抑制 S 期激酶相关蛋白 2 基因 (SKP2) 可抑制肝细胞癌细胞的增殖和迁移

IF 1.5 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biology Pub Date : 2024-09-11 DOI:10.1134/s0026893324700651
Y. Y. Zhao, Z. X. Gao, S. D. Wei, W. Song
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引用次数: 0

摘要

摘要SKP2基因是某些疾病的独立预后因子,也是一种潜在的癌基因。肝癌发生和发展的分子机制以及SKP2在这一过程中的参与尚不清楚。在此,为了研究SKP2对肝癌细胞增殖、凋亡和迁移的影响,我们利用慢病毒介导的RNA干扰技术,使用特异性SKP2的短发夹RNAs(shRNAs)。结果表明,与 379 例正常肝组织相比,SKP2 在 809 例肝癌组织中的表达明显上调。SKP2 mRNA水平高的患者生存时间比水平低的患者短,SKP2在III期肝细胞癌组织中表达量最高。通过 MTT 试验、流式细胞术、集落形成试验、Transwell 和 Western 印迹分析,评估了沉默 SKP2 对 Huh7 和 HepG2 细胞增殖、凋亡、细胞周期、迁移和凋亡蛋白表达的影响。在稳定转染的Huh7和HepG2细胞中,SKP2的表达明显降低,敲除率分别为95.7%和85.8%。转染癌细胞的活力、增殖和迁移均有所降低。这些细胞的凋亡率增加,细胞周期停滞在 G2/M 期。与细胞凋亡相关的 BCL-2/BAX 蛋白表达减少,而 p53 蛋白表达上调。因此,我们已经证明,抑制 SKP2 的表达可以显著阻碍癌细胞的增殖和迁移,使细胞周期停止,并诱导细胞凋亡。
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Silencing of the S-Phase Kinase-Associated Protein 2 Gene (SKP2) Inhibits Proliferation and Migration of Hepatocellular Carcinoma Cells

Abstract

SKP2 gene is an independent prognostic factor in some diseases and a potential oncogene. The molecular mechanism underlying the occurrence and development of hepatoma, and the involvement of the SKP2 in this process remain unclear. Here, in order to study the effect of SKP2 on proliferation, apoptosis and migration of hepatoma cells, we utilized lentivirus-mediated RNA interference technology using short hairpin RNAs (shRNAs) specific for SKP2. It was demonstrated that SKP2 expression was significantly upregulated in 809 hepatocarcinoma tissues compared to 379 normal liver tissues. The survival time of patients with high levels of SKP2 mRNA was shorter than those with low levels, and SKP2 expression was maximal in stage III hepatocellular carcinoma tissues. The effects of SKP2 silencing on proliferation, apoptosis, cell cycle, migration, and the expression of apoptosis proteins in Huh7 and HepG2 cells were evaluated by MTT assay, flow cytometry, colony formation assay, Transwell, and Western blot analysis. SKP2 expression was significantly reduced in stably transfected Huh7 and HepG2 cells, with knockout efficiencies of 95.7 and 85.8%, respectively. The viability, proliferation, and migration of transfected cancer cells were reduced. In these cells, the apoptosis rate was increased, and the cell cycle was arrested in the G2/M phase. The expression of the apoptosis-associated BCL-2/BAX proteins was decreased, while p53 was upregulated. Thus, we have shown that inhibiting the expression of SKP2 can significantly impede cancer cell proliferation and migration, halt the cell cycle, and induce apoptosis.

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来源期刊
Molecular Biology
Molecular Biology 生物-生化与分子生物学
CiteScore
1.30
自引率
8.30%
发文量
78
审稿时长
3 months
期刊介绍: Molecular Biology is an international peer reviewed journal that covers a wide scope of problems in molecular, cell and computational biology including genomics, proteomics, bioinformatics, molecular virology and immunology, molecular development biology, molecular evolution and related areals. Molecular Biology publishes reviews, experimental and theoretical works. Every year, the journal publishes special issues devoted to most rapidly developing branches of physical-chemical biology and to the most outstanding scientists.
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