CAJ | 학술논문

　　目的研究人肝再生增强因子（ALR）对细胞周期素依赖性激酶1启动子（CDK1p）转录调节的作用，探讨ALR的生物学作用机制。方法根据GenBank中CDK1p序列的分析设计引物，应用聚合酶链反应（PCR）扩增人CDK1基因启动子基因序列，并克隆至真核报告载体pCAT3-Basic中，构建CDK1启动子报告基因表达载体pCAT3-CDK1p，以该质粒转染HepG2细胞系，并同时与pcDNA3.1（－）-ALR共转染HepG2细胞系，用酶联免疫吸附法（ELISA）检测报告基因氯霉素乙酰转移酶（CAT）的表达活性。结果 pCAT3-CDK1p组CAT表达活性明显高于pCAT3-basic组，差异有统计学意义，pCAT3-CDK1p与pcDNA3.1（－）-ALR 共转染组的CAT表达活性明显低于pCAT3-CDK1p组，差异有统计学意义。结论 CDK1启动子有顺式激活下游基因的活性，ALR对CDK1基因有下调作用。
　　목적연구인간재생증강인자（ALR）대세포주기소의뢰성격매1계동자（CDK1p）전록조절적작용，탐토ALR적생물학작용궤제。방법근거GenBank중CDK1p서렬적분석설계인물，응용취합매련반응（PCR）확증인CDK1기인계동자기인서렬，병극륭지진핵보고재체pCAT3-Basic중，구건CDK1계동자보고기인표체재체pCAT3-CDK1p，이해질립전염HepG2세포계，병동시여pcDNA3.1（－）-ALR공전염HepG2세포계，용매련면역흡부법（ELISA）검측보고기인록매소을선전이매（CAT）적표체활성。결과 pCAT3-CDK1p조CAT표체활성명현고우pCAT3-basic조，차이유통계학의의，pCAT3-CDK1p여pcDNA3.1（－）-ALR 공전염조적CAT표체활성명현저우pCAT3-CDK1p조，차이유통계학의의。결론 CDK1계동자유순식격활하유기인적활성，ALR대CDK1기인유하조작용。
Objective To investigate the effects of human augmenter of liver regeneration (ALR) on the transcription of cyclin-dependent kinase 1 gene promoter (CDK1p), and to explore the biological function mechanism of ALR. Methods Polymerase chain reaction (PCR) technique was employed to amplify the sequence of CDK1 promoter from HepG2 genomic DNA, named CDK1p, and the PCR product was cloned into pCAT3-basic, named pCAT3-CDK1p. The HepG2 cells were transfected with pCAT3-CDK1p, and then co-transfected with pCAT3-CDK1p and pcDNA3.1(-)-ALR. The chloramphenicol acetyltransferase (CAT) activity was detected by an enzyme linked immunosorbent assay (ELISA) kit. Results The CAT activity of the HepG2 transfected by pCAT3-CDK1p was significantly higher than that of the negative control group. The CAT activity of the HepG2 co-transfected by pCAT3-CDK1p and pcDNA3.1(-)-ALR was significantly lower than that of the HepG2 transfected by pCAT3-CDK1p. Conclusions The CDK1p has transcription activity. It is suggested that ALR can down-regulate CDK1 gene.