【摘要】： 目的鼻咽癌(Nasopharyngeal Carcinoma, NPC)是我國高發(fā)的頭頸部惡性腫瘤,通常與體內某些基因突變導致細胞周期改變和抗凋亡機制有關。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細胞的生長和抑制作用。 方法設計、合成Survivin編碼基因序列特異的小分子干擾RNA,通過脂質體轉染鼻咽癌細胞CNE2來敲除Survivin的表達。用半定量逆轉錄聚合酶鏈反應(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。用流式細胞術檢測Survivin siRNA對細胞周期和細胞凋亡的影響。用甲基噻唑基四唑來分析Survivin小干擾RNA對癌細胞的生長抑制作用。 結果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；下調Survivin可以在體外顯著抑制腫瘤細胞的生長,在轉染后24和48小時后,轉染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。 結論RNA干擾技術能有效抑制Survivin基因的表達,降低鼻咽癌細胞的增殖能力,促進鼻咽癌細胞凋亡。Survivin特異性siRNA作為一種治療鼻咽癌的新手段值得進一步深入研究。
[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.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R739.63

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