FOXA 1对人乳头状甲状腺癌细胞增殖与细胞周期的影响及其机制探讨

摘要

Objective To observe the effect of forkhead box protein A1 (FOXA1 )on the proliferation and cell cycle of human papillary thyroid carcinoma TPC1 cells and to investigate its mechanism.Methods TPC1 cells were divided into two groups which were separately transfected with Flag-vector (empty vector group)and Flag-FOXA1 (FOXA1 group). The proliferation activity of the two groups of TPC1 cells was detected by MTT method after transfection for 12 h,24 h and 36 h,and cell cycle was detected by flow cytometry at 24 h.The mRNA and protein expression of p27 Kip1 ,cyclin-de-pendent kinase (CDK2)and CyclinD1 was detected by real-time PCR and Western blotting at 24 h after transfection of Flag-FOXA1 .Results In the FOXA1 group,the cell proliferation was promoted,cells in the G1 phase decreased,but cells in the S phase increased.Compared with the empty vector group,the expression of p27 Kip1 mRNA and protein was decreased and the expression of CyclinD1 and CDK2 mRNA and protein was increased.Conclusions FOXA1 can pro-mote the cell proliferation and increase the cells in the S period.The mechanism may be related to the down-regulation of p27 Kip1 and up-regulation of CDK2 and CyclinD1 expression.%目的：观察叉头框蛋白A1（FOXA1）对人乳头状甲状腺癌TPC1细胞增殖及细胞周期的影响，并探讨其机制。方法培养TPC1细胞，分为两组，分别转染Flag-vector（空载体组）及Flag-FOXA1（FOXA1组）。转染12、24、36 h时，采用MTT法检测两组细胞增殖活性。转染24 h后，采用流式单染细胞术检测细胞周期，采用Real-time PCR法检测细胞周期调控蛋白27（p27 Kip1）、细胞周期蛋白依赖性激酶2（CDK2）、细胞周期蛋白D1（Cyclin D1） mRNA的表达，采用Western blot法检测p27 Kip1、CDK2、Cyclin D1蛋白的表达。结果与空载体组比较，FOXA1组细胞增殖率增加，G1期细胞比例减少，而 S 期细胞比例增加（P均＜0．05）。与空载体组比较，FOXA1组 p27 Kip1 mRNA及蛋白表达降低，CDK2、Cyclin D1 mRNA及蛋白表达增加（P均＜0．05）。结论 FOXA1可促进TPC1细胞增殖，使S期细胞增加，其机制可能与下调p27 Kip1及上调CDK2、CyclinD1表达有关。