【摘要】：目的：膀胱癌是所有泌尿系腫瘤中最常見的，，其發(fā)病率正在逐年上升，對人類的生命健康構(gòu)成嚴重威脅。膀胱癌的發(fā)生發(fā)展是極其復(fù)雜的一個過程，盡管進行了大量的研究，但其發(fā)生機制目前還不明確。 Wnt信號通路是細胞中高度保守的信號傳導(dǎo)通路，參與調(diào)控細胞的增殖、分化和凋亡等生物過程，是有多種分子共同參與的多個步驟、多個作用位點的復(fù)雜的信號轉(zhuǎn)導(dǎo)通路。近幾年研究發(fā)現(xiàn)，Wnt通路的過度激活與胃癌、肝癌、結(jié)腸癌等多種惡性腫瘤的發(fā)生發(fā)展密切相關(guān)。作為激活Wnt信號通路啟動因子的Wnt蛋白家族成員已經(jīng)發(fā)現(xiàn)19種，他們屬于富含半胱氨酸的分泌型糖蛋白。Wnt3a屬于啟動因子中的一員，能夠穩(wěn)定的激活經(jīng)典Wnt信號通路。Lgr5又名GPR49是新近發(fā)現(xiàn)的Wnt信號通路的靶基因，屬于G蛋白偶聯(lián)受體家族的成員。Lgr5在多種組織干細胞中表達，其作為干細胞標記物已被證實，并且有證據(jù)表明其作為Wnt信號通路的下游靶基因參與了腫瘤的發(fā)生發(fā)展。干細胞標記物的研究是目前腫瘤研究的焦點，因此本文初步研究Wnt3a、Lgr5在膀胱癌組織中的表達，探討Wnt3a、Lgr5在人膀胱癌組織中的表達和意義，旨在為膀胱癌的治療提供新的靶點。 方法：搜集2012年4月-2013年10月在河北醫(yī)科大學(xué)第二醫(yī)院手術(shù)治療患者中的53例膀胱癌組織和13例正常膀胱組織，依據(jù)國際抗癌聯(lián)盟2009年TNM分期標準將膀胱癌組織分為兩組：非肌層浸潤性膀胱癌34例，肌層浸潤性膀胱癌19例。同時取膀胱癌組織17例，正常膀胱組織9例于術(shù)后立即保存于-80℃中備用。應(yīng)用免疫組織化學(xué)法檢測13例正常膀胱組織和53例膀胱癌組織Wnt3a、 Lgr5蛋白表達情況，并進一步觀察在非肌層浸潤性膀胱癌及肌層浸潤性膀胱癌中表達的差異性。RT-PCR方法檢測9例正常膀胱組織和17例膀胱癌組織中Wnt3a、 Lgr5mRNA的表達情況。應(yīng)用SPSS13.0統(tǒng)計軟件進行統(tǒng)計，采用卡方檢驗Fisher’s精確概率法分別比較Wnt3a和Lgr5兩組間陽性表達率差異，每個樣本W(wǎng)nt3a與Lgr5的計分采用pearson相關(guān)分析法分析兩者的關(guān)系，吸光度比值采用t檢驗進行統(tǒng)計分析 結(jié)果： 1免疫組化結(jié)果顯示：Wnt3a蛋白在膀胱癌與正常膀胱組織表達陽性率具有顯著差異性，分別為71.7%及30.8%；在非肌層浸潤性和肌層浸潤性膀胱癌中的表達陽性率分別為61.8%、89.5%，差異均有統(tǒng)計學(xué)意義（P＜0.05）；Lgr5蛋白在膀胱癌與正常膀胱組織表達陽性率分別為64.2%及23.1%，在非肌層浸潤性和肌層浸潤性膀胱癌中表達陽性率分別為52.9%、84.2%，差異有均統(tǒng)計學(xué)意義（P＜0.05）；兩者在膀胱癌中的表達呈正相關(guān)（r=0.659，P＜0.01）。 2RT-PCR結(jié)果顯示：Wnt3a在膀胱癌和正常膀胱組織的光密度比值分別為0.197±0.029和0.088±0.019，差異有統(tǒng)計學(xué)意義（P＜0.05)；Lgr5在膀胱癌和正常膀胱組織的光密度比值分別為0.408±0.061和0.161±0.053，差異有統(tǒng)計學(xué)意義（P＜0.05）。 結(jié)論：在膀胱癌組織中Wnt3a、 Lgr5蛋白及mRNA的表達均高于正常膀胱組織，提示W(wǎng)nt3a、 Lgr5可能與膀胱癌的發(fā)生發(fā)展密切相關(guān)。Wnt3a、 Lgr5蛋白在肌層浸潤性膀胱癌中的表達陽性率明顯高于其在非肌層浸潤性膀胱癌中的表達陽性率，表明Wnt3a、 Lgr5蛋白可能與膀胱癌的侵襲性有關(guān)。
[Abstract]:Objective: Bladder cancer is the most common in all urinary tract tumors. The incidence of bladder cancer is increasing year by year, which is a serious threat to human health. The development of bladder cancer is an extremely complex process, although a large number of studies have been carried out, the mechanism is not yet clear. The Wnt signaling pathway is a highly conserved signaling pathway in the cell, and is involved in the control of the proliferation, differentiation and apoptosis of the cells. Road. In recent years, it has been found that over-activation of the Wnt pathway is closely related to the development of various malignant tumors, such as gastric cancer, liver cancer, and colon cancer. The Wnt protein family member, which is the activation factor for activating the Wnt signaling pathway, has found 19 species, which belong to cysteine-rich secreted sugar eggs. White. Wnt3a is a member of the start-up factor, enabling a stable activation of the classic Wnt signal Lgr5, also known as GPR49, is a newly discovered target gene of the Wnt signaling pathway, which belongs to the G protein-coupled receptor family. The expression of Lgr5 in a variety of tissue stem cells as a stem cell marker has been demonstrated and there is evidence that it is involved in the occurrence of a tumor as a downstream target gene of the Wnt signaling pathway. The research of stem cell marker is the focus of the present tumor research, so this paper first studies the expression of Wnt3a and Lgr5 in the bladder cancer tissue, and probes into the expression and significance of Wnt3a and Lgr5 in the human bladder cancer tissue, aiming at providing a new target for the treatment of bladder cancer. Methods: 53 cases of bladder cancer and 13 normal bladder tissues were collected from April, 2012 to October, 2013 in the second hospital of Hebei Medical University. 34 cases, myolayer infiltrative bladder cancer 19 cases of bladder cancer, 17 cases of bladder cancer and 9 cases of normal bladder tissue were immediately stored at-80 鈩

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