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目的研究华蟾素对体外培养的两株子宫内膜癌细胞(PTEN表达完整的HEC-1A细胞和PTEN缺失的Ishikawa细胞)生物学行为影响并探讨分子机制。方法以不同浓度华蟾素作用两株子宫内膜癌细胞,MTT和克隆形成法检测细胞增殖;流式细胞法检测细胞周期及凋亡率;Transwell法检测侵袭;RT-PCR和Western blot法检测胰岛素生长因子-1受体(IGF-1R)基因和蛋白表达。结果 PTEN基因缺失的Ishikawa细胞随华蟾素浓度升高,增殖和侵袭能力明显降低,凋亡率显著升高,G1期细胞比例明显增多,IGF-1R基因和蛋白表达显著下降,HEC-1A细胞对华蟾素作用不敏感。结论PTEN基因缺失可增强子宫内膜癌细胞对华蟾素的敏感性。
Objective To investigate the effects of cinobufacini on the biological behavior of two endometrial carcinoma cells (PTEN-expressing HEC-1A cells and PTEN-deficient Ishikawa cells) cultured in vitro and to explore the molecular mechanism. Methods The proliferation of two endometrial cancer cells treated with different concentration of cinobufacini was detected by MTT and clonogenic assay. The cell cycle and apoptosis rate were detected by flow cytometry. The invasion was detected by Transwell assay. The expression of catenin was detected by RT-PCR and Western blot Insulin growth factor-1 receptor (IGF-1R) gene and protein expression. Results The Ishikawa cells lacking PTEN gene had a significant increase in cinobufacini concentration, significantly decreased the proliferation and invasion ability, significantly increased the apoptosis rate, increased the proportion of cells in G1 phase and decreased the expression of IGF-1R gene and protein, The role of cinobufotalin insensitive. Conclusion PTEN gene deletion enhances the sensitivity of endometrial carcinoma cells to cinobufacini.