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中华普通外科学文献(电子版)

中华普通外科学文献(电子版) ›› 2024, Vol. 18 ›› Issue (03) : 192 -198. doi: 10.3877/cma.j.issn.1674-0793.2024.03.004

论著

补体B因子在结直肠癌中的表达及临床意义
陈金业1, 凌潜龙1, 朱冰1, 骆杰1,†()   
  1. 1. 233000　蚌埠医学院第一附属医院胃肠外科
  • 收稿日期:2023-12-07 出版日期:2024-06-01
  • 通信作者: 骆杰
  • 基金资助:
    安徽省高等学校自然科学研究项目(KJ2021ZD0090); 蚌埠医学院研究生科研创新项目(Byycx22090)

Expression and clinical significance of complement B factor in colorectal cancer

Jinye Chen1, Qianlong Ling1, Bing Zhu1, Jie Luo1,†()   

  1. 1. Department of Gastrointestinal Surgery, the First Affiliated Hospital of Bengbu Medical College, Bengbu 233000, China
  • Received:2023-12-07 Published:2024-06-01
  • Corresponding author: Jie Luo
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陈金业, 凌潜龙, 朱冰, 骆杰. 补体B因子在结直肠癌中的表达及临床意义[J]. 中华普通外科学文献(电子版), 2024, 18(03): 192-198.

Jinye Chen, Qianlong Ling, Bing Zhu, Jie Luo. Expression and clinical significance of complement B factor in colorectal cancer[J]. Chinese Archives of General Surgery(Electronic Edition), 2024, 18(03): 192-198.

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=0.004 1）。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=0.000 1）。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=0.000 2）。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。

关键词: 结直肠肿瘤, 补体B因子, 计算生物, 迁移, 侵袭
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=0.004 1). 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 stageⅠand Ⅱ were 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.

