关于膀胱癌新型标志物MMP1的研究

发布时间：2018-05-10 17:20

本文选题：关于 + 膀胱癌　；参考：《吉林大学》2015年博士论文

【摘要】：在膀胱癌患者尿液中的MMP1（基质金属蛋白酶1）最近被证明同膀胱癌进程密切相关，可以做为新型膀胱癌生物标志物。MMPs的小分子探针检测是一种非侵入性的，高灵敏度和特异性，，且操作简便，费用低廉的检测技术，但是尿液中的MMP1检测还不具备小分子探针，目前只能通过传统的、费时的方法，如酶联免疫吸附分析和免疫印迹。明胶酶谱方法是一种替代小分子探针检测的高敏性MMP1检测方法。本文通过分离人胚胎肾细胞（HEK293）中MMP1基因的cDNA序列，在大肠杆菌中表达了分子探针的筛选靶点MMP1，并对体外表达MMP1和尿液中（人体内源表达）MMP1进行了酶谱分析。研究发现融合TrxA蛋白有助于获得高纯度的可溶型MMP1；对可溶的MMP1和包涵体复性的MMP1以及尿液中的MMP1活性进行明胶酶谱分析，结果显示可溶MMP1比复性MMP1对明胶的降解活性增加了1.54倍，并且更接近尿液中的MMP1。因此，本研究利用明胶酶谱成功检测了体外和体内MMP1的活性，并且为筛选MMP1的小分子探针提供了一个可靠的分子靶点。
[Abstract]:MMP1 (matrix metalloproteinase 1) in urine of patients with bladder cancer has recently been proved to be closely related to the process of bladder cancer. A small molecular probe for.MMPs, a novel biomarker for bladder cancer, is a noninvasive, highly sensitive and specific, easy to operate and inexpensive detection technique, but MMP1 detection in urine There are no small molecular probes, but only by traditional and time-consuming methods, such as enzyme linked immunosorbent assay and immunoblotting. The gelatinase method is a Gao Min MMP1 detection method instead of small molecular probe detection. In this paper, the cDNA sequence of MMP1 gene in human embryonic kidney cells (HEK293) was isolated and expressed in Escherichia coli. The screening target of the subprobe MMP1, and the analysis of the enzyme spectrum in MMP1 and urine (human endogenous expression) MMP1 in vitro. The study found that the fusion of TrxA protein helps to obtain the high purity of soluble MMP1; the soluble MMP1 and inclusion body refolding MMP1 and the activity of MMP1 in the urine are analyzed by gelatinase spectrum, and the results show the soluble MMP1 ratio. The degrading activity of MMP1 to gelatin increased by 1.54 times and was closer to the MMP1. in urine. Therefore, this study successfully detected the activity of MMP1 in vitro and in vivo using gelatinase spectrum, and provided a reliable molecular target for the screening of small molecular probes for MMP1.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.14

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