胃癌中E2F3相关调控网络的分析及miR-320a，let-7d对其调控作用的研究
发布时间：2019-05-28 11:52
【摘要】：20世纪以来,胃癌这一恶性肿瘤发病率极高,世界范围内胃癌患者病死率仅次于肺癌位居世界第二位。据统计,截止2016年,70%以上的胃癌患者来自于发展中国家,侧面说明了胃癌发病与空气、污染、水源、人口数量等生存环境息息相关。目前胃癌晚期患者的五年生存率仍然较低,因此进一步探究胃癌细胞浸润生长模式以及胃癌发生转移精确的分子机制,筛选出胃癌诊断过程中易于检测的关键靶点,对于胃癌的预防和治疗具有十分重要的意义。转录因子E2F3在促进癌症转移进程方面具有十分重要的作用,精确构建E2F3基因相关的调控网络可以帮助人们深入了解胃癌各个时期发生发展的机制。Micro RNA与相应靶基因结合调控基因的转录,在肿瘤的各个时期发挥了重要作用。本研究旨在系统、全面的解析胃癌中E2F3相关的转录调控网络的表达模式,并进一步探讨miR-320a和let-7d对E2F3表达水平的影响。研究通过基因芯片技术筛选胃癌组织中差异表达明显的基因和micro RNA,结合生物信息学软件预测E2F3相关的micro RNA、转录因子以及靶基因,构建以转录因子E2F3为中心的转录调控网络,结合大量文献富集分析调控网络中E2F3与相关micro RNA之间的调控关系。生物信息学分析表明E2F3可能是miR-320a和let-7d的靶基因,通过实验进一步验证miR-320a和let-7d与E2F3之间的调控关系。第一部分胃癌中TF-micro RNA转录调控网络的构建通过分析10对胃癌及癌旁正常组织的差异基因表达谱和10对micro RNA表达谱芯片数据,我们筛选到809个上调和92个下调的差异表达基因,58个上调和8个下调的差异表达micro RNA。综合TRED数据库筛选,其中共有32个差异基因与E2F家族调控相关,以差异基因为研究对象,进行功能显著性分析和信号通路分析,此部分研究为寻找胃癌相关致病基因提供了数据信息及理论依据。结合Targetscan等micro RNA数据库的预测,筛选出miR-320a,miR-200,miR-150,miR-125a-5p,miR-92a,miR-31,miR-30,miR-17,let-7d,let-7i等10个micro RNA与E2F3相关。经过大量文献富集分析,我们推测miR-320a和let-7d极有可能通过调控E2F3从而影响胃癌的转移分化进程。第二部分miR-320a和let-7d对E2F3调控作用的研究通过qRT-PCR及Western blot检测转录因子E2F3和miR-320a、let-7d在胃癌中的表达水平,发现E2F3在胃癌组织中表达量升高,其表达水平与胃癌的分化程度,临床分期等因素显著相关;miR-320a,let-7d在胃癌组织和细胞中显著低表达,其表达水平与E2F3的表达有明显的相关性。使用miR-320a和let-7d mimics转染SGC-7901及MGC-803细胞系验证过表达miR-320a和let-7d对E2F3的影响,结果显示在miR-320a和let-7d过表达细胞中E2F3的表达水平显著降低,表明E2F3的表达与miR-320a,let-7d的表达水平呈显著负相关,由此可见miR-320a和let-7d可能通过调控转录因子E2F3的表达进而影响胃癌的转移分化进程。
[Abstract]:Since the 20th century, the incidence of gastric cancer is very high, and the mortality of gastric cancer patients ranks second only to lung cancer in the world. According to statistics, up to 2016, more than 70% of patients with gastric cancer come from developing countries, which shows that the incidence of gastric cancer is closely related to air, pollution, water source, population and other living environment. At present, the 5-year survival rate of patients with advanced gastric cancer is still low, so we further explore the infiltration and growth pattern of gastric cancer cells and the accurate molecular mechanism of metastasis of gastric cancer, and screen out the key targets that are easy to detect in the diagnosis of gastric cancer. It is of great significance for the prevention and treatment of gastric cancer. Transcription factor E2F3 plays a very important role in promoting cancer metastasis. The accurate construction of E2F3 gene-related regulatory network can help people to understand the mechanism of gastric cancer occurrence and development. Micro RNA and the transcription of corresponding target gene binding regulatory genes, and play an important role in each stage of tumor. The purpose of this study was to systematically and comprehensively analyze the expression patterns of E2F3-related transcriptional regulatory networks in gastric cancer and to further explore the effects of miR-320a and let-7d on the expression of E2F3. In this study, gene chip technique was used to screen differentially expressed genes in gastric cancer tissues and micro RNA, combined with bioinformatics software to predict micro RNA, transcription factors and target genes related to E2F3, and a transcriptional regulatory network centered on transcription factor E2F3 was constructed. Combined with a large number of literature enrichment, the regulatory relationship between E2F3 and related micro RNA in the regulatory network is analyzed. Bioinformatics analysis showed that E2F3 may be the target gene of miR-320a and let-7d. The regulatory relationship between miR-320a and let-7d and E2F3 was further verified by experiments. In the first part, the construction of TF-micro RNA transcriptional regulatory network in gastric cancer was carried out by analyzing the differential gene expression profiles of 10 pairs of gastric cancer and adjacent normal tissues and the microarray data of 10 pairs of micro RNA expression profiles. We screened 809 up-regulated and 92 down-regulated differentially expressed genes, 58 up-regulated and 8 down-regulated differentially expressed micro RNA.. A total of 32 differential genes were related to the regulation of E2F family by comprehensive TRED database screening. The functional significance analysis and signal pathway analysis were carried out with the difference gene as the research object. This part of the study provides data information and theoretical basis for searching for gastric cancer related pathogenic genes. Combined with the prediction of micro RNA database such as Targetscan, 10 micro RNA, such as miR-320a,miR-200,miR-150,miR-125a-5p,miR-92a,miR-31,miR-30,miR-17,let-7d,let-7i, are selected to be related to E2F3. After a large number of literature enrichment analysis, we speculate that miR-320a and let-7d may affect the metastasis and differentiation of gastric cancer by regulating E2F3. The second part is the study on the regulatory effect of miR-320a and let-7d on E2F3. The expression of transcription factors E2F3 and miR-320a,let-7d in gastric cancer was detected by qRT-PCR and Western blot. It was found that the expression of E2F3 was increased in gastric cancer. The expression level of gastric cancer was significantly correlated with the degree of differentiation and clinical stage of gastric cancer. The expression of miR-320a,let-7d was significantly low in gastric cancer tissues and cells, and its expression level was significantly correlated with the expression of E2F3. SGC-7901 and MGC-803 cell lines were transformed with miR-320a and let-7d mimics to verify the effect of overexpression of miR-320a and let-7d on E2F3. The results showed that the expression level of E2F3 in miR-320a and let-7d overexpressed cells was significantly decreased. The results showed that the expression of E2F3 was negatively correlated with the expression of miR-320a,let-7d. It can be seen that miR-320a and let-7d may affect the metastasis and differentiation of gastric cancer by regulating the expression of transcription factor E2F3.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2
