腫瘤細胞內(nèi)活性物質(zhì)的分析和檢測
發(fā)布時間：2019-06-12 11:54
【摘要】：癌癥,一種嚴重威脅著人類生命健康的疾病,隨著人類生存環(huán)境的惡化,其發(fā)病率逐年升高。癌癥患者的治愈率極低,所以癌癥的治療以及早期的偵測檢查成為近些年來醫(yī)學上的一大難題。越早的發(fā)現(xiàn)腫瘤的存在,癌癥患者被治愈的希望就越高。腫瘤細胞內(nèi)活性物質(zhì)的檢測對于癌癥的預(yù)防和診斷治療等有著重要意義。本文通過納米技術(shù)、DNA生物傳感器技術(shù)以及熒光分析檢測技術(shù)為檢測腫瘤細胞內(nèi)活性物提供了新的檢測手段和思路。本文主要內(nèi)容包括以下三個方面：1、提出了一種新穎簡單的制備硅基介孔材料的方法。通過改變在制備過程的物料比來調(diào)節(jié)介孔材料的孔徑結(jié)構(gòu),采用特定物料來對介孔材料進行有機功能化改性,進而改善介孔材料的宏觀物理化學性質(zhì)。提出了將介孔材料作為一種生物納米容器的設(shè)想并將其應(yīng)用到生物化學反應(yīng)體系當中。2、創(chuàng)造了一種利用生物納米容器釋放熒光素分子檢測ATP的熒光分析檢測方法。將適體的互補鏈結(jié)合到納米容器的表面,通過DNA雜交金納米粒子將熒光素分子封閉在納米容器內(nèi)部。加入ATP,適體與ATP分子特異性結(jié)合,使金納米粒子離開納米容器表面,釋放出熒光素分子。通過檢測熒光信號,來實現(xiàn)對ATP的定性定量分析。對于生物納米容器作為藥物載體在癌癥治療中的應(yīng)用做了初步探索。3、設(shè)計了一種新穎的基于熒光檢測技術(shù)的基礎(chǔ)上通過鏈循環(huán)放大技術(shù)檢測端粒酶的方法。將發(fā)卡DNA修飾在磁性納米微球的表面，引入端粒酶前體與發(fā)卡DNA的部分序列互補配對,端粒酶的引入可以將發(fā)卡DNA莖環(huán)結(jié)構(gòu)打開,然后將攜載大量熒光DNA以及少量連接DNA的生物納米探針加入至體系當,當探針固定在磁性納米微球的表面時,在DNA聚合酶的作用下,連接DNA開始生長并將端粒酶前體頂替下來,參與到下一個循環(huán)當中,對信號的檢測起到了明顯的放大作用。由于磁性納米微球可以攜載多個生物納米探針,同時金納米粒子也可攜載多個熒光DNA分子,所以整個實驗對信號實現(xiàn)了多重放大,對端粒酶的檢測也有著較高的靈敏度。
[Abstract]:Cancer, a serious threat to human life and health, with the deterioration of human living environment, its incidence is increasing year by year. The cure rate of cancer patients is very low, so the treatment of cancer and early detection has become a medical problem in recent years. The sooner tumors are discovered, the more likely cancer patients are to be cured. The detection of tumor intracellular active substances is of great significance for the prevention, diagnosis and treatment of cancer. In this paper, nanotechnology, DNA biosensor technology and fluorescence analysis technology provide a new detection method and ideas for the detection of intracellular active substances in tumor cells. The main contents of this paper are as follows: 1. A novel and simple method for preparing silicon-based mesoporous materials is proposed. The pore size structure of mesoporous materials is adjusted by changing the material ratio in the preparation process, and the mesoporous materials are modified organically by using specific materials to improve the macroscopic physical and chemical properties of mesoporous materials. The idea of mesoporous materials as a biological nano-container was proposed and applied to the biochemical reaction system. 2. A fluorescence analysis method for the detection of ATP by releasing fluorescein molecules from biological nano-containers was created. The complementary chain of the aptamer was combined to the surface of the nano-container, and the fluorescein molecule was enclosed in the nano-container by DNA hybrid gold nanoparticles. The ATP, aptamer was specifically combined with ATP molecule to make the gold nanoparticles leave the surface of the nano-container and release fluorescein molecules. The qualitative and quantitative analysis of ATP is realized by detecting fluorescence signal. The application of biological nano-container as drug carrier in cancer treatment was explored. 3. A novel method based on fluorescence detection technology was designed to detect telomerase by chain cycle amplification technique. The hairpin DNA was modified on the surface of magnetic nanoparticles, and the telomerase precursor was complementary to the partial sequence of hairpin DNA. The structure of hairpin DNA stem ring could be opened by the introduction of telomerase, and then a large number of fluorescent DNA and a small amount of biological nano-probes connected to DNA could be added to the system when the probe was fixed on the surface of magnetic nanoparticles under the action of DNA polymerase. The connected DNA began to grow and replaced the telomerase precursor to participate in the next cycle, which played an obvious role in amplifying the signal detection. Because magnetic nanoparticles can carry multiple biological nanoprobes and gold nanoparticles can also carry multiple fluorescent DNA molecules, the whole experiment realizes multiple amplification of signals and has high sensitivity to telomerase detection.
