SOX2在5-氟尿嘧啶处理的人喉癌Hep-2细胞中的表达及作用机制

发布时间：2018-04-03 12:24

本文选题：人喉癌　切入点：SOX　出处：《中国医科大学学报》2014年03期

【摘要】：目的探讨5-氟尿嘧啶(5-FU)对人喉癌Hep-2细胞中干细胞转录因子SOX2表达的影响及机制,并分析SOX2拮抗5-FU诱导的细胞凋亡作用。方法 CCK-8法检测48 h内不同浓度5-FU对Hep-2细胞生长的抑制作用,统计抑制率和IC50值。Western blot法检测5-FU作用不同时间后SOX2的表达量,并检测PI3K/AKT信号活化情况。Hoechst染色法检测细胞凋亡。结果5-FU对Hep-2细胞生长抑制作用明显,IC50值为20.92μg/mL。随着时间的延长SOX2表达增高,48 h达到最大值,磷酸化AKT活化也随着时间的延长逐渐增加。使用PI3K/AKT信号特异性抑制剂LY294002处理后,SOX2表达被下调;Hoechst染色法显示细胞凋亡数增多。同时检测凋亡相关蛋白表达变化,结果显示Survivin、Bcl-2、BAX在5-FU处理的Hep-2细胞中表达均上升,但PI3K/AKT信号抑制后抗凋亡蛋白Survivin、Bcl-2被显著下调,凋亡蛋白BAX表达继续上调。结论 PI3K/AKT信号活化诱导SOX2表达在Hep-2细胞拮抗5-FU诱导的细胞凋亡中发挥了重要作用。
[Abstract]:Objective to investigate the effect and mechanism of 5-fluorouracil (5-FU) on the expression of transcription factor SOX2 in human laryngeal carcinoma Hep-2 cells, and to analyze the effect of SOX2 on the apoptosis induced by 5-FU.Methods CCK-8 assay was used to detect the inhibitory effect of 5-FU at different concentrations on the growth of Hep-2 cells within 48 h. The expression of SOX2 was detected by IC50 value. Western blot assay was used to detect the expression of SOX2 and the activation of PI3K/AKT signal. Hoechst staining was used to detect the apoptosis of Hep-2 cells.Results the inhibitory effect of 5-FU on the growth of Hep-2 cells was found to be 20.92 渭 g / mL.The expression of SOX2 increased to the maximum at 48 h, and the activation of phosphorylated AKT increased with the prolongation of time.After treated with PI3K/AKT signal specific inhibitor LY294002, the expression of SOX2 was down-regulated by Hoechst staining.At the same time, the expression of apoptosis-related protein was detected. The results showed that the expression of survivin Bcl-2mBAX increased in 5-FU treated Hep-2 cells, but the anti-apoptotic protein survivin Bcl-2 was significantly down-regulated and the expression of apoptotic protein BAX continued to up-regulate after PI3K/AKT signal suppression.Conclusion SOX2 expression induced by PI3K/AKT signal activation plays an important role in Hep-2 cells antagonizing apoptosis induced by 5-FU.
【作者单位】：中国医科大学附属第一医院耳鼻咽喉科;
【基金】：辽宁省自然科学基金(201202287)
【分类号】：R739.65

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