Okamoto Y, Nakabayashi H, Kikukawa S, Nakano H
Department of Clinico-Laboratory Diagnostics, Nara Medical University, Kashihara, Japan.
Rinsho Byori. 1996 May;44(5):483-6.
Cyclin and cyclin-dependent kinase(cdk) complexes, and their inhibitors (CKIs) play important roles in growth regulation on the cells. p27/kip1 is a CKI associated with G1 arrest induced by cell to cell contact, transforming growth factor-beta and cyclic AMP. The abnormality of p27/Kip1 genes in human tumors usually appears as a steady level defect of expression, since mutations in them is rare. Thus it is important to estimate the expression level of this gene. To detect the change of p27/Kip1 mRNA level in blood cells, we developed the ribonuclease protection assay using nonradioactive riboprobe which was produced by reverse transcriptase-polymerase chain reaction (RT-PCR) with T7 promoter-added antisense primer and the in vitro transcription system. Our assay may be useful for clinical evaluation of the mRNA level.
细胞周期蛋白与细胞周期蛋白依赖性激酶（CDK）复合物及其抑制剂（CKIs）在细胞生长调控中发挥着重要作用。p27/kip1是一种与细胞间接触、转化生长因子-β和环磷酸腺苷诱导的G1期阻滞相关的CKI。人类肿瘤中p27/Kip1基因的异常通常表现为表达水平的稳定缺陷，因为它们的突变很少见。因此，评估该基因的表达水平很重要。为了检测血细胞中p27/Kip1 mRNA水平的变化，我们利用非放射性核糖探针开发了核糖核酸酶保护试验，该探针由添加了T7启动子的反义引物通过逆转录聚合酶链反应（RT-PCR）和体外转录系统产生。我们的试验可能有助于对mRNA水平进行临床评估。
