Kong Demao, Wang Xia, Wang Xudong, Wang Zixuan, Wang Fa
Interventional Department, Qingdao Municipal Hospital, Qingdao, China.
J BUON. 2021 Mar-Apr;26(2):409-417.
CDKN2C exerts critical functions during the progression of hepatocellular carcinoma (HCC). Its dysfunction is closely linked to poor prognosis of HCC. This study aimed to uncover the underlying mechanism of CDKN2C in affecting the prognosis of HCC.
Potential miRNAs that could regulate CDKN2c were predicted by bioinformatics, and their differential levels in HCC and normal liver tissues were detected. CDKN2C level in Huh7 and Hep3B cells influenced by the two candidate microRNAs, miRNA-22-3p and miRNA-182-5p, were examined. Correlation between miRNA-22-3p and CDKN2C in HCC was analyzed on LinkedOmics, and further confirmed by Pearson correlation test and dual-luciferase reporter gene assay. Thereafter, the prognostic potential of miRNA-22-3p in HCC was evaluated by Kaplan-Meier method. Furthermore, the regulatory effects of miRNA-22-3p/CDKN2C axis on proliferative ability and cell cycle progression of HCC were assessed.
There were five miRNAs predicted to bind to CDKN2C and among them, miRNA-22-3p and miRNA-182-5p were markedly downregulated in LIHC tissues. In Huh7 and Hep3B cells, miRNA-22-3p negatively regulated CDKN2C level, while transfection of miRNA-182-5p mimic or inhibitor did not influence CDKN2C expression. MiRNA-22-3p was closely linked to poor prognosis of HCC patients. Subsequently, dual-luciferase reporter gene assay verified the binding between miRNA-22-3p and CDKN2C.
Knockdown of miRNA-22-3p suppressed proliferative ability and arrested cell cycle progression, which were reversed by overexpression of CDKN2C. MiRNA-22-3p suppresses proliferative ability and arrests cell cycle progression in HCC through targeting CDKN2C.
CDKN2C在肝细胞癌（HCC）进展过程中发挥关键作用。其功能障碍与HCC的不良预后密切相关。本研究旨在揭示CDKN2C影响HCC预后的潜在机制。
通过生物信息学预测可能调控CDKN2c的潜在miRNA，并检测其在HCC组织和正常肝组织中的差异水平。检测了受两种候选微小RNA（miRNA-22-3p和miRNA-182-5p）影响的Huh7和Hep3B细胞中CDKN2C的水平。在LinkedOmics上分析了HCC中miRNA-22-3p与CDKN2C之间的相关性，并通过Pearson相关性检验和双荧光素酶报告基因测定进一步证实。此后，采用Kaplan-Meier法评估miRNA-22-3p在HCC中的预后潜力。此外，评估了miRNA-22-3p/CDKN2C轴对HCC增殖能力和细胞周期进程的调控作用。
预测有5种miRNA与CDKN2C结合，其中miRNA-22-3p和miRNA-182-5p在LIHC组织中显著下调。在Huh7和Hep3B细胞中，miRNA-22-3p负调控CDKN2C水平，而转染miRNA-182-5p模拟物或抑制剂不影响CDKN2C表达。MiRNA-22-3p与HCC患者的不良预后密切相关。随后，双荧光素酶报告基因测定验证了miRNA-22-3p与CDKN2C之间的结合。
敲低miRNA-22-3p可抑制增殖能力并使细胞周期进程停滞，而CDKN2C的过表达可逆转这一现象。MiRNA-22-3p通过靶向CDKN2C抑制HCC的增殖能力并使细胞周期进程停滞。
