DACH1基因对人子宫内膜癌HEC-1A细胞增殖的影响

Effect of DACH 1 gene expression on proliferation of human endometrial carcinoma HEC-1A cells

目的:探讨细胞命运决定因子DACH1(dachshund family transcription factor 1)对人子宫内膜癌HEC-1A细胞体外增殖能力和细胞周期的影响。方法 :将DACH1过表达的重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞,在激光共聚焦显微镜下观察DACH1在HEC-1A细胞中的定位;分别采用实时荧光定量PCR法和蛋白质印迹法检测HEC-1A细胞中DACH1 m RNA和蛋白的表达,MTT法检测HEC-1A细胞的体外的增殖能力,FCM法检测细胞周期的分布。结果 :重组慢病毒LV-DACH1-GFP感染人子宫内膜癌HEC-1A细胞后,LV-DACH1-GFP主要分布在细胞核中。LV-DACH1-GFP感染组HEC-1A细胞中,DACH1 m RNA和蛋白的表达水平均明显高于对照组(HEC-1A细胞感染慢病毒LV-GFP)和空白组(HEC-1A细胞未进行任何感染)(P值均<0.01),LV-DACH1-GFP感染组HEC-1A细胞的增殖受到明显抑制(P<0.05),细胞周期阻滞于G0/G1期(P<0.01)。结论 :D ACH1基因可抑制人子宫内膜癌HEC-1A细胞的增殖,这一作用可能与DACH1基因阻滞细胞周期有关。

Objective： To investigate the effects of cell fate determination factor dachshund family transcription factor 1 （DACH1） on proliferation and cell cycle of human endometrial carcinoma HEC-1A cells. Methods： The recombinant lentivirus LV-DACH1-GFP with DACH1 overexpression was infected into human endometrial carcinoma HEC- 1A cells. The location of DACH1 in HEC-1A cells was observed under a laser confocal microscope. The expressions of DACH1 mRNA and protein in HEC-1A cells after infection with LV-DACH1-GFP were detected by real-time fluorescent quantitative-PCR and Western blotting, respectively. The proliferation and cell cycle distribution were determined by MTT and FCM, respectively.Results： LV-DACH1-GFP expression was mainly located in nucleus of HEC-1A cells. The expression levels of DACH1 mRNA and protein in LV-DACH1-GFP infection group were higher than those in control group （HEC-1A cells infected with lentivirus LV-GFP） and blank group （HEC-1A cells without any infection） （all P 〈 0.01）. The proliferation of HEC-1A cells in LV-DACH1-GFP infection group was inhibited （P 〈 0.05）, and the cell cycle was arrested at Go/G1 phase （P 〈 0.01）.Conclusion： The inhibition effect of DACH1 gene on the proliferation of HEC-1A cells may be related to inducing cell cycle arrest.