ABSTRACT
Objective:To investigate the expression of the cell cycle mitotic spindle checkpoint serinel/threonine kinase （BUB1） gene in breast cancer and its relationship with the prognosis，and further explore the intervention effect of japonicone A on BUB1 gene in breast cancer cells. Method:Through Oncomine，GEPIA，GEO database，bc-GenExMiner v4.5 and Kaplan-Meier Plotter database， in-depth mining was conducted for BUB1 gene expression-related data，in order to explore the difference in the expression of BUB1 gene in breast cancer tissues and normal breast tissues and its relationship with patient prognosis. Encyclopedia of Cancer Cell Lines （CCLE） was used to analyze the expression of BUB1 in T cells and B cells of breast cancer tissues，STRING database was used to draw BUB1-related protein network diagram and gene ontology（GO） function annotation and analyze relevant pathways in Kyoto Encyclopedia of Genes and Genomes （KEGG）. The tumor immune assessment resource（TIMER） database was used to analyze the expression of BUB1 in immune infiltrates and its impact on the survival and prognosis of patients with gastric cancer. Finally， the effect of japonicone A on the expression level of BUB1 gene in breast cancer cells was further analyzed. Result:①The mRNA level of BUB1 in breast cancer tissues was significantly higher than that of normal tissue samples， but the expression levels of BUB1 in breast cancer patients with different molecular subtypes varied. ② Increased expression of BUB1 could lead to longer distant metastasis-free survival（DMFS），overall survival（OS），and recurrence free survival（RFS） in luminal A subtypes. ③ BUB1 was positively correlated with structural maintenance of chro-mosome 4（SMC4） mRNA expression level， and might be interacted with 10 proteins， such as NUF2， with the strongest interaction relationship. ④ The high expression of BUB1 mRNA in CD8<sup>+ </sup>T cells and neutrophils in breast invasive carcinoma（BRCA） and BRCA-basal had a better two-year survival prognosis than the low expression group；the low expression of BUB1 mRNA in B cells in BRCA-Her2 had a better two-year survival prognosis than the high expression group. ⑤ GEO database analysis data set GSE85871 found that japonicone A could down-regulate the expression of BUB1 gene in breast cancer MCF7 cells. Conclusion:BUB1 gene is highly expressed in breast cancer tissues and related to the prognosis of breast cancer and immune cell infiltration. This may be a new potential therapeutic target for japonicone A to intervene breast cancer cells.
ABSTRACT
Objective: To observe the effect of the (p)ppGpp synthase gene (relA) on the heteroresistance of colistin against Acinetobacter baumannii. Methods: The relA gene in Acinetobacter baumannii ATCC19606 was knocked out by Red homologous recombination technique. The biofilm formation of Acinetobacter baumannii was observed by crystal violet staining. The change of heterogeneous colonies of Acinetobacter baumannii under the action of colistin was detected by population analysis profiles (PAP) and the heterogeneity was calculated. The killing curve was used to detect the formation of persistent bacteria in Acinetobacter baumannii under the action of colistin. Results: The relA gene in Acinetobacter baumannii was successfully knocked out, and the relA knockout strain ATCC19606-ΔrelA was obtained. After relA gene knockout, the biofilm formation of Acinetobacter baumannii decreased significantly. After relA gene knockout, Acinetobacter baumannii significantly reduced the heterogeneous colonies and persistent bacteria formation under the action of colistin. Conclusion: The bacterial stringent reaction (p) ppGpp synthase relA may be an important factor affecting the heteroresistance of Acinetobacter baumannii to colistin.