过表达的长链非编码RNA CDKN2B-AS1是肝癌的独立预后因素，并促进其增殖。

Overexpressed lncRNA CDKN2B-AS1 is an independent prognostic factor for liver cancer and promotes its proliferation.

作者信息

Zhuang Huijie, Cao Gang, Kou Changhua, Li Dechun

机构信息

Department of General Surgery, the First Affiliated Hospital of Soochow University, Suzhou, China.

出版信息

J BUON. 2019 Jul-Aug;24(4):1441-1448.

PMID:31646789

Abstract

PURPOSE

To study the expression of long non-coding RNA (lncRNA) CDKN2B-AS1 in the liver tissue of patients with liver cancer and its effect on the proliferation of liver cancer cell line QGY-7703.

METHODS

The expression of differentially expressed genes in liver cancer tissues and normal liver tissues was analyzed by bioinformatics. The expressions of differentially expressed genes in clinical samples were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) to analyze whether there is a significant difference of CDKN2B-AS1 expression in clinical features of patients with liver cancer. The clinical data were analyzed to find out the correlation between the expressions of differentially expressed genes and the overall survival, tumor size and TNM staging. The Gene Set Enrichment Analysis (GSEA) enrichment analysis was used to predict the function of CDKN2B-AS1. The cell proliferation was measured by cell counting kit-8 (CCK-8) assay. Cell cycle was measured by flow cytometry. Expressions of proteins were detected by Western blotting.

RESULTS

The expression of CDKN2B-AS1 in liver cancer was significantly higher compared to normal tissues. The results of qRT-PCR were consistent with the results of the Cancer Genome Atlas (TGCA). The data of CDKN2B-AS1 showed that the expression of CDKN2B-AS1 was associated with the total survival, tumor size and TNM staging. GSEA results showed that genes are enriched in cell cycle sets and others. Compared with the control, the proliferation of QGY-7703 cells overexpressing CDKN2B-AS1 was significantly increased (p<0.001). Western blotting results showed that the expressions of CDK2 and CDK4 were up-regulated and the expression of P16 was decreased after CDKN2B-AS1 was overexpressed.

CONCLUSIONS

LncRNA CDKN2B-AS1 was highly expressed in liver cancer, and its expression was positively correlated with the overall survival, tumor size and TNM stage. LncRNA CDKN2B-AS1 promoted the proliferation and expression of liver cancer cells.

摘要

目的

研究长链非编码RNA（lncRNA）CDKN2B-AS1在肝癌患者肝组织中的表达及其对肝癌细胞系QGY-7703增殖的影响。

方法

通过生物信息学分析肝癌组织和正常肝组织中差异表达基因的表达情况。采用定量实时聚合酶链反应（qRT-PCR）分析临床样本中差异表达基因的表达，以分析CDKN2B-AS1在肝癌患者临床特征中的表达是否存在显著差异。分析临床数据以找出差异表达基因的表达与总生存期、肿瘤大小和TNM分期之间的相关性。使用基因集富集分析（GSEA）富集分析来预测CDKN2B-AS1的功能。通过细胞计数试剂盒-8（CCK-8）法检测细胞增殖。通过流式细胞术检测细胞周期。通过蛋白质印迹法检测蛋白质表达。

结果

与正常组织相比，CDKN2B-AS1在肝癌中的表达显著更高。qRT-PCR结果与癌症基因组图谱（TGCA）的结果一致。CDKN2B-AS1的数据表明，CDKN2B-AS1的表达与总生存期、肿瘤大小和TNM分期相关。GSEA结果表明基因在细胞周期集等中富集。与对照相比，过表达CDKN2B-AS1的QGY-7703细胞的增殖显著增加（p<0.001）。蛋白质印迹结果表明，过表达CDKN2B-AS1后，CDK2和CDK4的表达上调，P16的表达降低。