lncRNA PCAT19靶向miR-143-3p通过信号通路PI3K/Akt对甲状腺癌细胞增殖和凋亡的影响及机制

目的探讨lncRNA PCAT19对甲状腺癌细胞增殖和凋亡的影响及作用机制。方法实验分为miR-con组、miR-143-3p组、si-con组、si-PCAT19组、pcDNA组、pcDNA-PCAT19组、si-PCAT19+anti-miR-con组、si-PCAT19+anti-miR-143-3p组、miR-con+WT-PCAT19组、miR-con+MUT-PCAT19组、miR-143-3p+WT-PCAT19组、miR-143-3p+MUT-PCAT19组。qRT-PCR检测miR-143-3p和PCAT19的表达水平;Western印迹检测蛋白表达;MTT法检测细胞存活率;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测PCAT19和miR-143-3p的靶向关系。结果相较于正常甲状腺细胞HT-ori3,甲状腺癌细胞BCPAP、TPC-1、SW1736中PCAT19的表达水平显著升高,miR-143-3p的表达水平显著降低(P<0.05)。敲低PCAT19和高表达miR-143-3p抑制甲状腺癌细胞BCPAP增殖,促进细胞凋亡;促进裂解的半胱氨酸天冬氨酸蛋白酶(Cleaved-caspase)-3蛋白的表达,抑制细胞周期蛋白(Cyclin)D1的表达。PCAT19靶向负调控miR-143-3p,miR-143-3p低表达可以部分逆转PCAT19低表达对BCPAP细胞增殖抑制和凋亡促进的作用。敲低PCAT19抑制p-AKT和磷脂酰肌醇3-激酶的p1102催化亚基(PI3Kp110α)的表达,miR-143-3p低表达可逆转PCAT19低表达对p-AKT和PI3Kp110α的抑制作用。结论lncRNA PCAT19可抑制甲状腺癌细胞增殖,促进其凋亡,其机制可能与miR-143-3p及PI3K/AKT信号通路有关。

Effect of lncRNA PCAT19 targeting miR-143-3p on proliferation and apoptosis of thyroid cancer cells via signaling pathway PI3K/Akt

Objective To investigate the effect and mechanism of lncRNA PCAT19 on proliferation and apoptosis of thyroid cancer cells.Methods The miR-con group(transfected with miR-con),miR-143-3p group(transfected with miR-143-3p),si-con group(transfected with si-con),si-PCAT19 group(transfected with si-PCAT19),pcDNA group(transfected with pcDNA),pcDNA-PCAT19 group(transfected with pcDNA-PCAT19),si-PCAT19+anti-miR-con group(co-transfected with si-PCAT19 and anti-miR-con),si-PCAT19+anti-miR-143-3p group(co-transfected with si-PCAT19 and anti-miR-143-3p),miR-con+WT-PCAT19 group(co-transfected with miR-con and WT-PCAT19),miR-con+MUT-PCAT19 group(co-transfected with miR-con and MUT-PCAT19),miR-143-3p+WT-PCAT19 group(co-transfected with miR-143-3p and WT-PCAT19),miR-143-3p+MUT-PCAT19 group(co-transfected with miR-143-3p and MUT-PCAT19)were transfected into thyroid cancer cell BCPAP by liposome method.The expressions of miR-143-3p and PCAT19 were detected by qRT-PCR.Western blot was used to detect protein expression.MTT assay was used to detect cell viability.Flow cytometry was used to detect apoptosis.Dual luciferase reporter assay was used to detect fluorescence activity.Results Compared with normal thyroid cell HT-ori3,the expression of PCAT19 in thyroid cancer cells BCPAP,TPC-1 and SW1736 were significantly increased,and the expression of miR-143-3p was significantly decreased(P<0.05).Knockdown of PCAT19 and high expression of miR-143-3p inhibited the proliferation of thyroid cancer cell BCPAP,promoted apoptosis,promoted the expression of Cleaved-caspase-3 protein,and inhibited the expression of Cyclin D1 protein.PCAT19 targeted negatively regulation of miR-143-3p,and low expression of miR-143-3p partially reversed the effect of low expression of PCAT19 on proliferation inhibition and apoptosis of BCPAP cells.Knockdown of PCAT19 inhibited the expression of p-Akt and PI3Kp110α,and low expression of miR-143-3p reversed the inhibitory effect of low expression of PCAT19 on p-Akt and PI3Kp110α.Conclusions LncRNA PCAT19 could inhibit the proliferation of thyroid cancer cells and promote its apoptosis.The mechanism might be related to miR-143-3p and PI3K/AKT signaling pathways,which will provide new targets for the prevention and treatment of thyroid cancer.