应用siRNA技术下调Survivin抑制鼻咽癌细胞生长的研究
发布时间：2018-12-20 21:56
【摘要】： 目的鼻咽癌(Nasopharyngeal Carcinoma, NPC)是我国高发的头颈部恶性肿瘤,通常与体内某些基因突变导致细胞周期改变和抗凋亡机制有关。本研究探讨靶向survivin基因的小分子干扰RNA (short interfering RNA, siRNA)对人鼻咽癌CNE2细胞的生长和抑制作用。 方法设计、合成Survivin编码基因序列特异的小分子干扰RNA,通过脂质体转染鼻咽癌细胞CNE2来敲除Survivin的表达。用半定量逆转录聚合酶链反应(reverse transcriptase polymerase chain reaction, RT-PCR)和蛋白免疫印迹法(Western blotting)测定Survivin mRNA和蛋白的表达。用流式细胞术检测Survivin siRNA对细胞周期和细胞凋亡的影响。用甲基噻唑基四唑来分析Survivin小干扰RNA对癌细胞的生长抑制作用。 结果Survivin编码基因序列特异siRNA能有效而稳定地抑制鼻咽癌CNE2中Survivin mRNA和蛋白的表达；下调Survivin可以在体外显著抑制肿瘤细胞的生长,在转染后24和48小时后,转染Survivin特异性siRNA组生长抑制率显著高于对照组,用特异性Survivin siRNA处理后,检测到有效的GO/Gi期阻滞。 结论RNA干扰技术能有效抑制Survivin基因的表达,降低鼻咽癌细胞的增殖能力,促进鼻咽癌细胞凋亡。Survivin特异性siRNA作为一种治疗鼻咽癌的新手段值得进一步深入研究。
[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.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
本文编号：2388460
[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.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
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