本文選題：microRNA-195 + 宮頸癌��； 參考：《安徽醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:宮頸癌被認(rèn)為是婦科最常見的惡性腫瘤之一,同時嚴(yán)重危害婦女的身體健康。研究表明微小RNA(micro RNA,miRNA)與宮頸癌的發(fā)生發(fā)展有著密切的聯(lián)系。本文通過研究宮頸癌組織中miRNA-195(miR-195)的表達(dá)特點,以明確作用機(jī)制。方法:利用qRT-PCR方法分別檢測人宮頸癌組織與正常組織的miR-195的表達(dá)并進(jìn)行比較。培養(yǎng)宮頸癌細(xì)胞,并轉(zhuǎn)染miR-195以及其抑制物,采用MTT法、transwell小室法觀察miR-195對宮頸癌細(xì)胞在細(xì)胞增殖、細(xì)胞遷移以及細(xì)胞侵襲方面的作用。篩選miR-195可能的靶基因并采用western blot法觀察其表達(dá)及與miR-195的關(guān)系。結(jié)果:miR-195的表達(dá)在宮頸癌組織、細(xì)胞中均顯著地下調(diào)。過表達(dá)miR-195能顯著抑制細(xì)胞的生長。而抑制了miR-195則對細(xì)胞增殖具有相反的作用。miR-195的異常表達(dá)顯著地抑制了He La和C33A細(xì)胞的遷移。而阻斷miR-195的表達(dá)能增強(qiáng)He La和C33A細(xì)胞的遷移。過表達(dá)miR-195減輕了He La和C33A細(xì)胞的侵襲,而阻斷miR-195的表達(dá)能增強(qiáng)He La和C33A細(xì)胞的侵襲。根據(jù)生物信息學(xué)分析預(yù)測,HDGF是miR-195的直接靶基因。根據(jù)western blot結(jié)果,在宮頸癌細(xì)胞中,HDGF顯著上調(diào)。在He La和C33A細(xì)胞轉(zhuǎn)染了miR-195 mimics以后,HDGF在m RNA水平不受影響;相反的,轉(zhuǎn)染了miR-195 mimics以后,HDGF在蛋白水平下調(diào),而轉(zhuǎn)染了miR-195 inhibitor以后,HDGF在蛋白水平顯著上調(diào)。結(jié)論:miR-195在細(xì)胞試驗中抑制了宮頸癌細(xì)胞的增殖,遷移以及侵襲。確定了HDGF是miR-195的m RNA直接靶點。miR-195通過靶向調(diào)控HDGF進(jìn)而發(fā)揮抑制宮頸癌細(xì)胞增殖,遷移以及侵襲行為。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R737.33

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