Wnt3a及Lgr5在膀胱癌中的表达及意义

发布时间：2018-12-28 12:53

【摘要】：目的：膀胱癌是所有泌尿系肿瘤中最常见的，，其发病率正在逐年上升，对人类的生命健康构成严重威胁。膀胱癌的发生发展是极其复杂的一个过程，尽管进行了大量的研究，但其发生机制目前还不明确。 Wnt信号通路是细胞中高度保守的信号传导通路，参与调控细胞的增殖、分化和凋亡等生物过程，是有多种分子共同参与的多个步骤、多个作用位点的复杂的信号转导通路。近几年研究发现，Wnt通路的过度激活与胃癌、肝癌、结肠癌等多种恶性肿瘤的发生发展密切相关。作为激活Wnt信号通路启动因子的Wnt蛋白家族成员已经发现19种，他们属于富含半胱氨酸的分泌型糖蛋白。Wnt3a属于启动因子中的一员，能够稳定的激活经典Wnt信号通路。Lgr5又名GPR49是新近发现的Wnt信号通路的靶基因，属于G蛋白偶联受体家族的成员。Lgr5在多种组织干细胞中表达，其作为干细胞标记物已被证实，并且有证据表明其作为Wnt信号通路的下游靶基因参与了肿瘤的发生发展。干细胞标记物的研究是目前肿瘤研究的焦点，因此本文初步研究Wnt3a、Lgr5在膀胱癌组织中的表达，探讨Wnt3a、Lgr5在人膀胱癌组织中的表达和意义，旨在为膀胱癌的治疗提供新的靶点。 方法：搜集2012年4月-2013年10月在河北医科大学第二医院手术治疗患者中的53例膀胱癌组织和13例正常膀胱组织，依据国际抗癌联盟2009年TNM分期标准将膀胱癌组织分为两组：非肌层浸润性膀胱癌34例，肌层浸润性膀胱癌19例。同时取膀胱癌组织17例，正常膀胱组织9例于术后立即保存于-80℃中备用。应用免疫组织化学法检测13例正常膀胱组织和53例膀胱癌组织Wnt3a、 Lgr5蛋白表达情况，并进一步观察在非肌层浸润性膀胱癌及肌层浸润性膀胱癌中表达的差异性。RT-PCR方法检测9例正常膀胱组织和17例膀胱癌组织中Wnt3a、 Lgr5mRNA的表达情况。应用SPSS13.0统计软件进行统计，采用卡方检验Fisher’s精确概率法分别比较Wnt3a和Lgr5两组间阳性表达率差异，每个样本Wnt3a与Lgr5的计分采用pearson相关分析法分析两者的关系，吸光度比值采用t检验进行统计分析 结果： 1免疫组化结果显示：Wnt3a蛋白在膀胱癌与正常膀胱组织表达阳性率具有显著差异性，分别为71.7%及30.8%；在非肌层浸润性和肌层浸润性膀胱癌中的表达阳性率分别为61.8%、89.5%，差异均有统计学意义（P＜0.05）；Lgr5蛋白在膀胱癌与正常膀胱组织表达阳性率分别为64.2%及23.1%，在非肌层浸润性和肌层浸润性膀胱癌中表达阳性率分别为52.9%、84.2%，差异有均统计学意义（P＜0.05）；两者在膀胱癌中的表达呈正相关（r=0.659，P＜0.01）。 2RT-PCR结果显示：Wnt3a在膀胱癌和正常膀胱组织的光密度比值分别为0.197±0.029和0.088±0.019，差异有统计学意义（P＜0.05)；Lgr5在膀胱癌和正常膀胱组织的光密度比值分别为0.408±0.061和0.161±0.053，差异有统计学意义（P＜0.05）。 结论：在膀胱癌组织中Wnt3a、 Lgr5蛋白及mRNA的表达均高于正常膀胱组织，提示Wnt3a、 Lgr5可能与膀胱癌的发生发展密切相关。Wnt3a、 Lgr5蛋白在肌层浸润性膀胱癌中的表达阳性率明显高于其在非肌层浸润性膀胱癌中的表达阳性率，表明Wnt3a、 Lgr5蛋白可能与膀胱癌的侵袭性有关。
[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

本文编号：2393966