微小RNA-34a在喉鱗狀細胞癌的功能研究及臨床意義
本文關(guān)鍵詞: 微小RNA-34a 生存素 喉鱗狀細胞癌 抑癌因子 出處:《寧波大學》2012年碩士論文 論文類型:學位論文
【摘要】:目的 生存素(Survivin)具有強大的抗凋亡作用,被認為是腫瘤基因治療的理想靶點。微小RNA-34a (miR-34a)通過負向調(diào)控某些腫瘤中特定基因的表達而發(fā)揮抑癌因子的作用。因此,本研究旨在探討miR-34a和Survivin在喉鱗狀細胞癌(LSCC)中相互作用的關(guān)系。 方法 1.采用脂質(zhì)體把miR-34a類似物導(dǎo)入人喉鱗狀細胞癌細胞系Hep-2細胞中,以提高Hep-2細胞中miR-34a的表達水平。四甲基偶氮唑藍(MTT)法檢測Hep-2細胞的增殖情況,,蛋白免疫印跡(Western blot)法檢測miR-34a對Survivin蛋白表達的影響,流式細胞術(shù)檢測細胞周期的變化。 2.實時定量聚合酶鏈反應(yīng)(PCR)檢測喉鱗狀細胞癌中miR-34a及Survivin信使RNA的表達水平,并分析其表達水平與喉鱗狀細胞癌的病理分化,淋巴結(jié)轉(zhuǎn)移,臨床分期及生存率的關(guān)系。 結(jié)果 1. miR-34a可以明顯的抑制Hep-2細胞的增殖,并且可以在不影響Survivin信使RNA的表達水平的情況下明顯下調(diào)Survivin蛋白的表達。此外,miR-34a使Hep-2細胞的細胞周期阻滯在G0/G1期。 2.喉鱗狀細胞癌組織標本結(jié)果表明,miR-34a在腫瘤組織中是低表達的,而Survivin是高表達的。miR-34a的表達水平與喉鱗狀細胞癌患者的病理分化,淋巴結(jié)轉(zhuǎn)移,臨床分期負相關(guān),與生存率正相關(guān)。 結(jié)論 miR-34a可能通過靶向Survivin與喉鱗狀細胞癌的發(fā)生密切相關(guān),在喉鱗狀細胞癌的發(fā)生發(fā)展中起著重要作用。
[Abstract]:Purpose Survivin (survivin) has a strong anti-apoptotic effect. Small RNA-34a miR-34a) acts as a tumor suppressor by negatively regulating the expression of specific genes in certain tumors. The purpose of this study was to investigate the interaction between miR-34a and Survivin in laryngeal squamous cell carcinoma (LSCC). Method 1. MiR-34a analogue was introduced into human laryngeal squamous cell carcinoma cell line Hep-2 cells by liposome. In order to increase the expression of miR-34a in Hep-2 cells, the proliferation of Hep-2 cells was detected by MTT assay. The effect of miR-34a on the expression of Survivin protein was detected by Western blotting, and the cell cycle was detected by flow cytometry. 2.Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of miR-34a and Survivin messenger RNA in laryngeal squamous cell carcinoma. The relationship between the expression level and pathological differentiation, lymph node metastasis, clinical stage and survival rate of laryngeal squamous cell carcinoma (LSCC) was analyzed. Results 1. MiR-34a could significantly inhibit the proliferation of Hep-2 cells. Moreover, the expression of Survivin protein could be significantly down-regulated without affecting the expression level of Survivin messenger RNA. MiR-34a blocked the cell cycle of Hep-2 cells in G _ 0 / G _ 1 phase. 2. The expression of miR-34a in laryngeal squamous cell carcinoma tissues was low. However, the expression of .miR-34a in Survivin was negatively correlated with pathological differentiation, lymph node metastasis, clinical stage and survival rate in laryngeal squamous cell carcinoma. Conclusion MiR-34a may be closely related to laryngeal squamous cell carcinoma by targeting Survivin, and play an important role in the development of laryngeal squamous cell carcinoma.
【學位授予單位】:寧波大學
【學位級別】:碩士
【學位授予年份】:2012
【分類號】:R739.65
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