敲低SKA1通过凋亡抑制肝细胞癌进展:整合单细胞转录组学与体内和体外验证

IF 5 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
BioFactors Pub Date : 2025-08-28 DOI:10.1002/biof.70044
Qiong Luo, Qianyuan Zhang, Ziying Zheng, Xiaoyan Jiang
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引用次数: 0

摘要

肝细胞癌(HCC)是最常见的原发性肝癌类型。纺锤体和着丝酶相关复合体1 (SKA1)参与有丝分裂的调节,在调节癌症进展中发挥重要作用。因此,本研究旨在探讨SKA1基因下调对HCC的影响。采用生物信息学分析方法预测SKA1在HCC中的表达、SKA1对HCC患者生存率和预后的影响以及SKA1表达与基因突变和免疫细胞浸润的关系。单细胞转录组测序分析用于探究细胞间通讯和分子相互作用。采用CCK-8、创面愈合、Transwell、流式细胞术、qRT-PCR等方法检测SMMC7721细胞的细胞活力、侵袭、迁移、细胞周期、凋亡及SKA1 mRNA表达水平。测量肿瘤体积和重量。Western blot检测SMMC7721细胞及肿瘤组织中SKA1、survivin、Bax、Bad、Bcl-2、caspase-3、caspase-9蛋白的表达水平。生物信息学分析结果显示,SKA1高表达与HCC患者生存率低、预后差有关,并参与了TP53突变和多发免疫细胞浸润。单细胞转录组测序分析证实,恶性细胞与肝细胞、ILC和粒细胞相关。同时,SKA1高表达细胞亚群中多种通路和配体受体对富集,尤其是与凋亡信号相关的蛋白酶激活受体(PARs)通路和MDK-SDC1对。敲低SKA1可降低细胞活力、侵袭和迁移,阻滞细胞S期周期,促进体外细胞凋亡,降低体内肿瘤体积和重量,下调survivin和Bcl-2蛋白表达水平,上调体内和体外caspase 3、caspase 9、Bax和Bad蛋白表达水平。综上所述,敲低SKA1通过促进细胞凋亡信号通路抑制HCC进展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Knocking Down SKA1 Inhibits Hepatocellular Carcinoma Progression via Apoptosis: Integrating Single-Cell Transcriptomics With In Vivo and In Vitro Validation

Knocking Down SKA1 Inhibits Hepatocellular Carcinoma Progression via Apoptosis: Integrating Single-Cell Transcriptomics With In Vivo and In Vitro Validation

Knocking Down SKA1 Inhibits Hepatocellular Carcinoma Progression via Apoptosis: Integrating Single-Cell Transcriptomics With In Vivo and In Vitro Validation

Knocking Down SKA1 Inhibits Hepatocellular Carcinoma Progression via Apoptosis: Integrating Single-Cell Transcriptomics With In Vivo and In Vitro Validation

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer. Spindle- and kinetochore-associated complex 1 (SKA1) participates in the regulation of mitosis, playing an essential role in regulating cancer progression. Therefore, this study aims to explore the effects of knocking down SKA1 on HCC. The bioinformatics analysis approaches were adopted to predict SKA1 expression in HCC, the role of SKA1 on the survival rate and prognosis of HCC patients, and the associations between SKA1 expression and gene mutation and immune cell infiltration. The single-cell transcriptome sequencing analysis was employed to explore the cell–cell communications and molecular interactions. The CCK-8, wound healing, Transwell, flow cytometry, and qRT-PCR approaches were used to determine the cell viability, invasion, migration, cell cycle, apoptosis, and SKA1 mRNA expression level of SMMC7721 cells. The tumor volume and weight were measured. The Western blot was applied to determine the protein expression levels of SKA1, survivin, Bax, Bad, Bcl-2, caspase-3, and caspase-9 in SMMC7721 cells and tumor tissue. The bioinformatics analysis results indicated that highly expressed SKA1 was related to a low survival rate and poor prognosis of HCC patients and was involved in the TP53 mutation and multiple immune cell infiltrations. The single-cell transcriptome sequencing analysis affirmed that malignant cells were associated with hepatocytes, ILC, and granulocytes. Meanwhile, various pathways and ligand-receptor pairs were enriched in the cell subpopulation with high SKA1 expression, especially for the Protease-Activated Receptors (PARs) pathway and MDK-SDC1 pair associated with the apoptosis signaling. Knocking down SKA1 reduced the cell viability, invasion, and migration, arrested the cell cycle in the S period, promoted the apoptosis in vitro, decreased the tumor volume and weight in vivo, and down-regulated the survivin and Bcl-2 protein expression levels and up-regulated the caspase 3, caspase 9, Bax, and Bad in vivo and in vitro. Taken together, knocking down SKA1 inhibited HCC progression by promoting the apoptosis signaling pathway.

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来源期刊
BioFactors
BioFactors 生物-内分泌学与代谢
CiteScore
11.50
自引率
3.30%
发文量
96
审稿时长
6-12 weeks
期刊介绍: BioFactors, a journal of the International Union of Biochemistry and Molecular Biology, is devoted to the rapid publication of highly significant original research articles and reviews in experimental biology in health and disease. The word “biofactors” refers to the many compounds that regulate biological functions. Biological factors comprise many molecules produced or modified by living organisms, and present in many essential systems like the blood, the nervous or immunological systems. A non-exhaustive list of biological factors includes neurotransmitters, cytokines, chemokines, hormones, coagulation factors, transcription factors, signaling molecules, receptor ligands and many more. In the group of biofactors we can accommodate several classical molecules not synthetized in the body such as vitamins, micronutrients or essential trace elements. In keeping with this unified view of biochemistry, BioFactors publishes research dealing with the identification of new substances and the elucidation of their functions at the biophysical, biochemical, cellular and human level as well as studies revealing novel functions of already known biofactors. The journal encourages the submission of studies that use biochemistry, biophysics, cell and molecular biology and/or cell signaling approaches.
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