microRNA-195通过靶基因HDGF抑制宫颈癌的肿瘤行为研究

发布时间：2018-05-15 13:00

本文选题：microRNA-195 + 宫颈癌　；参考：《安徽医科大学》2016年博士论文

【摘要】：目的:宫颈癌被认为是妇科最常见的恶性肿瘤之一,同时严重危害妇女的身体健康。研究表明微小RNA(micro RNA,miRNA)与宫颈癌的发生发展有着密切的联系。本文通过研究宫颈癌组织中miRNA-195(miR-195)的表达特点,以明确作用机制。方法:利用qRT-PCR方法分别检测人宫颈癌组织与正常组织的miR-195的表达并进行比较。培养宫颈癌细胞,并转染miR-195以及其抑制物,采用MTT法、transwell小室法观察miR-195对宫颈癌细胞在细胞增殖、细胞迁移以及细胞侵袭方面的作用。筛选miR-195可能的靶基因并采用western blot法观察其表达及与miR-195的关系。结果:miR-195的表达在宫颈癌组织、细胞中均显著地下调。过表达miR-195能显著抑制细胞的生长。而抑制了miR-195则对细胞增殖具有相反的作用。miR-195的异常表达显著地抑制了He La和C33A细胞的迁移。而阻断miR-195的表达能增强He La和C33A细胞的迁移。过表达miR-195减轻了He La和C33A细胞的侵袭,而阻断miR-195的表达能增强He La和C33A细胞的侵袭。根据生物信息学分析预测,HDGF是miR-195的直接靶基因。根据western blot结果,在宫颈癌细胞中,HDGF显著上调。在He La和C33A细胞转染了miR-195 mimics以后,HDGF在m RNA水平不受影响;相反的,转染了miR-195 mimics以后,HDGF在蛋白水平下调,而转染了miR-195 inhibitor以后,HDGF在蛋白水平显著上调。结论:miR-195在细胞试验中抑制了宫颈癌细胞的增殖,迁移以及侵袭。确定了HDGF是miR-195的m RNA直接靶点。miR-195通过靶向调控HDGF进而发挥抑制宫颈癌细胞增殖,迁移以及侵袭行为。
[Abstract]:Objective: cervical cancer is considered to be one of the most common malignant tumors in gynecology. Studies have shown that RNA(micro miRNAs are closely related to the occurrence and development of cervical cancer. The expression of miRNA-195 miR-195 in cervical carcinoma was studied to clarify the mechanism. Methods: qRT-PCR method was used to detect the expression of miR-195 in human cervical carcinoma and normal tissues. Cervical cancer cells were cultured and transfected with miR-195 and their inhibitors. The effects of miR-195 on cell proliferation, cell migration and cell invasion were observed by MTT transwell chamber assay. The possible target genes of miR-195 were screened and their expression and relationship with miR-195 were observed by western blot assay. Results the expression of miR-195 was significantly down-regulated in cervical cancer cells. Overexpression of miR-195 significantly inhibited cell growth. On the contrary, inhibition of miR-195 on cell proliferation. The abnormal expression of miR-195 significantly inhibited the migration of He-La and C33A cells. Blocking the expression of miR-195 enhanced the migration of He-La and C33A cells. Overexpression of miR-195 alleviated the invasion of He-La and C33A cells, while blocking the expression of miR-195 enhanced the invasion of He-La and C33A cells. According to bioinformatics analysis, it is predicted that HDGF is a direct target gene of miR-195. According to western blot results, HDGF was significantly up-regulated in cervical cancer cells. After transfection of He-La and C33A cells into miR-195 mimics, the level of m RNA was not affected. On the contrary, after transfection of miR-195 mimics, the protein level of He-La and C33A cells was down-regulated, but after transfection of miR-195 inhibitor, the level of miR-195 mimics was significantly up-regulated. Conclusion the cell proliferation, migration and invasion of cervical cancer cells were inhibited by the cell test. It is confirmed that HDGF is the direct target of m RNA of miR-195. MiR-195 inhibits the proliferation, migration and invasion of cervical cancer cells by targeting HDGF.
【学位授予单位】：安徽医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R737.33

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