结直肠癌组织定量RT-PCR分析中内参基因的选择

Selection of internal reference genes for real-time quantitative RT-PCR of colorectal cancer tissue

目的:筛选适用于结直肠癌组织样本的内源性参照基因。方法:以10例结直肠癌患者组织总RNA为研究材料,采用实时定量RT-PCR检测GAPDH、ACTINB、UBC、HRPT1和18S r RNA这5个候选内参基因的表达量,应用ge Norm和Norm Finder两种软件分析选择最佳内参基因。结果:ge Norm软件发现GAPDH与ACTINB的M值相对最小,为最佳内参基因组合,18S r RNA的M值最大,稳定度最低;而Norm Finder软件分析认为GAPDH表达稳定性最佳,UBC次之。结论:适用于结直肠癌组织样本RT-PCR实验的内参基因为GAPDH,最佳组合为GAPDH与ACTINB,而18S r RNA及HRPT1不是合适的内参基因。

Objective：To select the optimal internal reference genes for gene expression analysis in colorectal tumor tissues.Methods：The total RNA of colorectal cancer tissues from 10 patients weve studied. The m RNA transcription profiles of five frequently used housekeeping genes,including GAPDH,ACTINB,UBC,HRPT1,and 18 S r RNA,were tested by real-time quantitative RT-PCR.The stability of these control genes was analyzed by ge Norm and Norm Finder softwares. Results：The M value of GAPDH and ACTINB was the least and identified as the suitable and stable internal reference gene combinations using ge Norm software. The M volue of18 S r RNA was the highest and its stability was the lowest. Furthermore,GAPDH was the most stable reference gene,which was followed by UBC using Norm Finder software. Conclusion：The combination of GAPDH and ACTINB,but not 18 S r RNA and HRPT1,is the most reliable reference gene group for normalization of real-time RT-PCR in colorectal tumor tissues.