长链非编码RNA linc00261在肝细胞癌中的表达及意义

Expression of long noncoding RNA linc00261 in hepatocellular carcinoma and its association with postoperative outcomes

目的研究长链非编码RNA linc00261在肝细胞癌(HCC)中的表达、功能及其临床意义。方法采用实时荧光定量PCR(qRT-PCR)法检测linc00261在74例肝癌、癌旁肝组织中的表达;采用卡方检验分析其表达量与临床病理指标的相关性;应用COX回归模型分析其表达水平与肝癌患者术后预后的关系;qRT-PCR检测linc00261在5株肝癌细胞株中的表达;在肝癌细胞株MHCC-LM3和SNU-449中采用siRNA(si-linc00261-1,si-linc00261-2,si-NC和空白对照组)和Lipofectamine3000进行转染敲低linc00261,活细胞计数(CCK8)、Transwell实验分别研究linc00261对肝癌细胞增殖,侵袭迁移能力的影响。结果 HCC组织中linc00261的表达水平与肝癌患者术前血清AFP(P=0.032)、肿瘤大小(P=0.007)、有无微血管癌栓(P=0.01)、TNM分期(P=0.02)显著相关;HCC组织中linc00261的低表达与患者的不良预后相关,其术后无瘤生存期明显缩短(P<0.05),是影响HCC患者术后无瘤生存期的一个独立危险因素;下调linc00261的表达可促进肝癌细胞株MHCC-LM3(P<0.01)和SNU-449(P<0.001)的迁移和侵袭能力,对细胞增殖无明显影响(P>0.05)。结论 Linc00261有望成为肝癌切除术后的一种新的预后指标。

Objective To investigate the expression of long noncoding RNA linc00261 in hepatocellular carcinoma（HCC） and its correlation with the clinicopathological features and postoperative outcomes of the patients. Methods Real-time fluorescence quantitative PCR（qRT-PCR） was used to detect the expression of linc00261 in surgical specimens of HCC and adjacent tissues from 74 patients. The correlation of the expression level of linc00261 in HCC with the clinicopathological parameters of the patients was analyzed using Chi-square test. The Cox＇s proportional hazards regression model was used to assess the value of linc00261 in predicting the prognosis of HCC patients after operation. The expression of linc00261 was also examined in 5 HCC cell lines using qRT-PCR. The HCC cell lines MHCC-LM3 and SNU-449 were transfected with small interfering RNAs targeting linc00261 for linc00261 knockdown, and the changes in the cell proliferation, migration and invasion abilities were observed using CCK-8 assay and Transwell assay. Results The expression level of linc00261 in HCC was significantly correlated with AFP（P=0.032）, tumor size（P=0.007）, microscopic vascular invasion（MVI; P=0.01）, and TNM stage（P=002）. The patients with lowered expressions of linc00261 in HCC tissues had a significantly shortened tumor-free survival time（P〈0.05）,and a lowered expression of linc00261 was identified as an independent risk factor affecting postoperative recurrence-free survival time of the patients（P〈0.05）. In HCC cell lines MHCC-LM3 and SNU-449 cells, linc00261 knockdown obviously promoted the cell migration and invasion（P〈0.01） but did not significantly affect cell proliferation（P〈0.05）. Conclusion Linc00261 may serve as a new prognostic biomarker for predicting the postoperative outcomes of the patients with HCC.