發(fā)布時(shí)間：2018-02-06 07:24

  本文關(guān)鍵詞： 腎癌 miR-216b-5p 表達(dá) 功能　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:腎細(xì)胞癌(renal cell carcinoma,RCC)簡(jiǎn)稱腎癌,是泌尿生殖系統(tǒng)中最常見的惡性腫瘤之一,也是腎臟實(shí)質(zhì)中最常見的惡性腫瘤,其惡性程度較高。由于腎癌患者早期癥狀和體征往往不明顯,缺乏特異性的臨床表現(xiàn),因此早期診斷較為困難,以致約三分之一的腎癌患者在明確診斷時(shí)腫瘤已經(jīng)發(fā)生了遠(yuǎn)處臟器轉(zhuǎn)移。由于腎癌對(duì)放療和普通化療不敏感,手術(shù)切除病灶成為有效的治療方法,但是仍有20-30%的腎癌患者在接受手術(shù)治療后出現(xiàn)了腫瘤復(fù)發(fā)。因此,研究腎癌發(fā)生發(fā)展的作用機(jī)制并尋找腎癌早期診斷、療效評(píng)價(jià)、預(yù)后評(píng)估的分子標(biāo)志物就顯得尤為重要。微小RNA(micro RNA或者miRNA)是一類具有大約21-23個(gè)核苷酸長(zhǎng)度的短鏈非編碼RNA分子,它主要通過與靶基因的m RNA 3’端非編碼區(qū)(3'-UTR區(qū))一些特異性序列結(jié)合,從而降解靶基因m RNA或者抑制其翻譯,在轉(zhuǎn)錄后水平起到調(diào)控基因表達(dá)的作用。目前的研究表明,miRNA在包括腎癌在內(nèi)的多種腫瘤組織中異常表達(dá),并參與細(xì)胞增殖、侵襲、遷移和凋亡等生命過程之中。在此我們以miR-216b-5p和腎癌作為研究對(duì)象,研究miR-216b-5p在腎癌發(fā)生發(fā)展過程中的功能及相關(guān)致癌機(jī)制,為后續(xù)開展miRNA在腎癌診斷和治療中的研究提供一些理論基礎(chǔ)。方法:應(yīng)用qRT-PCR方法檢測(cè)41例腎癌組織和配對(duì)癌旁正常組織中miR-216b-5p的表達(dá)水平,并使用統(tǒng)計(jì)學(xué)軟件分析miR-216b-5p在腎癌組織中的表達(dá)水平與患者臨床病理特征之間的相關(guān)性。然后分別通過CCK-8增殖實(shí)驗(yàn)、細(xì)胞劃痕實(shí)驗(yàn)和流式細(xì)胞術(shù)分別檢測(cè)miR-216b-5p對(duì)腎癌細(xì)胞增殖、遷移和凋亡的影響。最后通過生物信息學(xué)方法預(yù)測(cè)miR-216b-5p的潛在靶基因。結(jié)果:與配對(duì)癌旁正常組織相比,miR-216b-5p在腎癌組織中的表達(dá)水平明顯降低(P0.05),而且其在腎癌組織中的表達(dá)水平與患者性別、年齡、病理類型、TNM分期、臨床AJCC分期之間無(wú)明顯相關(guān)性(P0.05)。通過轉(zhuǎn)染miR-216b-5pmimics上調(diào)腎癌細(xì)胞中miR-216b-5p的表達(dá)水平后,與對(duì)照組相比,細(xì)胞增殖和遷移能力受到抑制,同時(shí)促進(jìn)了細(xì)胞凋亡(P0.05)。結(jié)論:miR-216b-5p在腎癌組織中表達(dá)下調(diào),并通過抑制腎癌細(xì)胞增殖和遷移,促進(jìn)腎癌細(xì)胞凋亡,從而發(fā)揮抑癌基因作用。KRAS可能是miR-216b-5p的其中一個(gè)靶基因。
[Abstract]:Objective: renal cell carcinoma (RCC) is one of the most common malignant tumors in the genitourinary system. It is also the most common malignant tumor in renal parenchyma, its malignant degree is high. Because the early symptoms and signs of renal carcinoma patients are often not obvious, lack of specific clinical manifestations, so the early diagnosis is more difficult. As a result, about 1/3 of RCC patients had distant organ metastasis at the time of definite diagnosis. Because RCC is insensitive to radiotherapy and chemotherapy, surgical resection of tumor has become an effective treatment method. However, there are still 20-30% of patients with renal cell carcinoma have recurrence after surgery. Therefore, to study the mechanism of the development of renal cell carcinoma and to search for early diagnosis of renal cell carcinoma, evaluation of curative effect. Molecular markers for prognostic evaluation are particularly important. Small RNA(micro RNA or miRNAs. It is a class of short chain noncoding RNA molecules with about 21-23 nucleotide lengths. It degrades the target gene m RNA or inhibits its translation by binding to some specific sequences of the target gene in the 3'-terminal non-coding region of the target gene. At the post-transcriptional level, it plays a role in regulating gene expression. Current studies have shown that miRNA is abnormal expressed in various tumor tissues, including renal cell carcinoma, and participates in cell proliferation and invasion. In the process of migration and apoptosis, we focus on miR-216b-5p and renal cell carcinoma (RCC). To study the function of miR-216b-5p in the development of renal cell carcinoma and its carcinogenic mechanism. To provide a theoretical basis for the further study of miRNA in the diagnosis and treatment of renal cell carcinoma. Methods:. QRT-PCR method was used to detect the expression of miR-216b-5p in 41 cases of renal cell carcinoma and adjacent normal tissues. Statistical software was used to analyze the correlation between the expression level of miR-216b-5p and the clinicopathological characteristics of RCC. Then CCK-8 proliferation test was used. Cell scratch assay and flow cytometry were used to detect the proliferation of RCC cells by miR-216b-5p. The effects of migration and apoptosis. Finally, the potential target genes of miR-216b-5p were predicted by bioinformatics. Results: compared with matched adjacent normal tissues. The expression level of miR-216b-5p in RCC was significantly lower than that in RCC, and its expression level was correlated with sex, age and pathological type of RCC. There was no significant correlation between clinical AJCC staging and P0.05.After transfection of miR-216b-5pmimics, the expression of miR-216b-5p in RCC cells was up-regulated. Compared with the control group, cell proliferation and migration were inhibited and apoptosis was promoted by P0.05.Conclusion the expression of micromiR-216b-5p is down-regulated in renal cell carcinoma. KRAs may be one of the target genes of miR-216b-5p by inhibiting the proliferation and migration of RCC cells and promoting the apoptosis of RCC cells.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.11

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相關(guān)期刊論文 前1條

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本文編號(hào)：1493906