摘要目的:探讨甲基转移酶样蛋白14(METTL14)介导长链非编码RNA EIF3J反义RNA1(lnc EIF3J-AS1)异常表达对胆管癌细胞迁移和侵袭能力的影响及其作用机制。方法:收集2017年9月至2018年12月间海军军医大学第一附属医院行手术切除并经病理学确诊的10例胆管癌组织及其相应癌旁正常组织，采用荧光定量PCR法检测胆管癌组织METTL14 mRNA和lnc EIF3J-AS1的表达，采用蛋白质免疫印迹法检测METTL14的蛋白表达。选取胆管癌细胞株HUCCT1和RBE，分为对照组和METTL14或lnc EIF3J-AS1敲低组，对照组分别转染相应的阴性对照的慢病毒，METTL14或lnc EIF3J-AS1敲低组分别转染干扰METTL14或lnc EIF3J-AS1表达的慢病毒。采用Transwell实验检测各组细胞迁移和侵袭能力，采用蛋白质免疫印迹法检测各组细胞表皮生长因子受体(EGFR)和AKT蛋白表达。结果:胆管癌组织METTL14 mRNA和lnc EIF3J-AS1表达量均显著高于癌旁正常组织(0.075±0.012比0.031±0.006，0.140±0.032比0.064±0.012)，且METTL14 mRNA与lnc EIF3J-AS1表达水平呈正相关( r＝0.883， P＝0.0007)。胆管癌组织METTL14蛋白表达量高于癌旁正常组织(0.354±0.131比0.187±0.183)。与对照组相比，胆管癌细胞株HUCCT1和RBE METTL14敲减组lnc EIF3J-AS1表达水平明显下降(0.217±0.020比1.000±0.052，0.149±0.066比1.000±0.045)。HUCCT1细胞和RBE细胞lnc EIF3J-AS1敲减组迁移和侵袭能力明显下降(5.00±0.58比23.33±0.33，20.33±0.67比70.67±0.33；12.00±0.58比25.00±2.52，22.33±0.89比43.67±0.33)，且EGFR与p-AKT/AKT蛋白表达水平也明显下降(0.109±0.015比1.000±0.018，0.226±0.036比1.000±0.051；0.118±0.052比1.000±0.069，0.132±0.098比1.000±0.023)。以上差异均有统计学意义( P值均<0.05)。 结论:METTL14介导胆管癌中lnc EIF3J-AS1异常表达可促进肿瘤细胞迁移和侵袭。

abstractsObjective:To investigate the effects of methyltransferase-like protein 14 (METTL14)-mediated long-chain non-coding RNA EIF3J antisense RNA1 (Inc EIF3J-AS1) on the migration and invasion of cholangiocarcinoma cells and its mechanism.Methods:From September 2017 to December 2018, 10 pairs of cholangiocarcinoma and adjacent normal tissues were collected from the First Affiliated Hospital of Naval Medical University, which were surgically resected and pathologically confirmed. The expression of METTL14 mRNA and Inc EIF3J-AS1 in cholangiocarcinoma tissues was detected by fluorescence quantitative PCR, and the protein expression of METTL14 was detected by Western blotting. Cholangiocarcinoma cell lines HUCCTI and RBE were divided into control group and METTL14 or Inc EIF3J-AS1 knockdown group. The corresponding normal lentivirus was transfected in the control group, and METTL14 or Inc EIF3J-AS1 knockdown group was transfected with lentivirus that interfered with the expression of METTL14 or Inc EIF3J-AS1, respectively. Transwell assay was used to detect the ability of cell migration and invasion, and Western blotting was used to detect the expression of epidermal growth factor receptor (EGFR) and AKT protein.Results:The expressions of METTL14 mRNA and lnc EIF3J-AS1 in cholangiocarcinoma tissues were significantly higher than those in adjacent normal tissues (0.075±0.012 vs 0.031±0.006, 0.140±0.032 vs 0.064±0.012), and there was a positive correlation between expression of METTL4 mRNA and expression of lnc EIF3J-AS1 ( r=0.883, P=0.0007). The expression of METTL14 protein in cholangiocarcinoma tissues was higher than that in adjacent normal tissues (0.354±0.131 vs 0.187±0.183). Compared with the control group, the expression of lnc EIF3J-AS1 was significantly lower in METTL14 or Inc EIF3J-AS1 knockdown group (0.217±0.020 vs 1.000±0.052, 0.149±0.066 vs 1.000±0.045). The migration and invasion ability of cell lines HUCCTI and RBE decreased significantly in lnc EIF3J-AS1 knockout group (5.00±0.58 vs 23.33±0.33, 20.33±0.67 vs 70.67±0.33; 12.00±0.58 vs 25.00±2.52, 22.33±0.89 vs 43.67±0.33). The expression of EGFR and p-AKT/AKT protein were also significantly decreased (0.109±0.015 vs 1.000±0.018, 0.226±0.036 vs 1.000±0.051; 0.118±0.052 vs 1.000±0.069, 0.132±0.098 vs 1.000±0.023). The above differences were statistically significant (all P<0.05). Conclusions:Abnormal expression of lnc EIF3J-AS1 in cholangiocarcinoma mediated by METTL14 can promote tumor cell migration and invasion.