摘要
目的研究TUSC3基因在正常胰腺和胰腺癌组织中的表达水平。方法利用寡核苷酸基因芯片技术和实时定量PCR技术检测20例胰腺癌组织和6例正常胰腺组织中TUSC3基因的表达。芯片杂交结果进行Ratio值分析,实时定量PCR进一步验证杂交结果,PCR产物的定量方法采用比较Ct法。结果芯片扫描图像信号清晰,具有较低的整体背景和较高的信噪比。与正常胰腺组织相比,TUSC3基因在胰腺癌组织中表达下调,平均Ratio值为0.297±0.38。荧光定量PCR结果:TUSC3基因平均模板量在正常胰腺组织中为0.0387493±0.00965,胰腺癌组织中为0.0206028±0.00401;与正常胰腺组织相比,胰腺癌组织中TUSC3基因表达下调1.88倍(P<0.05)。结论TUSC3基因在胰腺癌组织中表达水平明显下调,其可能机制与该基因启动子区过甲基化有关,TUSC3基因在胰腺癌发生发展中的生物学效应值得进一步研究。
Objective To investigate the expression of the TUSC3 gene in human normal pancreas and pancreatic adenocarcinoma tissues. Methods The expression of TUSC3 gene in 20 cases of pancreatic cancer and 6 cases of normal pancreas were detected by oligonucleotide microarray and real-time quantitative PCR. Total RNA were extracted from each sample with TRIzol method. To assure the quality of obtained total RNA, three different examinations were performed which included spectrophotometer, agarose gel electrophoresis and Lab on chip examination. The results of mieroarray hybridization were analysised with Ratio value and verified by realtime quantitative PCR. The product of real-time quantitative PCR was quantitated by comparative Ct method. Results The scanning signal of microarray hybridization was clear, and the aquired images had a lower background and higher signal-noise ratio. The expression of TUSC3 gene in pancreatic adenocarcinoma against normal pancreas was down-regulated （ Cy3/Cy5 〈 0. 5 ） by Ratio analysis, which the average value of Ratio was 0. 297 ± 0. 38. The average quantity of TUSC3 gene in pancreatic adenocarcinoma and normal pancreas detected by QRT-PCR were 0. 0206028 ± 0. 00401 and 0. 0387493 ± 0. 00965 respectively. In comparison with normal pancreas, the expression of TUSC3 gene in pancreatic adenocarcinoma was down-expressed as 1.88 folds （ P 〈 0. 05 ）. Conclusion TUSC3 gene was significantly down-expressed in pancreatic adenocarcinoma and its mechanism might be related with age-related hypermethylation in promoter region. However, the real role of TUSC3 gene in pancreatic adenocarcinoma developing has to be evaluated in the future.
出处
《肝胆外科杂志》
2008年第6期463-465,共3页
Journal of Hepatobiliary Surgery
