肿瘤细胞内活性物质的分析和检测
发布时间：2019-06-12 11:54
【摘要】：癌症,一种严重威胁着人类生命健康的疾病,随着人类生存环境的恶化,其发病率逐年升高。癌症患者的治愈率极低,所以癌症的治疗以及早期的侦测检查成为近些年来医学上的一大难题。越早的发现肿瘤的存在,癌症患者被治愈的希望就越高。肿瘤细胞内活性物质的检测对于癌症的预防和诊断治疗等有着重要意义。本文通过纳米技术、DNA生物传感器技术以及荧光分析检测技术为检测肿瘤细胞内活性物提供了新的检测手段和思路。本文主要内容包括以下三个方面：1、提出了一种新颖简单的制备硅基介孔材料的方法。通过改变在制备过程的物料比来调节介孔材料的孔径结构,采用特定物料来对介孔材料进行有机功能化改性,进而改善介孔材料的宏观物理化学性质。提出了将介孔材料作为一种生物纳米容器的设想并将其应用到生物化学反应体系当中。2、创造了一种利用生物纳米容器释放荧光素分子检测ATP的荧光分析检测方法。将适体的互补链结合到纳米容器的表面,通过DNA杂交金纳米粒子将荧光素分子封闭在纳米容器内部。加入ATP,适体与ATP分子特异性结合,使金纳米粒子离开纳米容器表面,释放出荧光素分子。通过检测荧光信号,来实现对ATP的定性定量分析。对于生物纳米容器作为药物载体在癌症治疗中的应用做了初步探索。3、设计了一种新颖的基于荧光检测技术的基础上通过链循环放大技术检测端粒酶的方法。将发卡DNA修饰在磁性纳米微球的表面，引入端粒酶前体与发卡DNA的部分序列互补配对,端粒酶的引入可以将发卡DNA茎环结构打开,然后将携载大量荧光DNA以及少量连接DNA的生物纳米探针加入至体系当,当探针固定在磁性纳米微球的表面时,在DNA聚合酶的作用下,连接DNA开始生长并将端粒酶前体顶替下来,参与到下一个循环当中,对信号的检测起到了明显的放大作用。由于磁性纳米微球可以携载多个生物纳米探针,同时金纳米粒子也可携载多个荧光DNA分子,所以整个实验对信号实现了多重放大,对端粒酶的检测也有着较高的灵敏度。
[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.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R730.4;O657.3
本文编号：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.
【学位授予单位】：青岛科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R730.4;O657.3
【参考文献】
相关期刊论文 前1条
1 沈星灿,何锡文,梁宏;纳米粒子特性与生物分析[J];分析化学;2003年07期
，本文编号：2498001
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