發(fā)布時(shí)間：2018-05-10 17:20

  本文選題：關(guān)于 + 膀胱癌��； 參考：《吉林大學(xué)》2015年博士論文

【摘要】：在膀胱癌患者尿液中的MMP1（基質(zhì)金屬蛋白酶1）最近被證明同膀胱癌進(jìn)程密切相關(guān)，可以做為新型膀胱癌生物標(biāo)志物。MMPs的小分子探針檢測(cè)是一種非侵入性的，高靈敏度和特異性，，且操作簡(jiǎn)便，費(fèi)用低廉的檢測(cè)技術(shù)，但是尿液中的MMP1檢測(cè)還不具備小分子探針，目前只能通過(guò)傳統(tǒng)的、費(fèi)時(shí)的方法，如酶聯(lián)免疫吸附分析和免疫印跡。明膠酶譜方法是一種替代小分子探針檢測(cè)的高敏性MMP1檢測(cè)方法。本文通過(guò)分離人胚胎腎細(xì)胞（HEK293）中MMP1基因的cDNA序列，在大腸桿菌中表達(dá)了分子探針的篩選靶點(diǎn)MMP1，并對(duì)體外表達(dá)MMP1和尿液中（人體內(nèi)源表達(dá)）MMP1進(jìn)行了酶譜分析。研究發(fā)現(xiàn)融合TrxA蛋白有助于獲得高純度的可溶型MMP1；對(duì)可溶的MMP1和包涵體復(fù)性的MMP1以及尿液中的MMP1活性進(jìn)行明膠酶譜分析，結(jié)果顯示可溶MMP1比復(fù)性MMP1對(duì)明膠的降解活性增加了1.54倍，并且更接近尿液中的MMP1。因此，本研究利用明膠酶譜成功檢測(cè)了體外和體內(nèi)MMP1的活性，并且為篩選MMP1的小分子探針提供了一個(gè)可靠的分子靶點(diǎn)。
[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.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R737.14

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