热休克蛋白70抑制剂PES对人胃癌BGC细胞NF-κB通路的影响

Effect of heat shock protein 70 inhibitor PES on NF-κB signal pathway of human gastric cancer BGC cells

目的:探讨热休克蛋70(heat shock protein 70,Hsp70)抑制剂PES(2-phenylethynesulfonamide)对人胃癌BGC细胞活性、细胞迁移以及核因子-κB(nuclear factor-κB,NF-κB)信号通路的影响。方法 :采用Hsp70抑制剂PES处理人胃癌BGC细胞后,应用CCK-8(cell counting kit-8)法检测BGC细胞的活性,细胞划痕实验检测BGC细胞的迁移能力,蛋白质印迹法检测NF-κB信号通路成员及其下游血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达,免疫荧光法观察BGC细胞质中p65的表达。结果 :与未经PES处理的BGC细胞(对照组)比较,5、10和20μmol/L的PES处理BGC细胞72 h后,BGC细胞活性和细胞迁移率被明显抑制(P值均<0.05)。20μmol/L的PES处理后,BGC细胞NF-κB信号通路中磷酸化p65(phospho-p65,p-p65)、p65、NF-κB抑制蛋白激酶α(inhibitory protein of NF-κB kinaseα,IKKα)、p-IKKα、p-IKKβ、IKKβ、p-Akt和Akt以及下游VEGF蛋白的表达水平均明显低于对照组(P值均<0.05),且细胞质中p65蛋白的荧光明显减弱。结论 :Hsp70抑制剂PES可抑制胃癌BGC细胞的活性和迁移,下调BGC细胞NF-κB信号通路活性以及下游VEGF蛋白的表达水平,降低BGC细胞质中p65的表达。

Objective： To investigate the effects of heat shock protein 70 （Hsp70）inhibitor 2-phenylethynesulfonamide （PES） on viability, migration, and nuclear factor-κB （NF-κB） signal pathway of human gastric cancer BGC cells. Methods： The viability and migration of human gastric cancer BGC cells after treatment with PES were detected by cell counting kit-8 （CCK-8） and wound healing experiments, respectively. The activity of NF-κB signal pathway and the expression of vascular endothelial growth factor （VEGF） protein of BGC cells after treatment with PES were measured by Western blotting. The expression of p65 in cytoplasm of BGC cells after treatment with PES was determined by immunofluorescence. Results： As compared with BGC cells without treatment with PES （as a control）, the viability and migration ability of BGC cells after treatment with 5, 10 and 20 μmol/L PES for 72 h were significantly inhibited （both P 〈 0.05）. The expression levels of p65, phospho-p65 （p-p65）, inhibitory protein of NF-κB kinase a （IKKa）, p-IKKa, p-IKKβ, IKKβ, p-Akt and Akt in NF-κB signal pathway and VECF protein of BGC cells after treatment with 20μmol/L PES were lower than those of the control cells （all P 〈 0.05）. The fluorescence expression of p65 in cytoplasm of BGC cells after treatment with 20 μmol/L PES was obviously decreased. Conclusion： Hsp70 inhibitor PES can inhibit the proliferation and migration of BGC cells down-regulate the activity of NF-κB signal pathway and the expression level of VEGF protein and decrease the expression of p65 in cytoplasm of BGC cells.