Triptolide enhances the sensitivity of gastric cancer BGC823 cells to 5-FU by suppressing TLR4 expression
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摘要: 目的 观察雷公藤内酯醇(TPL)是否可通过抑制Toll样受体4(TLR4)表达以增强胃癌BGC823细胞对5-FU的敏感性，并分析其相关的下游分子机制。方法 采用CCK-8法检测不同浓度TPL以及TPL联合5-FU对BGC823细胞的增殖的影响。将BGC823细胞分为对照组、5-FU组(仅予5-FU)、TPL+5-FU组以及脂多糖(LPS)+TPL+5-FU组；Hoechst33258染色观察细胞凋亡形态学变化，流式细胞仪检测细胞凋亡率，Western blot检测TLR4、磷酸化AKT(p-AKT)、Survivin、活性Caspase-3蛋白表达。结果 TPL可呈浓度-时间依赖性的抑制BGC823细胞增殖。与5-FU组比较，TPL联合5-FU后对BGC823细胞的抑制率明显增加(P< 0.01)，其5-FU的半数抑制浓度(IC50)显著降低(P< 0.01)。与5-FU组比较，TPL+5-FU组凋亡细胞数和细胞凋亡率均显著增加(P< 0.01)；而LPS+TPL+5-FU组凋亡细胞数以及细胞凋亡率均较TPL+5-FU组显著下降(P< 0.05)。TPL+5-FU组的TLR4、p-AKT、Survivin蛋白表达均较5-FU组下降，而活性Caspase-3蛋白表达增加；与TPL+5-FU组比较，LPS+TPL+5-FU组的p-AKT、Survivin蛋白表达增加，活性Caspase-3蛋白表达减少。结论 TPL具有增强BGC823细胞对5-FU敏感性的作用，其机制与抑制TLR4表达，进而阻断PI3K/AKT通路、下调Survivin表达并促进Caspase-3活化有关。
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关键词:
- 雷公藤内酯醇 /
- Toll样受体4 /
- 胃癌 /
- 磷酸化AKT /
- 活性Caspase-3
Abstract: Objective To observe whether triptolide (TPL) could enhance the sensitivity of gastric cancer BGC823 cells to 5-FU by inhibiting the expression of Toll like receptor 4 (TLR4), and analyze the related downstream molecular mechanisms.Methods The CCK-8 method was used to detect the effects of different concentrations of TPL and TPL combined with 5-FU on the proliferation of BGC823 cells. BGC823 cells were divided into control group, 5-FU group, TPL+5-FU group and lipopolysaccharide (LPS) +TPL+5-FU group; Hoechst33258 staining was used to observe the apoptosis morphological changes and flow cytometry was used to detect the cell apoptosis rate. TLR4, phosphorylated Akt (p-Akt), Survivin and Caspase-3 protein expression were detected by Western blot.Results TPL could inhibit the proliferation of BGC823 cells in a concentration-time dependent manner. Compared with the 5-FU group, the inhibition rate of the TPL combined with 5-FU group on BGC823 cells increased significantly (P< 0.01), and the half inhibitory concentration (IC50) of 5-FU decreased significantly (P< 0.01). Compared with the 5-FU group, the number of apoptotic cells and apoptosis rate of the TPL+5-FU group were significantly increased(P< 0.01); However, the number of apoptotic cells and cell apoptosis rate of the LPS+TPL+5-FU group were significantly lower than those of the TPL+5-FU group (P< 0.05). The TLR4, p-Akt and Survivin protein expression of the TPL+5-FU group were higher than those of the 5-FU group, and the expression of active Caspase-3 protein increased. Compared with the TPL+5-FU group, the p-Akt and Survivin protein expression of the LPS+TPL+5-FU group increased, but active Caspase-3 protein expression reduced.Conclusion TPL can enhance the sensitivity of BGC823 cells to 5-FU, its mechanism is related to inhibiting TLR4 expression, blocking the PI3K/AKT pathway, down regulating Survivin expression and promoting Caspase-3 activation.-
Key words:
- triptolide /
- Toll like receptor 4 /
- gastric cancer /
- phosphorylated AKT /
- active Caspase-3
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