breast cancer tissue section in situ hybridization. The total survival time and disease-free survival time of miR-223-3p, miR-150-5p, STIM1 protein and breast cancer patients were analyzed by Kaplan-Meier method. Correlation, the relationship between miR-223-3p, miR-150-5p and STIM1 protein with the patient's clinicopathological data was analyzed with the chi square test. According to the expression of STIM1 protein in the breast cancer tissue that we used in the early stage of immunohistochemistry, the correlation of miR-223-3p and miR-150-5p with STIM1 protein was analyzed by Spearman correlation method, and P0.05 has a series of data. Results 1. results showed that in group miR-223-3p, STIM1-3 was transfected into two breast cancer cell lines in MDA-MB-231 and MCF-7, and the relative fluorescence value of luciferase was obviously decreased after UTR-WT, while STIM1-3 was transfected with STIM1-3, and the relative fluorescence of luciferase was not significantly changed after UTR-MT (P0.05). It indicated that miR-223-3p could be directly associated with STIM1-3, UTR region. In MDA-MB-231 and MCF-7 breast cancer cell lines transfected with STIM1-3 'UTR-WT/miR-150-5p mimic and STIM1-3, the relative fluorescence values of UTR-MT/miR-150-5p mimic are not statistically significant, and P values are 0.754 and 0.458.2.miR-223-3p in breast cancer tissues that express low, high expression levels of breast cancer. The prognosis of patients was better than that of low expression level (OS:P=0.038; DFS:P=0.196). The expression level of miR-223-3p in breast cancer tissues was negatively correlated with the expression level of STIM1 protein (r=-1.914, P=0.043).MiR-223-3p expression level was significantly correlated with lymph node metastasis (P=0.035) in patients with high expression of.3.miR-150-5p in breast cancer tissue. The prognosis of patients with low expression of breast cancer is better than that of high expression patients (OS:P=0.734; DFS:P=0.077). There is no significant correlation between the expression level of miR-150-5p and the expression level of STIM1 protein in breast cancer tissues (r=0.001, P=0.991).MiR-150-5p expression and breast tumor size (P=0.035), TNM pathological staging (P=0.003) ER state (P=0.035) has a significant correlation. Conclusion the expression level of 1.miR-223-3p in mammary gland infiltrating ducts is low. The patients with low expression of breast cancer are more prone to lymph node metastasis, and the patients with miR-223-3p high expression group have better prognosis. STIM1 is the target gene directly regulated by miR-223-3p, and it may become a cure. .2., a new drug for breast cancer, has a high expression of miR-150-5p in invasive ductal carcinoma of the breast. The expression level of breast cancer with low expression of miR-150-5p is superior to that of high expression of.MiR-150-5p, which is associated with the size of the tumor and the TNM staging of the breast cancer and ER, suggesting that miR-150-5p plays an important role in the invasion and metastasis of breast cancer.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R737.9
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本文編號(hào)：1943004