下调lncRNA FAL1对甲状腺癌TPC-1细胞功能的影响及其机制

Effects of Down-regulated LncRNA FAL1 on the Function of Thyroid Cancer TPC-1 Cells and Its Mechanism

目的探讨下调lncRNA FAL1对甲状腺癌TPC-1细胞增殖、周期分布、凋亡、侵袭和迁移的影响及其机制。方法将体外培养的TPC-1细胞分为空白对照组、阴性对照组和FAL1干扰组,采用实时荧光定量PCR检测TPC-1细胞中FAL1表达水平,MTT实验检测TPC-1细胞增殖活力,流式细胞仪检测TPC-1细胞周期分布和凋亡,Transwell小室检测TPC-1细胞侵袭和迁移,Western blotting检测TPC-1细胞中AKT磷酸化和CyclinD1、Bcl-2、MMP-9蛋白表达水平。结果与空白对照组比较,阴性对照组TPC-1细胞中各指标差异无统计学意义(P>0.05);与空白对照组或阴性对照组比较,FAL1干扰组TPC-1细胞增殖活力、S期细胞百分比、细胞侵袭能力、迁移能力和AKT磷酸化以及CyclinD1、Bcl-2、MMP-9蛋白的表达水平均明显降低,而G0/G1期细胞百分比和细胞凋亡率均明显升高(P<0.05)。结论下调FAL1表达可抑制甲状腺癌TPC-1细胞增殖、侵袭和迁移并诱导细胞周期阻滞和凋亡,其作用机制可能与抑制AKT信号通路有关。

Objective To investigate the effect of down-regulated lncRNA FAL1 on proliferation,cycle distribution,apoptosis,invasion and migration of thyroid cancer TPC-1 cells and its mechanism.Methods TPC-1 cells cultured in vitro were divided into the blank control group,negative control group and FAL1 interference group.The expression of FAL1 in TPC-1 cells was detected by real-time fluorescence quantitative PCR.The proliferation activity of TPC-1 cells was detected by MTT assay.Cell cycle distribution and apoptosis of TPC-1 cells were detected by cytometry,TPC-1 cell invasion and migration were detected by Transwell chamber,and AKT phosphorylation and CyclinD1,Bcl-2,MMP-9 protein expression levels in TPC-1 cells were detected by western blotting.Results Compared with the blank control group,there was no significant difference in the parameters of the TPC-1 cells in the negative control group(P>0.05).Compared with the blank control group or the negative control group,the proliferation activity of TPC-1 cells,the percentage of S-phase cells,cell invasion ability,migration ability and AKT phosphorylation and the expression levels of CyclinD1,Bcl-2,MMP-9 proteins were significantly reduced in the FAL1 interference group,while the percentage of cells and the apoptosis rate in G0/G1 phase were significantly increased(P<0.05).Conclusion Down-regulating of FAL1 expression can inhibit the proliferation,invasion and migration of thyroid cancer TPC-1 cells,and induce cell cycle arrest and apoptosis.The mechanism could be related to the inhibition of AKT signaling pathway.