lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌细胞增殖和侵袭的机制研究

The mechanism of lncRNA TSIX inhibiting proliferation and invasion of hepatocellular carcinoma cell by targeting miR-548-a-3p

目的 探讨lncRNA TSIX通过靶向miR-548-a-3p抑制肝细胞癌(HCC)细胞增殖和侵袭的机制。方法 设肝癌Huh7细胞组、lncRNA TSIX mimics组、lncRNA TSIX inhibitor组;各组细胞培养48 h后,采用MTT法测定细胞活力,结晶紫染色测定单克隆形成数目,流式细胞仪测定细胞凋亡水平,transwell腔室测定细胞侵袭水平,RT-PCR法测定细胞lncRNA TSIX、miR-548-a-3p水平。结果 lncRNA TSIX mimics组OD值、存活率、单克隆形成数目、穿膜数低于Huh7细胞组(t=8.725、5.437、120.834、64.321,P<0.05),lncRNA TSIX inhibitor组OD值、存活率、单克隆形成数目、穿膜数高于Huh7细胞组和lncRNA TSIX mimics组(t=4.124、3.784、10.462、7.965、37.192、174.632、30.703、104.562,P<0.05)。lncRNA TSIX mimics组细胞凋亡率、lncRNA TSIX高于Huh7细胞组[(18.52±2.45)vs(15.63±2.13),(4.47±0.22)vs(2.89±0.21)](t=10.531、12.372,P<0.05),lncRNA TSIX inhibitor组细胞凋亡率、lncRNA TSIX低于Huh7细胞组和lncRNA TSIX mimics组[(8.14±2.45)vs(15.63±2.13)、(18.52±2.45),(1.58±0.25)vs(2.89±0.21)、(4.47±0.22)](t=37.385、48.021、30.703、104.562,P<0.05)。lncRNA TSIX mimics组miR-548-a-3p低于Huh7细胞组[(2.39±0.19)vs(4.87±0.20)](t=11.265,P<0.05),lncRNA TSIX inhibitor组miR-548-a-3p高于Huh7细胞组和lncRNA TSIX mimics组[(6.32±0.19)vs(4.87±0.20)、(2.39±0.19)](t=6.612、10.058,P<0.05)。结论 lncRNA TSIX明显抑制HCC细胞增殖、侵袭,促进HCC细胞凋亡;其机制可能与lncRNA TSIX负调节miR-548-a-3p有关。

Objective To investigate the mechanism of lncRNA TSIX inhibiting proliferation and invasion of hepatocellular carcinoma cell by targeting miR-548-a-3p.Methods The liver cancer cell line Huh7 group,lncRNA TSIX mimics group,and lncRNA TSIX inhibitor group were set up.After 48 hours of cell culturing in each group,the cell viability was measured by MTT method,and the number of single clones formed by crystal violet staining was measured,and the apoptosis level was measured by flow cytometry.The cell invasion level was measured by transwell chamber,and the lncRNA TSIX and miR-548-a-3p levels were measured by RT-PCR method.Results The OD value,survival rate,number of colonies formed,and number of membrane penetration in the lncRNA TSIX mimics group were lower than those in the Huh7 group(t=8.725,5.437,120.834,64.321,P<0.05),and the OD value,survival rate,number of colonies formed,number of membrane penetration in the lncRNA TSIX inhibitor group were higher than those in the Huh7 group and lncRNA TSIX inhibitor group(t=4.124,3.784,10.462,7.965,37.192,174.632,30.703,104.562,P<0.05).The apoptotic rate and lncRNA TSIX in the lncRNA TSIX mimics group was higher than those in the Huh7 group[(18.52±2.45)vs(15.63±2.13),(4.47±0.22)vs(2.89±0.21)](t=10.531,12.372,P<0.05),and the apoptotic rate and lncRNA TSIX in the lncRNA TSIX inhibitor group was lower than those in the Huh7 group and the lncRNA TSIX inhibitor group[(8.14±2.45)vs(15.63±2.13),(18.52±2.45);(1.58±0.25)vs(2.89±0.21),(4.47±0.22)](t=37.385,48.021,30.703,104.562,P<0.05).The miR-548-a-3p in lncRNA TSIX mimics group was lower than that in Huh7 cell group[(2.39±0.19)vs(4.87±0.20)](t=11.265,P<0.05),and miR-548-a-3p in lncRNA TSIX inhibitor group was higher than that in Huh7 group and lncRNA TSIX inhibitor group[(6.32±0.19)vs(4.87±0.20),(2.39±0.19)](t=6.612,10.058,P<0.05).Conclusion LncRNA Tsix significantly inhibited the proliferation and invasion of HCC cells and promoted the apoptosis of HCC cells.The mechanism may be related to the negative regulation of miR-548-a-3p by lncRNA Tsix.