结直肠癌组织定量RT-PCR分析中内参基因的选择
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国家自然科学基金资助(81373091);江苏省高校优势学科建设资助(公共卫生与预防医学)


Selection of internal reference genes for real-time quantitative RT-PCR of colorectal cancer tissue
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    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 rRNA这5个候选内参基因的表达量,应用geNorm和NormFinder两种软件分析选择最佳内参基因-结果:geNorm软件发现GAPDH与ACTINB的M值相对最小,为最佳内参基因组合,18S rRNA的M值最大,稳定度最低;而NormFinder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 rRNA及HRPT1不是合适的内参基因-

    Abstract:

    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 mRNA transcription profiles of five frequently used housekeeping genes,including GAPDH,ACTINB,UBC,HRPT1,and 18S rRNA,were tested by real-time quantitative RT-PCR. The stability of these control genes was analyzed by geNorm and NormFinder softwares. Results:The M value of GAPDH and ACTINB was the least and identified as the suitable and stable internal reference gene combinations using geNorm software. The M volue of 18S rRNA was the highest and its stability was the lowest. Furthermore,GAPDH was the most stable reference gene,which was followed by UBC using NormFinder software. Conclusion:The combination of GAPDH and ACTINB,but not 18S rRNA and HRPT1,is the most reliable reference gene group for normalization of real-time RT-PCR in colorectal tumor tissues.

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黎书炜,杜牧龙,金 经,储海燕,朱陵君,张正东,王美林.结直肠癌组织定量RT-PCR分析中内参基因的选择[J].南京医科大学学报(自然科学版),2016,(8):952-955

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  • 收稿日期:2016-01-23
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  • 在线发布日期: 2016-08-19
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