维替泊芬对鼻息肉上皮细胞Yes相关蛋白的作用

Effect of verteporfin on Yes-associated protein in human nasal polyps epithelial cells

目的研究Ki-67和Yes相关蛋白(YAP)在鼻息肉组织中的表达,探讨维替泊芬对人鼻息肉上皮细胞(hNPEC)增殖的影响。方法实时荧光定量PCR及蛋白免疫印迹法分别检测正常下鼻甲黏膜组织和鼻息肉组织的Ki-67和YAP的mRNA及蛋白水平。体外培养hNPEC,噻唑蓝比色法(MTT)确定维替泊芬的半数抑制浓度(IC_(50)),蛋白免疫印迹法检测维替泊芬IC_(50)处理组中Ki-67、YAP及TEAD1的蛋白表达,免疫荧光染色观察Ki-67、YAP的细胞内表达。结果在鼻息肉组织中, Ki-67和YAP的mRNA和蛋白表达均高于下鼻甲对照组,差异有统计学意义(P均<0.05)。维替泊芬作用hNPEC 24 h后IC_(50)为2.516μmol/L。维替泊芬下调hNPEC中Ki-67及YAP、TEAD1的蛋白表达水平,差异有统计学意义(P均<0.05)。免疫荧光染色显示维替泊芬组中Ki-67阳性细胞数较少,YAP核内荧光强度明显降低。结论维替泊芬可能通过抑制YAP-TEAD复合物来抑制hNPEC的增殖。

Objective To investigate the expression levels of Ki-67 and Yes-associated protein(YAP) in the nasal polyps tissues and evaluate the effect of verteporfin on the proliferation of human nasal polyps epithelial cells(hNPECs). Methods The expression of Ki-67 and YAP at the mRNA and protein levels in normal inferior turbinate mucosa(n=10) and nasal polyps tissues(n=20) were detected by quantitative PCR(qPCR) and Western blot. The hNPECs were cultured in vitro. The half maximal inhibitory concentration(IC50) of verteporfin at 24 h was determined by MTT assay. The expression levels of Ki-67, YAP and TEAD1 proteins in hNPECs treated with IC50 of verteporfin were detected by Western blot. The expression of Ki-67 and YAP in hNPECs was observed by immunofluorescence staining. Results The expression of Ki-67 and YAP at the mRNA and protein levels in the nasal polyps tissues was significantly higher compared with those in the control group(all P<0.05). MTT assay demonstrated that the IC50 of verteporfin was 2.516 μmol/L in hNPECs after 24-h incubation. The expression levels of Ki-67, YAP and TEAD1 proteins in hNPECs were significantly down-regulated by verteporfin(all P<0.05). Immunofluorescence staining revealed that the quantity of Ki-67 positive cells was considerably decreased and the fluorescence intensity of nucleus YAP was significantly reduced in the verteporfin group. Conclusions Verteporfin inhibits the proliferation of hNPECs probably by suppressing the YAP-TEAD complex.