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首页> 外文期刊>Surgery >A del T poly T (8) mutation in the 3' untranslated region (UTR) of the CDK2-AP1 gene is functionally significant causing decreased mRNA stability resulting in decreased CDK2-AP1 expression in human microsatellite unstable (MSI) colorectal cancer (CRC
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A del T poly T (8) mutation in the 3' untranslated region (UTR) of the CDK2-AP1 gene is functionally significant causing decreased mRNA stability resulting in decreased CDK2-AP1 expression in human microsatellite unstable (MSI) colorectal cancer (CRC

机译：CDK2-AP1基因3'非翻译区（UTR）中的del T poly T（8）突变在功能上很重要，导致mRNA稳定性下降，导致人类微卫星不稳定（MSI）大肠直肠癌（CRC）的CDK2-AP1表达下降

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摘要

BACKGROUND: We have previously published results indicating that decreased expression of CDK2-AP1 in MSI human colorectal cancer is associated with deletion mutations in the poly (T) 8 repeat sequence within the 3'-UTR of the CDK2-AP1 gene. In this study, we test the hypothesis that the del T mutation results in decreased CDK2-AP1 expression by causing reduced mRNA stability. METHODS: We introduced wild-type and mutant 3'-UTR sequences fused to a green fluorescent protein (GFP) gene separately into human CRC cell lines and quantified the expression of the GFP gene. Native CDK2-AP1 mRNA stability was measured in human CRC cell lines, using an actinomycin D assay and the mRNA structure folding software mfold 3.2. RESULTS: Mutant GFP-3'-UTR samples demonstrated significantly reduced GFP expression compared with wild-type GFP-3'-UTR as measured by both FACS and real-time PCR. Both the actinomycin D assay and mfold software demonstrated significantly reduced mRNA stability for the del T poly (T) 8 transcript compared with the wild type. CONCLUSIONS: In summary, these novel results support our hypotheses that the del T poly (T) 8 observed in the 3'-UTR of the CDK2-AP1 gene in human MSI CRC is functionally significant and results in decreased CDK2-AP1 expression. The results also indicate the mechanism of this decreased expression is caused at least in part by decreased mRNA stability.
机译：背景：我们先前已发表的结果表明，CDK2-AP1在MSI人大肠癌中的表达降低与CDK2-AP1基因3'-UTR内的poly（T）8重复序列的缺失突变有关。在这项研究中，我们测试了一个假说，即del T突变通过导致mRNA稳定性降低而导致CDK2-AP1表达降低。方法：我们将与绿色荧光蛋白（GFP）基因融合的野生型和突变型3'-UTR序列分别引入人CRC细胞系，并定量了GFP基因的表达。使用放线菌素D测定法和mRNA结构折叠软件mfold 3.2在人CRC细胞系中测量了天然CDK2-AP1 mRNA的稳定性。结果：通过FACS和实时PCR检测，与野生型GFP-3'-UTR相比，突变的GFP-3'-UTR样品显示GFP的表达显着降低。放线菌素D分析法和mfold软件均证明与野生型相比，del T poly（T）8转录本的mRNA稳定性大大降低。结论：总之，这些新颖的结果支持我们的假设，即在人MSI CRC的CDK2-AP1基因3'-UTR中观察到的del T poly（T）8在功能上很重要，并导致CDK2-AP1表达降低。结果还表明这种表达降低的机制至少部分地由降低的mRNA稳定性引起。

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