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Hsa_circ_PVT1在肝细胞癌中的表达及意义
作者:朱元鑫 付航玮 林夏 蓝翔 马钰 李光耀 董睿 王宇洲 陈平 
单位:400042  重庆  陆军军医大学大坪医院野战外科研究所肝胆外科(朱元鑫、付航玮、林夏、蓝翔、马钰、李光耀、董睿、王宇洲、陈平) 
关键词:肝细胞癌 环状RNA PVT1 细胞增殖 细胞周期 
分类号:R322.47;R329.28;R394.1
出版年,卷(期):页码:2018,43(3):211-216
摘要:
[摘要目的 探讨人肝细胞癌(HCC)circ-PVT1的表达及其对肝癌细胞增殖的影响,并探讨其临床意义。方法 利用RT-qPCR技术检测46例人肝癌组织/癌旁组织中circ-PVT1的表达水平,并且分析病理指标与其表达水平的关系;体外培养人正常肝细胞系(L02)、人肝癌细胞系(HepG2SMMC-7721MHCC-97HMHCC-97LHCC-LM3),利用RT-qPCR技术检测circ-PVT1的表达水平。利用脂质体转染技术体外转染si-circPVT1,敲低circ-PVT1HepG2SMMC-7721细胞系中的表达量,同时设置阴性对照组,通过CCK-8实验、EdU实验检测干扰circ-PVT1表达对肝癌细胞增殖的影响;采用流式细胞术观察干扰circ-PVT1表达对肝癌细胞周期的影响。结果 circ-PVT1在人肝癌组织中表达量明显高于癌旁组织(P<0.001),且其表达水平与肿瘤大小、TNM分期及分化程度相关;同样,肝癌细胞系HepG2SMMC-7721MHCC-97HMHCC-97LHCC-LM3circ-PVT1表达水平也明显高于正常肝细胞系L02(P<0.05);与阴性对照组比较,circ-PVT1干扰组HepG2SMMC7721的增殖能力明显降低。结论 在人HCC的诊断中,circ-PVT1可能作为一种潜在的生物标志物予以应用,并且可能成为一种新型的增殖因子。
[Abstract]  Objective  To determine the expression and clinical significance of circ-PVT1 in human hepatocellular carcinoma (HCC) and its effect on HCC cell proliferation. Methods The expressions of circ-PVT1 in hepatocellular carcinoma and the matched tumor-adjacent tissues were detected by RT-qPCR and the relationship between pathological indexes and the expression level was analyzed in 46 patients. The expressions of circ-PVT1 in human normal liver cell line (L02) and hepatocellular carcinoma cell lines (HepG2, SMMC-7721, MHCC-97H, MHCC-97L, HCC-LM3) were detected by RT-qPCR and were compared thereafter. With knocking down the expression of circ-PVT1, si-circPVT1 was transfected into HepG2 and SMMC-7721 cells by using lipofectamine technique in vitro, with the si-NC being taken as negative control. After interfering the expression of circ-PVT1, the effect on the proliferation of hepatocellular carcinoma cells was detected by CCK-8 and EDU experiments and flow cytometry was conducted to observe the effect of circ-PVT1 on cell cycle. Results The expression level of circ-PVT1 was significantly higher in HCC tissues than in adjacent tissues (P<0.01), and its high expression level was significantly correlated with tumor size, TNM stage and differentiation degree. Similarly, in human hepatocellular carcinoma cell lines (HepG2, SMMC-7721, MHCC-97H, MHCC-97L, HCC-LM3), the expression level of circ-PVT1 was also higher than that in human normal liver cell line L02 (P<0.05). Compared with the negative control group, silencing of circ-PVT1 resulted in remarkable reduction in cell proliferation of HepG2 and SMMC-7721. Conclusion  circ-PVT1 may act as a potential biomarker for HCC diagnosis and may become a novel proliferation factor.
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