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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 effect of dachshund family transcription factor 1 on the proliferation and cell cycle of human endometrial carcinoma HEC-1A cells in vitro. Methods: The DACH1-overexpressed recombinant lentivirus LV-DACH1-GFP was transfected into human endometrial carcinoma HEC-1A cells. The localization of DACH1 in HEC-1A cells was observed by laser confocal microscopy. The expression of DACH1 was detected by real-time fluorescence quantitative PCR The expression of DACH1 mRNA and protein in HEC-1A cells was detected by Western blotting. The proliferation of HEC-1A cells in vitro was detected by MTT assay. The cell cycle distribution was detected by FCM. Results: LV-DACH1-GFP was mainly distributed in the nucleus after being infected with recombinant lentivirus LV-DACH1-GFP in human endometrial carcinoma HEC-1A cells. The expression levels of DACH1 m RNA and protein in HEC-1A cells in LV-DACH1-GFP infected group were significantly higher than those in control group (HEC-1A cells infected with lentivirus LV-GFP) and blank group (HEC-1A cells without any (P <0.01). The proliferation of HEC-1A cells in LV-DACH1-GFP group was significantly inhibited (P <0.05), and the cell cycle was arrested in G0 / G1 phase (P <0.01). Conclusion: D ACH1 gene can inhibit the proliferation of human endometrial carcinoma HEC-1A cells, which may be related to the cell cycle arrest of DACH1 gene.