摘要目的 探讨长链非编码RNA胃癌高表达转录因子1(GHET1)在肝细胞癌(HCC)中的表达及其与患者预后、肝癌细胞增殖、迁移和侵袭的关系.方法 收集2016年3月至5月中国人民解放军福州总医院手术的20例HCC患者的肝癌和癌旁组织.回顾性收集2012年6月至2013年12月182例HCC患者肝癌组织和临床资料,根据GHET1表达中位值分为GHET1高表达组和低表达组,各91例.Huh7、HepG2细胞分为:空白对照组(Con)用无血清培养基,siRNA-GHET1组转染siRNA-GHET1,阴性对照组(siRNA-NC)转染阴性对照序列.采用实时荧光定量聚合酶链反应检测GHET1表达,CCK-8、Transwell实验和Western印迹分析GHET1对HCC细胞的影响.结果 与癌旁组织比较,肝癌组织中GHET1的mRNA相对表达明显升高;与L02细胞比较,Huh7以及HepG2细胞GHET1的mRNA表达升高,差异均有统计学意义(P＜0.05).GHET1高表达组肿瘤＞5 cm、有血管侵犯、AFP＞400μg/L、Edmonson Ⅰ级、肿瘤无包膜比例较低表达组升高,差异有统计学意义(P＜0.05).生存分析显示GHET1高表达的HCC患者比低表达的患者预后差.多因素Cox回归分析显示,GHET1高表达、有血管侵犯(HR=2.067,95％CI:1.350～3.162)、无肿瘤包膜是HCC患者复发的独立预测因素.Huh7和HepG2细胞转染后,与Con和siRNA-NC组比较,siRNA-GHET1组增殖明显下降;与siRNA-NC组比较,siRNA-GHET1组迁移与侵袭能力下降,E-钙黏蛋白表达升高,纤连蛋白和波形蛋白表达降低,差异有统计学意义(P＜O.05).结论 GHET1在肝癌组织和细胞中表达升高,增加肝癌细胞的增殖、迁移和侵袭能力,为HCC患者预后的独立预测因素,是临床治疗的潜在靶点.

abstractsObjective To investigate the expression of long-chain non-coding RNA gastric cancer high expression transcription factor 1 (GHET1) in hepatocellular carcinoma (HCC) and the correlation with prognosis,cell proliferation,migration and invasion.Methods 20 HCC patients who underwent surgery from Fuzhou General Hospital of Chinese People's Liberation Army from March to May 2016 were included.The HCC tissue and adjacent normal tissue of 182 patients from June 2012 to December 2013 were retrospectively collected.According to the median value of GHET1 expression,it was divided into GHET1 high expression group and low expression group,91 cases each.Huh7 and HepG2 cells were divided into:blank control group (Con) with serum-free medium,siRNA-GHET1 group transfected with siRNA-GHET1,and negative control group (siRNA-NC) transfected with negative control sequence.The expression of GHET1 was detected by real-time fluorescence quantitative polymerase chain reaction,and the effect of GHET1 on HCC cells was analyzed by CCK-8,Transwell assay and Western blot.Results Compared with adjacent normal tissue,the relative expression of GHET1 mRNA in HCC tissues was significantly increased.Compared with LO2 cells,the mRNA expression of GHET1 in Huh7 and HepG2 cells was higher (P＜0.05).The GHET1 high expression group had tumor＞5 cm,vascular invasion,AFP＞400 μg/L,Edmonson grade Ⅰ,and the tumor-free ratio was lower in the expression group (P＜0.05).Survival analysis showed that HCC patients with high GHET1 expression had a poorer prognosis than patients with low expression.Multivariate Cox regression analysis showed that high expressed GHET1,vascular invasion (HR=2.067,95％ CI:1.350 to 3.162),and without tumor capsule are independent predictors of recurrence in HCC patients.After transfection with Huh7 and HepG2 cells,the proliferation of siRNA-GHET1 group was significantly decreased comparing with Con and siRNA-NC groups.Compared with siRNA-NC group,the migration and invasion ability of siRNA-GHET1 group decreased,and E-cadherin expression increased.The expression of fibronectin and vimentin decreased,and the difference was statistically significant (P＜0.05).Conclusions The expression of GHET1 in HCC tissue is higher comparing with normal tissue,which increases the proliferation,migration and invasion of hepatoma cells.It is an independent predictor of prognosis in HCC patients and a potential target for clinical treatment.