杜氏盐藻14-3-3基因抑制食管鳞癌细胞凋亡的研究
首发时间:2008-12-22
摘要:目的:探讨杜氏盐藻14-3-3基因在食管鳞癌细胞株EC-9706凋亡中的作用。方法:构建真核表达载体pEGFP-C1-14-3-3和pcDNA3.1(+)-14-3-3,脂质体方法转染食管鳞癌细胞株EC-9706,荧光显微镜观察EGFP-14-3-3融合蛋白在细胞中的定位;G-418筛选pcDNA3.1(+) -14-3-3稳定细胞株,RT-PCR和Western blotting法检测转染细胞株中14-3-3基因的表达;Western blotting检测转染前后EC-9706细胞中Bcl-2蛋白的表达变化。结果:EGFP-14-3-3融合蛋白定位于细胞核;获得pcDNA3.1(+)-14-3-3稳定转染细胞株,且转染后细胞中Bcl-2蛋白表达量明显增高。结论:杜氏盐藻14-3-3基因具有抑制食管鳞癌细胞凋亡的作用。
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Inhibitory Effects of the 14-3-3 gene of Dunaliella salina on apoptosis in esophageal squamous cancer cells
Abstract:Aim: To investigate the effect of the 14-3-3 gene of Dunaliella salina on apoptosis of esophageal squamous cell cancer cell line EC-9706. Methods: The fragment of 14-3-3 was inserted into the corresponding restriction sites of the eukaryotic expression vector pEGFP-C1 and pcDNA3.1 (+), respectively, to obtain recombinant eukaryotic expression vectors pEGFP-C1-14-3-3 and pcDNA3.1(+)-14-3-3. The recombinant vectors were transfected into EC-9706 cells by LipofectamineTM 2000. The location of EGFP-14-3-3 proteins was determined by fluorescent microscope. After EC-9706 cells integrated with pcDNA3.1(+)-14-3-3 were selected, expression of the 14-3-3 gene of Dunaliella salina was detected by RT-PCR and Western blotting. The effect of 14-3-3 on expression of Bcl-2 was detected by Western blotting. Results: EGFP-14-3-3 proteins were localized in the nucleus of EC-9706 cells. EC-9706 cells stably expressing the 14-3-3 gene of Dunaliella salina were obtained. The expression of Bcl-2 protein was obviously increased in the EC-9706 cells transfected with pcDNA3.1(+)-14-3-3, compared to the control groups. Conclusions: It is concluded that the 14-3-3 gene of Dunaliella salina inhibits the apoptosis of the EC-9706 cells.
Keywords: Dunaliella salina 14-3-3 gene Esophageal squamous cell cancer Apoptosis
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