RNAi-Mediated Knock-Down of Arylamine N-acetyltransferase-1 Expression Induces E-cadherin Up-Regulation and Cell-Cell Contact Growth Inhibition
RNAi-Mediated Knock-Down of Arylamine N-acetyltransferase-1 Expression Induces E-cadherin Up-Regulation and Cell-Cell Contact Growth Inhibition
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DOI：
10.1371/journal.pone.0017031
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发表时间：
2011-02-09
期刊：
影响因子：
3.7
通讯作者：
Minchin, Rodney F.
中科院分区：
文献类型：
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作者：
Tiang, Jacky M.;Butcher, Neville J.;Minchin, Rodney F.
芳胺N-乙酰基转移酶-1(NAT1)是一种催化芳胺和肼底物生物转化的酶。它也在叶酸代谢物对氨基苯甲酰谷氨酸的分解代谢中起作用。最近的生物信息学研究已经将Nat1的表达与不同的癌症亚型相关联。然而，Nat1在细胞生物学中的直接作用还没有确定。在这项研究中，我们在结肠腺癌细胞系HT-29中敲除了Nat1，发现细胞形态发生了显著的变化，伴随着细胞间接触生长抑制的增加和细胞融合时细胞活力的丧失。NAT1基因的敲除也导致软琼脂中锚定独立生长的减弱。NAT1的缺失导致E-钙粘蛋白的mRNA和蛋白水平上调。E-钙粘附素的这种变化并不是由于RNAi的非靶点效应，在前列腺癌细胞系22Rv1中也观察到了这种变化。在体内，与稳定转RNAi的细胞或亲本的HT-29细胞相比，Nat1基因敲除的细胞生长有更长的倍增时间。这项研究表明，Nat1影响细胞的生长和形态。此外，该研究还表明，Nat1可能是癌症治疗的一个新的药物靶点。
Arylamine N-acetyltransferase-1 (NAT1) is an enzyme that catalyzes the biotransformation of arylamine and hydrazine substrates. It also has a role in the catabolism of the folate metabolite p-aminobenzoyl glutamate. Recent bioinformatics studies have correlated NAT1 expression with various cancer subtypes. However, a direct role for NAT1 in cell biology has not been established. In this study, we have knocked down NAT1 in the colon adenocarcinoma cell-line HT-29 and found a marked change in cell morphology that was accompanied by an increase in cell-cell contact growth inhibition and a loss of cell viability at confluence. NAT1 knock-down also led to attenuation in anchorage independent growth in soft agar. Loss of NAT1 led to the up-regulation of E-cadherin mRNA and protein levels. This change in E-cadherin was not attributed to RNAi off-target effects and was also observed in the prostate cancer cell-line 22Rv1. In vivo, NAT1 knock-down cells grew with a longer doubling time compared to cells stably transfected with a scrambled RNAi or to parental HT-29 cells. This study has shown that NAT1 affects cell growth and morphology. In addition, it suggests that NAT1 may be a novel drug target for cancer therapeutics.