Key words: Colorectal neoplasms, Complement B factor, Computational biology, Migration, Invasion
图1 研究补体B因子（CFB）在结直肠癌中表达及意义的实验设计流程图
表1 细胞siRNA转染序列
siRNA 正义链序列 反义链序列
阴性对照siRNA 5’-UUCUCCGAACGUGUCACGUTT-3’ 5’-ACGUGACACGUUCGGAGAATT-3’
siRNA1 5’-CACGAAGCAGCUCAAUGAA-3’ 5’-UUCAUUGAGCUGCUUCGUG-3’
siRNA2 5’- GGAGAUAGAAGUAGUCCUA-3’ 5’-UAGGACUACUUCUAUCUCC-3’
siRNA3 5’- GUGCUAGAUGGAUCAGACA-3’ 5’-UGUCUGAUCCAUCUAGCAC-3’
图2 CFB在人体正常细胞、组织中的表达　A为CFB在细胞中的分布；B、C、D分别为CFB在结肠细胞、小肠细胞、不同人体组织中的表达水平
图3 CFB在不同肿瘤中的表达水平　A为CFB在泛癌水平上的表达，* P<0.05，** P<0.01，*** P<0.001；B为结直肠癌组织与非癌组织中CFB表达的免疫组织化学结果（×100）
图4 CFB表达水平与结直肠癌患者预后的关系　A为CFB在结直肠癌组织中的表达，* P<0.05；B为CFB与MKI67表达的相关性；C、D分别为结直肠癌患者的总生存率、无进展生存率
图5 结直肠癌中CFB基因的通路富集　A为生物过程的GO分析；B为细胞组成的GO分析；C为分子功能的GO分析；D为KEGG通路富集分析结果
表2 CFB表达与结直肠癌临床病理特点的关系(例)
项目 阳性 阴性 χ2值 P值
例数 35 28    
性别     0.003 0.955
男 19 15    
女 16 13    
年龄(岁)   7.750 0.005
≥60 29 14    
<60 6 14    
病理分期     4.062 0.044
Ⅰ+Ⅱ 10 15    
Ⅲ+Ⅳ 25 13    
T分期     1.260 0.262
T1+T2 8 10    
T3+T4 27 18    
图6 CFB在结直肠癌组织和癌旁正常组织中的表达情况　A、B分别为免疫组织化学、蛋白质水平检测结果；C为免疫组织化学联合染色评分比较；D为CFB蛋白的Western blotting检测量比较，***P<0.001
图7 CFB对结直肠癌细胞迁移、侵袭的影响　A、B分别为RKO细胞、SW620细胞的Western blotting结果，* P<0.05，** P<0.01
图8 CFB对结直肠癌细胞迁移、侵袭的影响　A、D为RKO细胞Transwell小室实验结果（×40）；B、E为SW620细胞Transwell小室实验结果（×40）；C、F为结直肠癌细胞侵袭实验结果（×200）比较；** P<0.01，*** P<0.001
[1]
Sane S, Srinivasan R, Potts RA, et al. UBXN2A suppresses the Rictor-mTORC2 signaling pathway, an established tumorigenic pathway in human colorectal cancer[J]. Oncogene, 2023, 42(21): 1763-1776.
[2]
徐凤敏, 陈昱伶, 任玲, 等. S1P/S1PR1通路调节补体B因子的表达对食管鳞状细胞癌细胞增殖和迁移的影响[J]. 中国免疫学杂志, 2021, 37(19): 2364-2369.
[3]
Lu Q, Hou Q, Cao K, et al. Complement factor B in high glucose-induced podocyte injury and diabetic kidney disease[J]. JCI Insight, 2021, 6(19): e147716.
[4]
Wu P, Shi J, Sun W, et al. The prognostic value of plasma complement factor B (CFB) in thyroid carcinoma[J]. Bioengineered, 2021, 12(2): 12854-12866.
[5]
Shimazaki R, Takano S, Satoh M, et al. Complement factor B regulates cellular senescence and is associated with poor prognosis in pancreatic cancer[J]. Cell Oncol (Dordr), 2021, 44(4): 937-950.
[6]
Lee MJ, Na K, Shin H, et al. Early diagnostic ability of human complement factor B in pancreatic cancer is partly linked to its potential tumor-promoting role[J]. J Proteome Res, 2021, 20(12): 5315-5328.
[7]
Deng YZ, Yao F, Li JJ, et al. RACK1 suppresses gastric tumorigenesis by stabilizing the β-catenin destruction complex[J]. Gastroenterology, 2012, 142(4): 812-823, e15.
[8]
郭楠楠, 王兴智. 补体B因子与特发性膜性肾病的相关性研究[J]. 哈尔滨医科大学学报, 2022, 56(1): 21-25.
[9]
Di Stefano I, Alì G, Poma AM, et al. New immunohistochemical markers for pleural mesothelioma subtyping[J]. Diagnostics (Basel), 2023, 13(18): 2945.
[10]
Petr V, Thurman JM. The role of complement in kidney disease[J]. Nat Rev Nephrol, 2023, 19(12): 771-787.
[11]
Zanchi C, Locatelli M, Corna D, et al. Liver factor B silencing to cure C3 glomerulopathy: evidence from a mouse model of complement dysregulation[J]. Mol Immunol, 2023, 161: 25-32.
[12]
Akhlaghpour M, Haritunians T, More SK, et al. Genetic coding variant in complement factor B (CFB) is associated with increased risk for perianal Crohn’s disease and leads to impaired CFB cleavage and phagocytosis[J]. Gut, 2023, 72(11): 2068-2080.
[13]
Hakroush S,Tampe B. Association between loss of immune checkpoint programmed cell death protein 1 and active ANCA-associated renal vasculitis[J]. Int J Mol Sci, 2023, 24(3): 2975.
[14]
Robson JL, Thorn R, Williams AC, et al. Gut bacteria promote proliferation in benign S/RG/C2 colorectal tumour cells, and promote proliferation, migration and invasion in malignant HCT116 cells[J]. Sci Rep, 2023, 13(1): 17291.
[15]
Rodriguez-Hernandez I, Maiques O, Kohlhammer L, et al. WNT11-FZD7-DAAM1 signalling supports tumour initiating abilities and melanoma amoeboid invasion[J]. Nat Commun, 2020, 11(1): 5315.
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