【摘要】： 目的鼻咽癌(Nasopharyngeal Carcinoma, NPC)是我國高發(fā)的頭頸部惡性腫瘤,通常與體內(nèi)某些基因突變導(dǎo)致細(xì)胞周期改變和抗凋亡機制有關(guān)。本研究探討靶向survivin基因的小分子干擾RNA (short interfering RNA, siRNA)對人鼻咽癌CNE2細(xì)胞的生長和抑制作用。 方法設(shè)計、合成Survivin編碼基因序列特異的小分子干擾RNA,通過脂質(zhì)體轉(zhuǎn)染鼻咽癌細(xì)胞CNE2來敲除Survivin的表達(dá)。用半定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(reverse transcriptase polymerase chain reaction, RT-PCR)和蛋白免疫印跡法(Western blotting)測定Survivin mRNA和蛋白的表達(dá)。用流式細(xì)胞術(shù)檢測Survivin siRNA對細(xì)胞周期和細(xì)胞凋亡的影響。用甲基噻唑基四唑來分析Survivin小干擾RNA對癌細(xì)胞的生長抑制作用。 結(jié)果Survivin編碼基因序列特異siRNA能有效而穩(wěn)定地抑制鼻咽癌CNE2中Survivin mRNA和蛋白的表達(dá)；下調(diào)Survivin可以在體外顯著抑制腫瘤細(xì)胞的生長,在轉(zhuǎn)染后24和48小時后,轉(zhuǎn)染Survivin特異性siRNA組生長抑制率顯著高于對照組,用特異性Survivin siRNA處理后,檢測到有效的GO/Gi期阻滯。 結(jié)論RNA干擾技術(shù)能有效抑制Survivin基因的表達(dá),降低鼻咽癌細(xì)胞的增殖能力,促進(jìn)鼻咽癌細(xì)胞凋亡。Survivin特異性siRNA作為一種治療鼻咽癌的新手段值得進(jìn)一步深入研究。
[Abstract]:Objective Nasopharyngeal carcinoma (NPC) (Nasopharyngeal Carcinoma, NPC) is a high incidence malignant tumor of head and neck in China, which is usually related to the cell cycle change and anti-apoptosis mechanism caused by some gene mutations in the body. The aim of this study was to investigate the effects of small interfering RNA (short interfering RNA, siRNA) targeting survivin gene on the growth and inhibition of human nasopharyngeal carcinoma (NPC) CNE2 cells. Methods Survivin encoding gene sequence specific small interfering RNA, was synthesized to knockout the expression of Survivin by transfection of liposome into nasopharyngeal carcinoma cell CNE2. Semi-quantitative reverse transcriptase polymerase chain reaction (reverse transcriptase polymerase chain reaction, RT-PCR) and Western blot (Western blotting) were used to detect the expression of Survivin mRNA and protein. The effect of Survivin siRNA on cell cycle and apoptosis was detected by flow cytometry. Methylthiazolyl tetrazole was used to analyze the inhibitory effect of Survivin small interfering RNA on the growth of cancer cells. Results Survivin coding gene sequence specific siRNA could effectively and stably inhibit the expression of Survivin mRNA and protein in nasopharyngeal carcinoma (NPC) CNE2. Down-regulation of Survivin could significantly inhibit the growth of tumor cells in vitro. 24 and 48 hours after transfection, the growth inhibition rate of Survivin specific siRNA group was significantly higher than that of control group. After treatment with specific Survivin siRNA, effective GO/Gi phase arrest was detected. Conclusion RNA interference technique can effectively inhibit the expression of Survivin gene, reduce the proliferation ability of nasopharyngeal carcinoma cells and promote the apoptosis of nasopharyngeal carcinoma cells. Survivin specific siRNA is a new method for the treatment of nasopharyngeal carcinoma.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.63

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 李娟;趙瑩;張殿寶;黃蓓暉;;Survivin在多發(fā)性骨髓瘤患者骨髓細(xì)胞中的表達(dá)及其臨床意義[J];癌癥;2005年12期

2 朱瑾,徐如君,吳正虎,閆春蘭,曹曉林,方瑜,林琳;喉鱗狀細(xì)胞癌組織中Survivin基因與p15和p16基因表達(dá)的關(guān)系[J];中華耳鼻咽喉科雜志;2004年06期

3 徐澍,張維元;Ⅲ期鼻咽癌p53蛋白表達(dá)與生物學(xué)行為及預(yù)后的關(guān)系及其與p53基因突變、潛伏膜蛋白-1表達(dá)的相關(guān)研究[J];中華病理學(xué)雜志;2001年06期

4 李剛;李湘平;劉雄;彭英;林李家宓;;DNA載體介導(dǎo)RNA干擾抑制LMP-1基因?qū)Ρ茄拾┘?xì)胞轉(zhuǎn)移能力的影響[J];中華腫瘤雜志;2006年10期

，

本文編號：2388460