BACKGROUND Esophageal carcinoma(EC)is one of the most prevalent cancers in human populations worldwide.Baitouweng decoction is one of the most important Chinese medicine formulas,with the potential to treat cancer.AIM...BACKGROUND Esophageal carcinoma(EC)is one of the most prevalent cancers in human populations worldwide.Baitouweng decoction is one of the most important Chinese medicine formulas,with the potential to treat cancer.AIM To investigate the role and mechanism of Baitouweng decoction on EC cells.METHODS Differentially expressed genes(DEGs)in EC tissues and normal tissues were screened by the cDNA microarray technique and by bioinformatics methods.The target genes of microRNAs were predicted based on the TargetScan database and verified by dual luciferase gene reporter assay.We used Baitouweng decoction to intervene EC cells,and detected the activity of EC9706 and KYSE150 cells by the MTT method.Cell cycle and apoptosis were measured by flow cytometry.The expression of BUB1 mRNA and miR-495-3p was measured by qRT-PCR.The protein levels of BUB1,STAT3,p-STAT3,CCNB1,CDK1,Bax,Caspase3,and Caspase9 were measured by Western blot analysis.The migration and invasion abilities of the cells were measured by wound-healing assay and Transwell invasion assay,respectively.RESULTS DEGs identified are involved in biological processes,signaling pathways,and network construction,which are mainly related to mitosis.BUB1 was the key hub gene,and it is also a target gene of miR-495-3p.Baitouweng decoction could upregulate miR-495-3p and inhibit BUB1 expression.In vitro experiments showed that Baitouweng decoction significantly inhibited the migration and invasion of EC cells and induced apoptosis and G2/M phase arrest.After treatment with Baitouweng decoction,the expression of Bax,Caspase 3,and Caspase 9 in EC cells increased significantly,while the expression of BUB1,CCNB1,and CDK1 decreased significantly.Moreover,the STAT3 signaling pathway may play an important role in this process.CONCLUSION Baitouweng decoction has a significant inhibitory effect on EC cell growth.BUB1 is a potential therapeutic target for EC.Further analysis showed that Baitouweng decoction may inhibit the growth of EC cells by upregulating miR-495-3p targeting the BUB1-mediated STAT3 signal pathway.展开更多
目的研究胃癌组织中有丝分裂关卡基因BUB1和BUBR1的表达。方法采用半定量RT-PCR技术检测39例胃癌组织及正常胃组织中BUB1及BUBR1基因的表达。结果胃癌组织中BUB1mRNA/β-actin mRNA和BUBR1mRNA/β-actin mRNA拷贝数比值均明显高于正常...目的研究胃癌组织中有丝分裂关卡基因BUB1和BUBR1的表达。方法采用半定量RT-PCR技术检测39例胃癌组织及正常胃组织中BUB1及BUBR1基因的表达。结果胃癌组织中BUB1mRNA/β-actin mRNA和BUBR1mRNA/β-actin mRNA拷贝数比值均明显高于正常胃组织(1.84±0.43 vs. 0.86±0.44和2.00±1.36 vs. 0.96±0.60)(P<0.05)。结论 BUB1基因和BUBR1基因在胃癌中高表达可能与癌细胞的异常增殖有关。展开更多
基金Supported by National Natural Science Foundation of China,No.81550014Henan Provincial Science and Technology Research Project,No.222102310187 and No.242102310582Henan Province Postdoctoral Research Project,No.202103092.
文摘BACKGROUND Esophageal carcinoma(EC)is one of the most prevalent cancers in human populations worldwide.Baitouweng decoction is one of the most important Chinese medicine formulas,with the potential to treat cancer.AIM To investigate the role and mechanism of Baitouweng decoction on EC cells.METHODS Differentially expressed genes(DEGs)in EC tissues and normal tissues were screened by the cDNA microarray technique and by bioinformatics methods.The target genes of microRNAs were predicted based on the TargetScan database and verified by dual luciferase gene reporter assay.We used Baitouweng decoction to intervene EC cells,and detected the activity of EC9706 and KYSE150 cells by the MTT method.Cell cycle and apoptosis were measured by flow cytometry.The expression of BUB1 mRNA and miR-495-3p was measured by qRT-PCR.The protein levels of BUB1,STAT3,p-STAT3,CCNB1,CDK1,Bax,Caspase3,and Caspase9 were measured by Western blot analysis.The migration and invasion abilities of the cells were measured by wound-healing assay and Transwell invasion assay,respectively.RESULTS DEGs identified are involved in biological processes,signaling pathways,and network construction,which are mainly related to mitosis.BUB1 was the key hub gene,and it is also a target gene of miR-495-3p.Baitouweng decoction could upregulate miR-495-3p and inhibit BUB1 expression.In vitro experiments showed that Baitouweng decoction significantly inhibited the migration and invasion of EC cells and induced apoptosis and G2/M phase arrest.After treatment with Baitouweng decoction,the expression of Bax,Caspase 3,and Caspase 9 in EC cells increased significantly,while the expression of BUB1,CCNB1,and CDK1 decreased significantly.Moreover,the STAT3 signaling pathway may play an important role in this process.CONCLUSION Baitouweng decoction has a significant inhibitory effect on EC cell growth.BUB1 is a potential therapeutic target for EC.Further analysis showed that Baitouweng decoction may inhibit the growth of EC cells by upregulating miR-495-3p targeting the BUB1-mediated STAT3 signal pathway.
文摘目的研究胃癌组织中有丝分裂关卡基因BUB1和BUBR1的表达。方法采用半定量RT-PCR技术检测39例胃癌组织及正常胃组织中BUB1及BUBR1基因的表达。结果胃癌组织中BUB1mRNA/β-actin mRNA和BUBR1mRNA/β-actin mRNA拷贝数比值均明显高于正常胃组织(1.84±0.43 vs. 0.86±0.44和2.00±1.36 vs. 0.96±0.60)(P<0.05)。结论 BUB1基因和BUBR1基因在胃癌中高表达可能与癌细胞的异常增殖有关。
