miR-128-3p 通过靶向调控 CDC6 抑制肝癌细胞的增殖和转移
MiR-128-3p suppresses tumor proliferation and metastasis via targeting CDC6 in hepatocellular carcinoma cells.
机构信息
Department of Hepatobiliary Surgery, Xinchang People's Hospital, China; School of Medicine, Shaoxing University, China.
Department of Hepatobiliary Surgery, Xinchang People's Hospital, China; Wuhan University, China.
出版信息
Tissue Cell. 2021 Oct;72:101534. doi: 10.1016/j.tice.2021.101534. Epub 2021 Mar 23.
BACKGROUND
MicroRNAs (miRNAs) are known to be involved in the pathogenesis of various cancers. The present study devotes efforts to discover the role of miR-128-3p in hepatocellular carcinoma (HCC).
METHODS
MiR-128-3p and cell division cycle 6 (CDC6) expressions in HCC tissue (n = 50) and adjacent normal tissue (n = 50) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). MTT assay and flow cytometry were applied to measure the viability and cell cycle distribution of HuH7 and HCCLM3 cells, respectively. The potential binding sites of miR-128-3p on CDC6 were predicted with Targetscan 7.2 and confirmed by dual-luciferase reporter assay. Expression analysis of CDC6 and survival analysis in HCC were performed by GEPIA2. Wound healing and Transwell assays were used to detect HCC cell migration and invasion, respectively. Expressions of miR-128-3p and epithelial-mesenchymal transition (EMT)-related proteins (MMP2, MMP9, E-Cadherin, N-Cadherin and Vimentin) were quantified using qRT-PCR and western blot, respectively.
RESULTS
MiR-128-3p mRNA expression was lower in HCC tissue than in adjacent normal tissues. HCC cell viability was suppressed and cell cycle was arrested in G0/S phase by miR-128-3p mimic. CDC6 was targeted by miR-128-3p and had higher expression in HCC tissue. The promotive effects of overexpressed CDC6 on HCC cell viability, migration and invasion were reversed by up-regulating miR-128-3p. And the effects of overexpressed CDC6 on inhibiting E-Cadherin expression yet promoting MMP2, MMP9, N-Cadherin and Vimentin expressions in HCC cells were reversed by up-regulating miR-128-3p.
CONCLUSION
MiR-128-3p may suppress HCC cell proliferation and metastasis via targeting CDC6.
背景
微 RNA（miRNA）已被证实参与多种癌症的发病机制。本研究旨在探究 miR-128-3p 在肝细胞癌（HCC）中的作用。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测 50 例 HCC 组织和 50 例癌旁正常组织中 miR-128-3p 和细胞分裂周期蛋白 6（CDC6）的表达。MTT 法和流式细胞术分别用于检测 HuH7 和 HCCLM3 细胞的活力和细胞周期分布。Targetscan 7.2 预测 miR-128-3p 与 CDC6 的潜在结合位点，并通过双荧光素酶报告基因实验进行验证。利用 GEPIA2 进行 CDC6 表达分析和 HCC 患者生存分析。划痕愈合和 Transwell 实验分别用于检测 HCC 细胞迁移和侵袭。采用 qRT-PCR 和 Western blot 分别定量检测 miR-128-3p 和上皮-间质转化（EMT）相关蛋白（MMP2、MMP9、E-钙黏蛋白、N-钙黏蛋白和波形蛋白）的表达。
结果
与癌旁正常组织相比，miR-128-3p 在 HCC 组织中的表达降低。miR-128-3p 模拟物抑制 HCC 细胞活力，将细胞周期阻滞在 G0/S 期。CDC6 是 miR-128-3p 的靶基因，在 HCC 组织中表达较高。过表达 CDC6 对 HCC 细胞活力、迁移和侵袭的促进作用被上调 miR-128-3p 所逆转。上调 miR-128-3p 还逆转了过表达 CDC6 抑制 HCC 细胞中 E-钙黏蛋白表达、促进 MMP2、MMP9、N-钙黏蛋白和波形蛋白表达的作用。
结论
miR-128-3p 可能通过靶向 CDC6 抑制 HCC 细胞增殖和转移。
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