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  • 李墨林,陈航,黄平.环状RNA circSP3(hsa-circ-0002642)促进肝细胞癌细胞增殖及迁移[J].第二军医大学学报,2019,40(12):1330-1336    [点击复制]
  • LI Mo-lin,CHEN Hang,HUANG Ping.Circular RNA circSP3 (hsa-circ-0002642) promotes proliferation and migration of hepatocellular carcinoma cells[J].Acad J Sec Mil Med Univ,2019,40(12):1330-1336   [点击复制]
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环状RNA circSP3(hsa-circ-0002642)促进肝细胞癌细胞增殖及迁移
李墨林1,陈航2,黄平1*
0
(1. 重庆医科大学附属第一医院肝胆外科, 重庆 400016;
2. 重庆市铜梁区人民医院肿瘤血液内科, 重庆 402560
*通信作者)
摘要:
目的 探讨环状RNA circSP3(hsa-circ-0002642)在肝细胞癌(HCC)中的表达及其对HCC细胞增殖及迁移的影响。方法 收集重庆医科大学附属第一医院肝胆外科2017年6月至2018年12月收治的42例HCC患者手术切除的癌组织和癌旁组织标本,采用实时定量PCR检测组织中circSP3的表达,分析癌组织中circSP3表达与患者临床病理学指标的关系。培养人HCC细胞系Hep-3B、Huh7、SMMC-7721、Bel-7402及人正常肝细胞系HL-7702,采用实时定量PCR检测细胞中circSP3的表达。使用circSP3过表达及干扰质粒分别转染Hep-3B和Huh7细胞后,采用细胞计数试剂盒-8(CCK-8)检测细胞增殖情况,Transwell法检测细胞侵袭和迁移能力,蛋白质印迹法检测上皮-间质转化(EMT)相关标志物波形蛋白、E-钙黏蛋白的表达。结果 HCC患者癌组织中circSP3的表达高于癌旁组织(P<0.01),并且circSP3的表达与HCC肿瘤最大径及TNM分期呈正相关(P均<0.05)。circSP3在4种HCC细胞系中的表达均高于正常肝细胞系(P均<0.01)。circSP3过表达可以促进HCC细胞的增殖、侵袭和迁移(P<0.05、P<0.01),而干扰circSP3表达则抑制HCC细胞的增殖、侵袭和迁移(P<0.05、P<0.01)。波形蛋白表达在circSP3过表达的HCC细胞中高于对照细胞(P<0.05),而在干扰circSP3表达的HCC细胞中低于对照细胞(P<0.05)。E-钙黏蛋白表达在circSP3过表达的细胞中低于对照细胞(P<0.01),而在干扰circSP3表达的HCC细胞中高于对照细胞(P<0.01)。结论 circSP3的异常表达与HCC肿瘤大小及TNM分期呈正相关,有助于评价病情及预后。circSP3能促进HCC细胞的增殖,并且可能通过促进EMT进程进而促进HCC细胞的迁移和侵袭。
关键词:  环状RNA  circSP3  肝细胞癌  上皮-间质转化  细胞增殖  迁移  侵袭
DOI:10.16781/j.0258-879x.2019.12.1330
投稿时间:2019-07-27修订日期:2019-11-11
基金项目:
Circular RNA circSP3 (hsa-circ-0002642) promotes proliferation and migration of hepatocellular carcinoma cells
LI Mo-lin1,CHEN Hang2,HUANG Ping1*
(1. Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;
2. Department of Oncology and Hematology, The People's Hospital of Tongliang District, Chongqing 402560, China
*Corresponding author)
Abstract:
Objective To investigate the expression of circular RNA circSP3 (hsa-circ-0002642) in hepatocellular carcinoma (HCC) and its effect on proliferation and migration of HCC cells. Methods The tumor tissue and the adjacent paratumor tissue samples were collected from 42 HCC patients who underwent surgery from Jun. 2017 to Dec. 2018 in Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University. The expression of circSP3 in the samples was detected by real-time quantitative PCR, and the relationship between circSP3 expression in the tumor tissues and clinicopathological parameters of the patients was analyzed. Human HCC cell lines (Hep-3B, Huh7, SMMC-7721 and Bel-7402) and human normal liver cell line (HL-7702) culturion, and the expression of circSP3 was detected. After circSP3 overexpression and interference plasmids transfection into Hep-3B and Huh7 cells, the cell proliferation was detected by cell counting kit-8 (CCK-8) method, the invasion and migration abilities were detected by Transwell assay, and expression levels of epithelial-mesenchymal transformation (EMT)-associated markers (vimentin and E-cadherin) were determined by Western blotting. Results The expression of circSP3 of tumor tissues was significantly higher than that of the paratumor tissues in the HCC patients (P<0.01), and the expression of circSP3 was positively correlated with the tumor maximum diameter and clinical TNM stage (both P<0.05). The expression levels of circSP3 in the 4 HCC cell lines were significantly higher than that in the normal liver cell lines (all P<0.01). Overexpression of circSP3 could significantly promote proliferation, invasion and migration of HCC cells (P<0.05, P<0.01), while interference circSP3 expression could significantly inhibit the proliferation, invasion and migration (P<0.05, P<0.01). The expression of vimentin was significantly higher in circSP3-overexpressed cells than that in control cells (P<0.05), while it was significantly lower in circSP3-interfered cells than that in control cells (P<0.05). The expression of E-cadherin was significantly lower in circSP3-overexpressed cells than that in control cells (P<0.01), while it was significantly higher in circSP3-interfered cells than that in control cells (P<0.01). Conclusion The abnormal expression of circSP3 is positively correlated with tumor size and TNM stage of HCC, and determining its expression is helpful to evaluate the severity and prognosis of HCC. CircSP3 can promote the proliferation of HCC cells, and may promote the migration and invasion by promoting the EMT process.
Key words:  circular RNAs  circSP3  hepatocellular carcinoma  epithelial-mesenchymal transformation  cell proliferation  migration  invasion