MicroRNA-1246与宫颈癌发生机制的相关性研究

发布时间：2018-05-02 23:42

本文选题：miR-1246 + 宫颈癌　；参考：《中南大学》2014年硕士论文

【摘要】：宫颈癌是严重影响妇女健康的一种恶性肿瘤,其发病率仅次于乳腺癌。其主要的发病原因是持续性的高危型人乳头状瘤病毒(HPV)感染。HPVE6早期调控基因主要与HPV的致癌性相关,编码其重要的致癌蛋白。MicroRNAs是一类长度为17-25nt的非编码RNA分子,通过与下游靶基因的3’端非翻译区结合,抑制翻译过程,从而调控蛋白编码基因的转录后翻译。近年来的研究表明microRNAs广泛参与包括癌症在内的各种疾病发生发展,可以作为疾病的诊断、预后的分子标志以及潜在的治疗靶点。MicroRNAs可以通过影响癌细胞的增殖、迁移、侵袭、细胞周期与凋亡等过程参与癌症的发生发展过程。MicroRNA-1246(miR-1246)定位于人2号染色体上,其表达受抑癌蛋白p53调控,参与唐氏综合症、系统性红斑狼疮等的发病过程。本文主要对miR-1246与宫颈癌发生机制的相关性进行研究。本文首先利用实时荧光定量PCR的方法,比较了68例宫颈癌组织及52例对照样本的miR。1246的表达水平,分析了miR-1246的表达水平与临床病理特征的关系。结果表明：宫颈癌组织中的miR-1246的表达水平显著低于对照组(p0.05),miR-1246的低表达与临床分期相关(p0.05),与年龄、肿瘤直径、宫颈浸润深度、淋巴结转移、脉管浸润无关(p0.05)。由于发现miR-1246在HPV16感染的宫颈癌组织及细胞系中比在HPV16未感染的宫颈癌组织及细胞系中的表达水平低。因此,我们利用过表达和(或)基因沉默癌细胞系(株)中的HPV16E6等方法,探讨了HPV16E6致癌蛋白与miR-1246表达之间的关系。结果表明：HPV16(+)宫颈癌细胞系SiHa中,敲除掉HPV16E6后,miR-1246表达上调,其下游靶蛋白DYRK1A表达降低；而HPV16(-)宫颈癌细胞系C33A转染HPV16E6后,miR-1246表达下调,其下游靶蛋白DYRK1A表达升高。表明HPV16E6致癌蛋白对miR-1246有明显的负调控作用。本文进一步采用模拟体内成熟miR-1246及siRNA干扰内源性miR-1246等方法研究了miR-1246对C33A细胞增殖、迁移、细胞周期的影响。结果发现miR-1246对宫颈癌细胞系C33A的增殖和细胞周期无显著影响,但对其细胞迁移能力有一定影响。
[Abstract]:Cervical cancer is a malignant tumor that seriously affects women's health, its incidence is second only to breast cancer. The main cause is the persistent high risk HPV infection. The early regulatory gene of HPVE6 is mainly related to the carcinogenicity of HPV. MicroRNAs, an important carcinogenic protein, are a class of non-coding RNA molecules with length of 17-25nt. By binding to the 3 '-terminal untranslated region of the downstream target gene, the process of translation was inhibited and the post-transcriptional translation of the protein encoding gene was regulated. Recent studies have shown that microRNAs is widely involved in the occurrence and development of various diseases, including cancer, and can be used as a molecular marker of disease diagnosis, prognosis and potential therapeutic target. MicroRNAs can affect the proliferation, migration and invasion of cancer cells. Cell cycle and apoptosis. MicroRNA-1246 miR-1246) is located on human chromosome 2. Its expression is regulated by tumor suppressor protein p53 and participates in the pathogenesis of Down's syndrome and systemic lupus erythematosus. The relationship between miR-1246 and the mechanism of cervical cancer was studied in this paper. The expression of miR.1246 in 68 cases of cervical carcinoma and 52 cases of control were compared by real-time fluorescence quantitative PCR. The relationship between the expression of miR-1246 and clinicopathological features was analyzed. The results showed that the expression of miR-1246 in cervical carcinoma was significantly lower than that in control group. The low expression of miR-1246 was correlated with clinical stage, age, tumor diameter, depth of cervix invasion, lymph node metastasis and vascular invasion. It was found that the expression of miR-1246 in HPV16 infected cervical cancer tissues and cell lines was lower than that in HPV16 uninfected cervical cancer tissues and cell lines. Therefore, we studied the relationship between HPV16E6 oncoprotein and miR-1246 expression by using overexpression and / or gene silencing of HPV16E6 in cancer cell lines (lines). The results showed that the expression of miR-1246 was up-regulated and the expression of downstream target protein DYRK1A was decreased after knockout of HPV16E6 in the cervical cancer cell line SiHa, while the expression of miR-1246 was down-regulated and the expression of DYRK1A in the downstream target protein was increased after transfection of C33A into the cervical cancer cell line C33A. The results showed that HPV16E6 oncoprotein had a negative effect on miR-1246. The effects of miR-1246 on the proliferation, migration and cell cycle of C33A cells were further studied by simulating mature miR-1246 in vivo and interfering with endogenous miR-1246 by siRNA. The results showed that miR-1246 had no significant effect on the proliferation and cell cycle of cervical cancer cell line C33A, but had a certain effect on the migration ability of cervical cancer cell line C33A.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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