A Analysis on Expression Significance of BUB1 in Breast Cancer and Intervention of Japonicone A Based on Data-mining from Bioinformatics / 中国实验方剂学杂志

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⁺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.

BUB1 gene is highly expressed in gastric cancer:analysis based on Oncomine database and bioinformatics / 南方医科大学学报

OBJECTIVE@#To investigate the expression of BUB1 gene in gastric cancer.@*METHODS@#Oncomine, GEPIA, BioGPS and Kaplan-Meier Plotter databases were used to analyze the difference of BUB1 gene expression between gastric cancer tissue and normal gastric tissue. The association of BUB1 expression level with the prognosis of gastric cancer patients was also analyzed. The Cancer Cell Line Encyclopedia (CCLE) was explored to analyze the expression of BUB1 in T cells and B cells in gastric cancer patients, and the String database was used to generate the network map of BUB1-related proteins and functional annotation of gene ontology (GO). The related pathways of KEGG were analyzed. Tumor immune assessment resource (TIMER) database was used to analyze the expression of BUB1 in immune infiltration and its effect on prognosis of gastric cancer patients. To further verify the results of gene chip analysis in Oncomine database, we collected 30 pairs of surgical specimens of gastric adenocarcinoma and adjacent tissues from patients admitted to the First Affiliated Hospital of Chengdu Medical College from March, 2018 to July, 2019. The results of BUB1 gene expression in Oncomine database were verified by PCR and immunohistochemistry.@*RESULTS@#Oncomine, GEPIA and BioGPS analyses showed that BUB1 was highly expressed in gastric cancer compared with normal gastric tissue. Kaplan-Meier survival analysis showed that the progression-free survival time (HR=0.52, 95% :0.41-0.67, < 0.05) and the overall survival time (HR=0.67, 95% :0.55-0.82, < 0.05) were prolonged in gastric cancer patients with a high expression of BUB1. Through String data collection, BUB1-related proteins were mainly enriched in 13 cellular components, 4 molecular functions and 12 biological processes, involving 4 signal pathways. TIMER database analysis showed that CD4 T cells and macrophages with high expressions of BUB1 mRNA in the immune microenvironment were associated with a favorable 5-year survival outcome of patients with gastric cancer. In the surgical specimens, real-time quantitative PCR showed that the expression level of BUB1 mRNA was significantly higher in gastric cancer tissues than in the adjacent gastric mucosa tissues, and immunohistochemical results demonstrated positive BUB1 staining in the gastric cancer tissues.@*CONCLUSIONS@#BUB1 gene is highly expressed in gastric cancer. BUB1 may reduce tumor immunosuppression and helps to evaluate the prognosis of patients with gastric cancer.

BUB1 expression is enriched in glioblastoma and promotes tumor proliferationin glioblastoma / 西安交通大学学报(医学版)

Objective: To investigate the functions of mitosiskinase budding uninhibited by benzimidazoles 1 (BUB1) for cell proliferation in glioblastoma (GBM). Methods: We investigated the expression level of BUB1 in GBM cells by analyzing TCGA database. Then Real-time PCR and Western blot were used to explore the role of BUB1 in proliferation of GBM cells via BUB1 silencing using siRNA transfection. Additionally, we investigated the correlation between BUB1 expression and prognosis of GBM patients. Results: BUB1 expression was significantly elevated in GBM patients, while artificial silencing of BUB1 attenuated the proliferation (P=0.007) and cloning formation of U87 cells (P=0.033). At the same time, improved prognosis could be observed in GBM patients with highly expressed BUB1 compared with those with lower BUB1 expression (P=0.040). Conclusion: Increased BUB1 expression promotes the growth and proliferation of GBM cells and is correlated with the poor prognosis of GBM patients.

The role of micro RNAs let7c, 100 and 218 expression and their target RAS, C-MYC, BUB1, RB, SMARCA5, LAMB3 and Ki-67 in prostate cancer

OBJECTIVE: The aim of this study is to verify the expression of proteins that are controlled by miR-let7c, 100 and 218 using immunohistochemistry in tissue microarray representative of localized and metastasized the lymph nodes and bone prostate cancer. METHODS: To verify the expression of proteins that are controlled by miR-let7c (C-MYC, BUB1, RAS) 100 (SMARCA5, RB) and 218 (LAMB3) and cell proliferation (Ki-67) we used immunohistochemistry and computerized image system ImageJ MacBiophotonics in three tissue microarrays representative of localized prostate cancer and lymph node and bone metastases. miRNA expression was evaluated by qRT-PCR using 60 paraffin blocks to construct the tissue microarray representative of localized disease. RESULTS: RAS expression was increased in localized prostate cancer and bone metastases compared to the lymph nodes (p=0.017). RB showed an increase in expression from localized prostate cancer to lymph node and bone metastasis (p=0.036). LAMB3 was highly expressed in localized and lymph node metastases (p<0.001). Cell proliferation evaluated by Ki-67 showed an increase from localized prostate cancer to metastases (p<0.001). We did not found any relationship between C-MYC (p=0.253), BUB1 (p=0.649) and SMARCA5 (p=0.315) protein expression with prognosis or tumor behavior. CONCLUSION: We found that the expression of RAS, RB, LAMB3 and Ki-67 changed in the different stages of prostate cancer. Furthermore, we confirmed the overexpression of the miRNAs let7c, 100 and 218 in localized prostate cancer but failed to show the control of protein expression by the putative controller miRNAs using immunohistochemistry. .

Differential promoter methylation may be a key molecular mechanism in regulating BubR1 expression in cancer cells

The BubR1 mitotic-checkpoint protein monitors proper attachment of microtubules to kinetochores, and links regulation of chromosome-spindle attachment to mitotic-checkpoint signaling. Thus, disruption of BubR1 activity results in a loss of checkpoint control, chromosomal instability caused by a premature anaphase, and/or the early onset of tumorigenesis. The mechanisms by which deregulation and/or abnormalities of BubR1 expression operate, however, remain to be elucidated. In this study, we demonstrate that levels of BubR1 expression are significantly increased by demethylation. Bisulfite sequencing analysis revealed that the methylation status of two CpG sites in the essential BubR1 promoter appear to be associated with BubR1 expression levels. Associations of MBD2 and HDAC1 with the BubR1 promoter were significantly relieved by addition of 5-aza-2'-deoxycytidine, an irreversible DNA methyltransferase inhibitor. However, genomic DNA isolated from 31 patients with colorectal carcinomas exhibited a +84A/G polymorphic change in approximately 60% of patients, but this polymorphism had no effect on promoter activity. Our findings indicate that differential regulation of BubR1 expression is associated with changes in BubR1 promoter hypermethylation patterns, but not with promoter polymorphisms, thus providing a novel insight into the molecular regulation of BubR1 expression in human cancer cells.

Expression of BUB1 gene in trophoblastic cells was depressed by microRNA / 重庆医科大学学报

Objective:To discuss the depressed expression of BUB1 gene in trophoblastic cells by microRNA.Methods:Endogenous BUB1 mRNA level was compared by using quantitative real-time PCR before and after transfected with microRNA plasmid; and the Bub1 protein level was compared by western blot.Results:Ratio of BUB1 mRNA/GAPDH mRNA of control group was 0.1960?0.0688,and after transfected with microRNA recombinant plasmid was 0.1968?0.0689 and 0.1962?0.0670 respectively.Statistical study of samples showed they have no instinct different;otherwise the protein level was significantly decreased after transfected with microRNA plamid.Conclusion:This study established microRNA can control gene expression in mammalian cells at translation level,it is a good foundation to study on the control of gene expression.