精准医学杂志 (Jun 2024)
Effect of STX5 on the metastasis of hepatocellular carcinoma and its mechanism
Abstract
Objective To investigate the effect of STX5 on the metastasis of hepatocellular carcinoma (HCC) and its mechanism. Methods Clinical data were collected from 36 patients who were diagnosed with HCC in our hospital from January to December 2015, and the correlation between the expression level of STX5 in tumor tissue and the clinicopathological features of HCC patients was analyzed. Human hepatoma cells MHCC97H were divided into groups A and B and were transfected with negative control plasmid and STX5 overexpression plasmid, respectively. Human hepatoma cells Huh7 were divided into groups C and D and were transfected with negative control lentivirus and STX5 knockdown virus, respectively. Western blot was used to measure the protein expression level of STX5 in groups A, B, C, and D, wound healing was used to measure the migration ability of cells in groups A, B, C, and D, and Transwell assay was used to measure the migration ability of cells in groups A, B, C, and D. Gene ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis were performed for the differentially expressed genes in groups A and B obtained by transcriptomic sequencing, and RT-qPCR was used to measure the levels of the most significantly differentially expressed genes in group A and B. MHCC97H cells were divided into groups E, F, G, and H and were transfected with negative control plasmid, STX5 overexpression plasmid, negative control plasmid+Sarilumab, and STX5 overexpression plasmid+Sarilumab, respectively. Scratch assay was used to measure the migration ability of cells in groups E, F, G, and H, and Transwell assay was used to measure the migration ability of cells in groups E, F, G, and H. Results The expression level of STX5 in tumor tissue was associated body mass index, presence or absence of hepatitis B virus infection, and the number of tumors (P<0.05). Western blot showed that group B had a significantly higher expression level of STX5 than group A, and group C had a significantly higher expression level of STX5 than group D (t=48.86,31.09,P<0.05). Transwell test and wound healing showed that compared with group A, group B had significantly higher ability of cell invasion and percentage of scratch healing, and compared with group D, group C had significantly higher ability of cell migration and percentage of scratch healing (t=7.95-31.09,P<0.05). The GO and KEGG enrichment analyses showed that the differentially expressed genes between groups A and B were mainly enriched in the functions and pathways associated with cell migration and inflammation, and RT-qPCR showed that group B had a significantly higher mRNA expression level of IL-6 than group A (t=23.69,P<0.05). Transwell assay and wound healing showed that group G had significantly lower ability of cell migration and percentage of scratch healing than group E, and group H had significantly lower ability of cell invasion and percentage of scratch healing than group F (t=2.94-24.39,P<0.05). Conclusion STX5 can promote the migration and metastasis of HCC cells by upregulating the mRNA expression level of IL-6.
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