长链非编码RNA 00460表达下调对胆管癌HuCCT1细胞生物学行为的影响

Effect of down-regelation of long noncoding RNA 00460 on biological behavior of cholangiocarcinoma HuCCT1 cells

目的探讨长链非编码RNA 00460(LINC00460)对胆管癌HuCCT1细胞增殖、侵袭和迁移能力的影响。方法分析LINC00460在正常胆管细胞及胆管癌细胞HuCCT1的表达差异。空载病毒及LINC00460KD病毒感染HuCCT1后,应用实时荧光定量PCR检测LINC00460表达。应用CCK-8法、平板克隆、流式细胞仪检测下调LINC00460对HuCCT1增殖、克隆能力及周期的影响;应用Transwell小室实验检测下调LINC00460对HuCCT1侵袭和迁移能力的影响。蛋白印记检测法检测相关蛋白的表达。结果胆管癌细胞HuCCT1中的表达水平明显高于正常胆管细胞(P<0.05), LINC00460KD病毒感染胆管癌细胞株后,LINC00460表达水平量明显减少。与对照组相比,下调LINC00460后,HuCCT1的增殖活力明显降低, G1期比例明显升高(P<0.05)。同时,下调LINC00460胆管癌细胞HuCCT1侵袭迁移能力明显减弱(P<0.05)。CyclinD1、CDK2和vimentin表达减少,E-cadherin表达明显增多。结论 LINC00460在胆管癌细胞HuCCT1中高表达且促进胆管癌细胞HuCCT1的增殖、迁移和侵袭。

Objective To investigate the effect of LINC00460 on proliferation, invasion and migration of cholangiocarcinoma HuCCT1 cells. Methods The expression of LINC00460 in HuCCT1 in human intrahepatic biliary epithelial cells and cholangiocarcinoma cells was analyzed. The expression of LINC00460 was detected by real-time fluorescence PCR after the HuCCT1 cells were infected with LINC00460 KD virus. The effects of down-regulation of LINC00460 on proliferation, clone ability and cycle of HuCCT1 were assessed by CCK-8, clonal assay and flow cytometry. The effect of down-regulation of LINC00460 on the invasion and migration of HuCCT1 was assessed by trans-well chamber assay. The expression of cyclinD1, CDK2, E-cadherin and vimentin were assessed by western blot. Results The expression of LINC00460 in HuCCT1 cells was significantly higher than that in normal cell(P<0.05). After infected with LINC00460 KD, LINC00460 expression level in HuCCT1 cells was down-regualted. Down-regulation of LINC00460 reduced cell proliferation and colony formation. Down-regulation of LINC00460 resulted in G0/G1 cell cycle arrest compared to the controls. Trans-well chamber assay found that down-regulation of LINC00460 significantly inhibited the migration and invasion ability of HuCCT1 cells in experimental group(P<0.05). Western blotting verified that cyclinD1, CDK2 and vimentin was significantly reduced, while E-cadherin was increased in HuCCT1 cell with the down-regulation of LINC00460. Conclusion LINC00460 is highly expressed in cholangiocarcinoma HuCCT1 cells and promotes their proliferation, migration and invasion.