YAP1基因沉默诱导甲状腺癌细胞凋亡及逆转免疫抑制的机制研究

Mechanism of YAP1 Gene Silencing-Induced Apoptosis and Immunosuppression Reversal in Thyroid Cancer Cells

目的探讨沉默K4P1基因表达对甲状腺癌细胞凋亡及免疫抑制因子表达的影响。方法将实验分为空白组(未进行处理的K1细胞)、NC组(K1细胞转染阴性对照siRNA)和si-YAP1组(K1细胞转染YAP1特异性siRNA)。MTT法检测siRNA转染4d细胞活力;流式细胞术及Western印迹分别检测si-YAP1转染K1细胞48h的细胞凋亡率及YAP1、VEGF、TGF-β1、AKT、p-AKT和Bax蛋白表达。结果si-YAP1的转染可明显降低K1细胞YAP1蛋白表达,与空白组比较有显著性差异(P<0.05)。si-YAP1组和空白组比较,细胞活力明显降低,凋亡率明显升高,VEGF、TGF-β1和p-AKT蛋白表达明显降低,Bax蛋白表达明显升高(P<0.05)。结论沉默表达可诱导甲状腺癌细胞凋亡,且逆转免疫抑制,机制可能与PI3K/AKT信号的下调有关。

Objective To investigate the effect of YAP1 gene silencing on apoptosis and expression of immunosuppressive factors in thyroid carcinoma cells. Methods Cells was divided into blank group ( untreated KI cells), NC group ( negative control siRNA was transfected into KI cells) and si-YAP1 group ( YAP1 specific siRNA was transfected into KI cells). MTT assay was used to detect the viability of cells siRNA was transfected into KI cells for 4 days. Flow cytometry and Western blotting were used to detect the apoptosis rate and the expression of YAP1 , VEGF, TGF-β1 , AKT, p-AKT and Bax proteins in KI cells siRNA was transfected with si-YAP1 for 48 hours. Results The expression of YAP1 protein in KI cells was significantly decreased after si- YAP1 transfection compared with the blank group ( P < 0. 05 ). Compared with the blank group, the cell viability was significantly decreased, the apoptosis rate was significantly increased, the expressions of VEGF, TGF-β1 and p-AKT protein were significantly decreased, and the expression of Bax protein was significantly increased in YAP1 knockdown group ( P < 0. 05 ). Conclusion Silencing YAP1 expression can induce apoptosis of thyroid carcinoma cells and reverse immunosuppression , which may be related to the down-regulation of PI3K/AKT signal.