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.