【學位授予單位】：青島科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R730.4;O657.3
本文編號：2498001
[Abstract]:Cancer, a serious threat to human life and health, with the deterioration of human living environment, its incidence is increasing year by year. The cure rate of cancer patients is very low, so the treatment of cancer and early detection has become a medical problem in recent years. The sooner tumors are discovered, the more likely cancer patients are to be cured. The detection of tumor intracellular active substances is of great significance for the prevention, diagnosis and treatment of cancer. In this paper, nanotechnology, DNA biosensor technology and fluorescence analysis technology provide a new detection method and ideas for the detection of intracellular active substances in tumor cells. The main contents of this paper are as follows: 1. A novel and simple method for preparing silicon-based mesoporous materials is proposed. The pore size structure of mesoporous materials is adjusted by changing the material ratio in the preparation process, and the mesoporous materials are modified organically by using specific materials to improve the macroscopic physical and chemical properties of mesoporous materials. The idea of mesoporous materials as a biological nano-container was proposed and applied to the biochemical reaction system. 2. A fluorescence analysis method for the detection of ATP by releasing fluorescein molecules from biological nano-containers was created. The complementary chain of the aptamer was combined to the surface of the nano-container, and the fluorescein molecule was enclosed in the nano-container by DNA hybrid gold nanoparticles. The ATP, aptamer was specifically combined with ATP molecule to make the gold nanoparticles leave the surface of the nano-container and release fluorescein molecules. The qualitative and quantitative analysis of ATP is realized by detecting fluorescence signal. The application of biological nano-container as drug carrier in cancer treatment was explored. 3. A novel method based on fluorescence detection technology was designed to detect telomerase by chain cycle amplification technique. The hairpin DNA was modified on the surface of magnetic nanoparticles, and the telomerase precursor was complementary to the partial sequence of hairpin DNA. The structure of hairpin DNA stem ring could be opened by the introduction of telomerase, and then a large number of fluorescent DNA and a small amount of biological nano-probes connected to DNA could be added to the system when the probe was fixed on the surface of magnetic nanoparticles under the action of DNA polymerase. The connected DNA began to grow and replaced the telomerase precursor to participate in the next cycle, which played an obvious role in amplifying the signal detection. Because magnetic nanoparticles can carry multiple biological nanoprobes and gold nanoparticles can also carry multiple fluorescent DNA molecules, the whole experiment realizes multiple amplification of signals and has high sensitivity to telomerase detection.
【學位授予單位】：青島科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R730.4;O657.3
【參考文獻】
相關(guān)期刊論文 前1條
1 沈星燦,何錫文,梁宏;納米粒子特性與生物分析[J];分析化學;2003年07期
，本文編號：2498001
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