Knocking Down SKA1 Inhibits Hepatocellular Carcinoma Progression via Apoptosis: Integrating Single-Cell Transcriptomics With In Vivo and In Vitro Validation
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引用次数: 0
Abstract
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.
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.