补体B因子在结直肠癌中的表达及临床意义

Expression and clinical significance of complement B factor in colorectal cancer

目的基于生物信息学与细胞实验探究补体B因子(CFB)在结直肠癌中的表达及临床意义。方法利用HPA和TIMER 2.0数据库分析CFB在结直肠癌组织中的表达;GEPIA数据库分析CFB与肿瘤生长因子MKI67在结直肠癌中表达的关联;Kaplan-Meier Plotter数据库分析CFB mRNA的表达水平与结直肠癌患者生存期的关系;基因富集通路预测CFB对结直肠癌潜在的生物学功能;免疫组织化学(IHC)和Western blotting验证CFB在结直肠癌组织中的表达;采用siRNA转染,利用细胞划痕、Transwell小室实验探究CFB对结直肠癌细胞迁移、侵袭的影响。结果HPA和TIMER2.0数据库显示CFB在结直肠癌组织中高表达(P<0.001)。GEPIA数据库显示结直肠癌组织中CFB表达较高(P<0.05),且与MKI67表达呈正相关(P=0.0041)。Kaplan-Meier Plotter生存分析表明,低表达的CFB在结直肠癌中预示更差的总生存期(P=0.015)和进展后生存期(P=0.048)。基因富集结果显示,CFB可能参与多个结直肠癌进展通路。IHC显示,CFB在结直肠癌组织中阳性染色强度高于正常组织,且在TNMⅠ、Ⅱ期中的表达弱于Ⅲ、Ⅳ期(P=0.0001)。Western blotting显示在10组结直肠癌及配对癌旁组织中,CFB在结直肠癌组织中表达更高(P=0.0002)。细胞划痕、Transwell小室结果显示,CFB促进RKO、SW620细胞的迁移、侵袭能力(P<0.01)。结论CFB在结直肠癌组织中高表达,并与不良预后相关,而RKO、SW620细胞的迁移、侵袭能力可被低表达的CFB抑制,提示其可作为结直肠癌患者预后的潜在标志物。

Objective To explore the expression and clinical significance of complement B factor(CFB)in colorectal cancer(CRC)based on bioinformatics and cell experiments.Methods HPA and TIMER 2.0 databases were used to analyze the expression of CFB in CRC tissue;GEPIA database analysis of the association between CFB and MKI67 expression in CRC.Kaplan-Meier Plotter database was used to analyze the relationship between the expression level of CFB mRNA and the survival of CRC patients;Gene enrichment pathway to predict the potential biological function of CFB in CRC;IHC and Western blotting validation of CFB expression in CRC tissue.The effects of CFB on migration and invasion of CRC cells were investigated by siRNA transfection,cell scratch and Transwell assay.Results The bioinformatics database showed that CFB was highly expressed in CRC tissues(P<0.001).The GEPIA database showed that CFB expression was higher in CRC tissues(P<0.05)and positively correlated with MKI67 expression(P=0.0041).Kaplan-Meier Plotter survival analysis showed that low expression of CFB predicted poorer overall survival(P=0.015)and progression survival(P=0.048)in CRC.The enrichment results of CFB gene indicated that it might be involved in multiple progression pathways of CRC.IHC showed that the positive staining intensity of CFB in CRC tissue was higher than that in normal tissue,and TNM stageⅠandⅡwere weaker than Ⅲ and Ⅳ (P=0.000 1). Western blotting showed that in 10 cases of CRC and paired adjacent tissues, CFB was more expressed in cancer tissues (P=0.000 2). Cell scratch and Transwell cell results showed that CFB promoted the migration and invasion ability of RKO and SW620 cells (P<0.01). Conclusion CFB is highly expressed in CRC tissue and is associated with prognosis, while the migration and invasion ability of RKO and SW620 cells can be inhibited by low expression of CFB, indicating that it can serve as a potential prognostic marker for CRC.