本文编号：2487028
[Abstract]:Since the 20th century, the incidence of gastric cancer is very high, and the mortality of gastric cancer patients ranks second only to lung cancer in the world. According to statistics, up to 2016, more than 70% of patients with gastric cancer come from developing countries, which shows that the incidence of gastric cancer is closely related to air, pollution, water source, population and other living environment. At present, the 5-year survival rate of patients with advanced gastric cancer is still low, so we further explore the infiltration and growth pattern of gastric cancer cells and the accurate molecular mechanism of metastasis of gastric cancer, and screen out the key targets that are easy to detect in the diagnosis of gastric cancer. It is of great significance for the prevention and treatment of gastric cancer. Transcription factor E2F3 plays a very important role in promoting cancer metastasis. The accurate construction of E2F3 gene-related regulatory network can help people to understand the mechanism of gastric cancer occurrence and development. Micro RNA and the transcription of corresponding target gene binding regulatory genes, and play an important role in each stage of tumor. The purpose of this study was to systematically and comprehensively analyze the expression patterns of E2F3-related transcriptional regulatory networks in gastric cancer and to further explore the effects of miR-320a and let-7d on the expression of E2F3. In this study, gene chip technique was used to screen differentially expressed genes in gastric cancer tissues and micro RNA, combined with bioinformatics software to predict micro RNA, transcription factors and target genes related to E2F3, and a transcriptional regulatory network centered on transcription factor E2F3 was constructed. Combined with a large number of literature enrichment, the regulatory relationship between E2F3 and related micro RNA in the regulatory network is analyzed. Bioinformatics analysis showed that E2F3 may be the target gene of miR-320a and let-7d. The regulatory relationship between miR-320a and let-7d and E2F3 was further verified by experiments. In the first part, the construction of TF-micro RNA transcriptional regulatory network in gastric cancer was carried out by analyzing the differential gene expression profiles of 10 pairs of gastric cancer and adjacent normal tissues and the microarray data of 10 pairs of micro RNA expression profiles. We screened 809 up-regulated and 92 down-regulated differentially expressed genes, 58 up-regulated and 8 down-regulated differentially expressed micro RNA.. A total of 32 differential genes were related to the regulation of E2F family by comprehensive TRED database screening. The functional significance analysis and signal pathway analysis were carried out with the difference gene as the research object. This part of the study provides data information and theoretical basis for searching for gastric cancer related pathogenic genes. Combined with the prediction of micro RNA database such as Targetscan, 10 micro RNA, such as miR-320a,miR-200,miR-150,miR-125a-5p,miR-92a,miR-31,miR-30,miR-17,let-7d,let-7i, are selected to be related to E2F3. After a large number of literature enrichment analysis, we speculate that miR-320a and let-7d may affect the metastasis and differentiation of gastric cancer by regulating E2F3. The second part is the study on the regulatory effect of miR-320a and let-7d on E2F3. The expression of transcription factors E2F3 and miR-320a,let-7d in gastric cancer was detected by qRT-PCR and Western blot. It was found that the expression of E2F3 was increased in gastric cancer. The expression level of gastric cancer was significantly correlated with the degree of differentiation and clinical stage of gastric cancer. The expression of miR-320a,let-7d was significantly low in gastric cancer tissues and cells, and its expression level was significantly correlated with the expression of E2F3. SGC-7901 and MGC-803 cell lines were transformed with miR-320a and let-7d mimics to verify the effect of overexpression of miR-320a and let-7d on E2F3. The results showed that the expression level of E2F3 in miR-320a and let-7d overexpressed cells was significantly decreased. The results showed that the expression of E2F3 was negatively correlated with the expression of miR-320a,let-7d. It can be seen that miR-320a and let-7d may affect the metastasis and differentiation of gastric cancer by regulating the expression of transcription factor E2F3.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2
【参考文献】
相关期刊论文 前2条
1 尹娜,王新娟,尚永丰;中介因子复合体在基因转录调控中的作用[J];生理科学进展;2003年04期
2 ;Gene expression profiles of hepatoma cell line HLE[J];World Journal of Gastroenterology;2003年04期
，本文编号：2487028
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2487028.html
