实时荧光定量RT-PCR检测鼻咽癌Survivin mRNA基因表达

Detection of Survivin mRNA in nasopharyngeal carcinoma by real-time fluorescence quantitative RT-PCR

目的建立实时荧光定量RT-PCR方法检测鼻咽癌组织中Survivin mRNA的表达。方法用Trizol方法提取鼻咽癌CNE-2细胞株和鼻咽癌组织总RNA后,将其逆转录为cDNA,建立实时荧光定量RT-PCR方法,检测人鼻咽癌Survivin mRNA的表达,以慢性鼻咽炎患者鼻咽部组织作为对照。结果鼻咽癌细胞株、鼻咽癌组和慢性鼻咽炎组患者鼻咽部组织均可检测到Survivin基因mRNA的表达,Survivin mRNA在鼻咽癌组织中的表达较慢性鼻咽炎患者鼻咽部组织高,两者差异有显著性意义(P<0.01)。Ⅲ+Ⅳ期和Ⅰ+Ⅱ期鼻咽癌患者均检测到Survivin基因mRNA的表达,但相对表达量无统计学意义。Survivin mRNA表达与鼻咽癌患者的年龄、性别无相关性。结论实时荧光定量RT-PCR方法是一种快速有效灵敏度高的鼻咽癌Survivin mRNA表达的定量检测方法,Survivin mRNA的过度表达可能在鼻咽癌发生、发展过程中起一定作用。

Objective To establish the method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in nasopharyngeal carcinoma （NPC） tissues. Methods The total RNA was extracted from NPC cell line CNE-2 and NPC tissues with Tfizol and then been transcribed reversely to cDNA. A method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin rnRNA in NPC tissues was established, in which chronic nasopharyngitis tissues were regarded as control group. Results The expression of Survivin mRNA all could been detected either in CNE-2 cells, NPC tissues or in chronic nasopharyngitis tissues, and the expression of Survivin mRNA in NPC tissues was higher than that in chronic tissues, the difference was significantly（ P 〈 0.01 ） .The expression of Survivin mRNA both could been detected in Ⅰ ＋ Ⅱ and Ⅲ ＋ Ⅳ stage NPC, and the relative quantiflcations of gene expression in Ⅰ ＋ Ⅱ and Ⅲ ＋ Ⅳ stage NPC were undistinguished（ P 〉 0.05）. There was no significant difference between Survivin mRNA expression and age, sex of NPC patients （ P 〉 0.05）. Conclusion Real time fluorescence quantitative RT-PCR is a rapid, effective and high sensitivity method for detecting the expression of Survivin mRNA in NPC tissues. The overexpression of Survivin mRNA may play some roles in pathogenesis of NPC.