Five percent of normal cystic fibrosis transmembrane conductance regulator mRNA ameliorates the severity of pulmonary disease in cystic fibrosis
Five percent of normal cystic fibrosis transmembrane conductance regulator mRNA ameliorates the severity of pulmonary disease in cystic fibrosis
复制标题
DOI：
10.1165/rcmb.2001-0004oc
复制
发表时间：
2002-11-01
影响因子：
6.4
通讯作者：
Amaral, MD
中科院分区：
文献类型：
--
作者：
Ramalho, AS;Beck, S;Amaral, MD
囊性纤维化(CF)跨膜电导调节因子(CFTR)基因转录本水平的估计范围为正常的4-20%。鉴于获得可靠数据对治疗策略的重要性，我们开发了一种新的基于聚合酶链式反应的方法来定量cftr转录本，并将其应用于分析5例CF患者和3272-26A>；G/F508del基因型的鼻黏膜上皮RNA。我们计算出在这5例患者中存在的总CFTRRNA的8.2+/-0.84%是正常的全长CFTRmRNA。然后，我们(在F508del携带者的鼻样中，n=30)证明了全长F508del CFTR转录本的丰度比野生型转录本低，并估计F508del/野生型转录本的平均比率为0.87+/-0.06。为了确定相对于正常个体发现的全长转录本的数量，我们校正了F508del转录本的较低丰度，并计算出五名CF患者的野生型CFTRmRNA平均水平为正常水平的4.7+/-0.45%。由于与F508del纯合子相比，这些患者有轻微的CF，这种CFTRmRNA水平似乎足以避免疾病的严重并发症。
Estimates of the level of transcripts from the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene required to develop a CF phenotype range from 4-20% of normal. Due to the importance of obtaining reliable data on this issue for therapeutic strategies, we developed a novel polymerise chain reaction-based method to quantify CFTR transcripts and applied it to the analysis of nasal epithelium RNA of five patients with CF and the 3272-26A>G/F508del genotype. We calculated that 8.2 +/- 0.84% of the total CFTR RNA present in these five patients is normal full-length CFTR mRNA. We then demonstrated (in nasal samples from F508del carriers, n = 30) that the abundance of full-length F508del CFTR transcripts is reduced compared with wild-type transcripts, and estimated that the average ratio of F508del/wild-type transcripts is 0.87 +/- 0.06. To determine the amount of full-length transcripts relative to levels found in normal individuals, we corrected for the lower abundance of the F508del transcripts and calculated that the five patients with CF have, on average, 4.7 +/- 0.45% of the normal level of wild-type CFTR mRNA. Because these patients have mild CF compared with F508del homozygotes, this CFTR mRNA level appears to be sufficient to avoid the severe complications of the disease.