GANT61诱导食管鳞癌细胞凋亡的作用及其机制

王雷; 杜媛鲲; 王林; 米源; 廖海江

doi:10.3971/j.issn.1000-8578.2016.08.002

GANT61诱导食管鳞癌细胞凋亡的作用及其机制

Effect of GANT61 on Apoptosis of Esophageal Squamous Carcinoma Cells and Its Mechanism

摘要

摘要:
目的研究Gli抑制剂GANT61诱导人食管鳞癌细胞OE21和KYSE-30凋亡的机制。
方法应用不同浓度（0、10、15 μmol/L）的GANT61处理OE21和KYSE-30细胞24 h后, Western blot观察GANT61作用于OE21和KYSE-30细胞后对Gli1、Gli2、Fas和Bcl-2蛋白表达的影响。流式细胞术观察细胞凋亡及Fas、Bcl-2蛋白的表达。
结果 Western blot显示GANT61可以剂量依赖性显著下调OE21和KYSE-30细胞Gli1、Gli2和Bcl-2蛋白表达, 增加Fas蛋白表达。流式细胞术检测显示不同浓度GANT61作用24 h后, OE21和KYSE-30细胞表现出不同程度的细胞凋亡并且呈药物剂量依赖性, 与空白对照组相比差异均有统计学意义（P=0.000）；与空白对照组相比, OE21和KYSE-30细胞中Bcl-2蛋白表达水平显著降低且呈药物剂量依赖性, 差异均有统计学意义（P=0.000）； Fas蛋白表达水平显著增加且呈药物剂量依赖性, 差异均有统计学意义（P=0.000）。
结论 GANT61明显诱导细胞凋亡, 其作用机制可能与其通过特异性抑制OE21和KYSE-30细胞中Gli1、Gli2表达从而调节Fas和Bcl-2蛋白表达有关。

Abstract:
Objective To investigate the effect of GANT61 on the apoptosis of esophageal squamous carcinoma cells OE21 and KYSE-30, and to explore its possible mechanism.
Methods After OE21 and KYSE-30 cell lines were treated with different concentrations（0, 10, 15μmol/L）of GANT61 for 24h, the protein expression of Gli1, Gli2, Fas and bcl-2 were detected by Western blot. Cell apoptosis, Fas and bcl-2 expression levels of OE21 and KYSE-30 cell lines were measured by flow cytometry.
Results Western blot assay showed that GANT61 decreased Gli1, Gli2, Bcl-2 protein expression and increased Fas protein expression in a dose-dependent manner. Compared with DMSO group, the apoptotic rates of OE21 and KYSE-30 cell lines were significantly increased in a dose-dependent manner(P=0.000); the Bcl-2 protein expression levels were significantly decreased in a dose-dependent manner(P=0.000); the Fas protein expression levels were significantly increased in a dose-dependent manner(P=0.000).
Conclusion GANT61 could enhance the apoptotic rates of OE21 and KYSE-30 cells, and this effect may be associated with GANT61 decreasing Gli1 and Gli2 expression to regulate Fas and Bcl-2 protein expression levels in esophageal squamous carcinoma cells.

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