GJB4 Gene Expression in Relation to Clinical and Pathological Features of Pancreatic Cancer Patients
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摘要:
目的 探究GJB4基因在胰腺癌组织中的表达情况，并研究其与临床病理之间的相关性。 方法 运用生物信息学分析癌症基因组图谱（TCGA）和基因型组织表达（GTEx）的RNA测序数据集，对胰腺癌和癌旁中GJB4 mRNA的表达水平进行分析。收集整理2019年1月至2023年12月昆明医科大学第二附属医院病理科120例胰腺癌和40例癌旁样本，采用免疫组化染色方法检测GJB4蛋白在这两者之间的表达差异。使用RT-qPCR方法检测在4个胰腺癌细胞系中GJB4的表达水平。使用单因素和多因素Cox回归和Kaplan-Meier曲线对GJB4和胰腺癌患者的临床病理数据进行分析。此外，运用DAVID功能注释生物信息学和GSEA富集分析GJB4在胰腺癌中的相关通路。 结果 GJB4 mRNA的表达水平在胰腺癌中表达高于癌旁组织，且GJB4的高表达与患者的预后不良差异有统计学意义（P < 0.05）。免疫组化检测显示，GJB4蛋白在胰腺癌组织中呈棕黄色颗粒状，主要在细胞质和细胞膜中表达，GJB4蛋白在胰腺癌中的表达上调（P < 0.05）。RT-qPCR检测结果显示，在4个胰腺癌细胞系中有3个胰腺癌细胞系表达上调（P < 0.05）。COX回归多因素分析发现，GJB4基因在胰腺癌患者预后生存中扮演了独立的危险因素角色。GO富集分析结果发现GJB4与胰腺癌跨膜转运、离子通道、信号释放、膜电位调节等过程相关。GSEA分析发现GJB4在Wnt/β-catenin 信号通路中富集。 结论 在胰腺癌组织中，GJB4的高表达水平与患者的临床病理特征密切相关，这可能预示着患者不良预后。GJB4可能参与调控胰腺癌的Wnt/β-catenin信号通路，并有望成为胰腺癌预后的潜在生物标记物之一。 Abstract:Objective To investigate the expression of GJB4 gene in pancreatic cancer tissue and its correlation with clinicopathology. Methods The expression levels of GJB4 mRNA in pancreatic cancer and adjacent cancer tissues were analyzed using bioinformatics to analyze the Cancer Genome Atlas （TCGA） and Genotype-Tissue Expression （GTEx） RNA sequencing datasets. A total of 120 pancreatic cancer samples and 40 adjacent cancer samples from the Pathology Department of The Second Affiliated Hospital of Kunming Medical University from January 2019 to December 2023 were collected and sorted. Immunohistochemistry staining method was used to detect the expression difference of GJB4 protein between the two groups. RT-qPCR method was used to detect the expression levels of GJB4 in four pancreatic cancer cell lines. Univariate and multivariate Cox regression and Kaplan-Meier curves were used to analyze the clinical pathological data of GJB4 and pancreatic cancer patients. DAVID functional annotation bioinformatics and GSEA enrichment analysis were used to explore the relevant pathways of GJB4 in pancreatic cancer. Results The expression level of GJB4 mRNA in pancreatic cancer was higher than that in adjacent tissues, and the high expression of GJB4 was significantly associated with poor prognosis of patients （P < 0.05）. Immunohistochemical analysis showed that GJB4 protein was brown-yellow granular in pancreatic cancer tissues, mainly expressed in cytoplasm and cell membrane, and GJB4 protein expression was up-regulated in pancreatic cancer （P < 0.05）. The RT-qPCR test results showed that out of 4 pancreatic cancer cell lines, 3 of them had upregulated expression （P < 0.05）. COX regression analysis showed that GJB4 gene was an independent risk factor in the prognosis of pancreatic cancer patients. The results of GO enrichment analysis showed that GJB4 was related to the transmembrane transport, ion channel, signal release and membrane potential regulation of pancreatic cancer. GSEA analysis showed that GJB4 was enriched in the Wnt/β-catenin signaling pathway. Conclusion In pancreatic cancer, the high expression level of GJB4 is closely related to the clinicopathological features of the patients, which may predict the poor prognosis of the patients. GJB4 may be involved in regulating the Wnt/β-catenin signaling pathway of pancreatic cancer, and is expected to be one of the potential biomarkers of pancreatic cancer prognosis. -
Key words:
- Pancreatic cancer /
- GJB4 /
- Clinicopathological features /
- Prognosis /
- Biomarkers
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图 1 人类不同癌症中GJB4的表达水平
A：UCSC XENA用于检测对TCGA和GTEx数据中不同肿瘤中GJB4的表达水平；B：GJB4表达在胰腺癌和正常组织中的表达差异；C: GJB4在胰腺癌中的ROC曲线。
Figure 1. Expression levels of GJB4 in different human cancers
2 胰腺癌患者GJB4表达与临床特征之间的相关性
A～H：GJB4的高表达与不同临床患者的T分期、N分期、病理分期、肿瘤直径、组织分级、OS事件、DSS事件、PFI事件的表达水平；I～L：GJB4的高表达与患者患有糖尿病、胰腺炎、病理分级、组织分期的相关性。
2. Correlation between GJB4 expression and clinical features in patients with pancreatic cancer
图 3 GJB4在胰腺癌中的诊断价值
A：预测胰腺癌患者总生存率的列线图；B：胰腺癌患者相关的森林图。
Figure 3. Diagnostic value of GJB4 in pancreatic cancer
图 4 胰腺癌中GJB4高表达和低表达的预后意义进行Kaplan-Meier生存曲线分析
A：使用UALCAN数据库评估胰腺癌患者的总生存率；B：Kaplan-Meier评估胰腺癌患者中TCGA患者的总生存率。
Figure 4. The prognostic significance of high and low expression of GJB4 in pancreatic cancer was analyzed by Kaplan-Meier survival curve
图 5 GJB4在自己临床样本中免疫组化表达（IHC×100）
A：GJB4在胰腺癌旁组织中表达情况；B：GJB4在胰腺癌组织中呈现低表达水平；C：GJB4在胰腺癌组织中呈现高表达水平。
Figure 5. GJB4 was expressed immunohistochemically in their own clinical samples （IHC×100）
图 6 GJB4免疫组化高低表达构建的K-M曲线
Figure 6. The K-M curve of high and low immunohistochemical expression of GJB4 was constructed
图 7 GJB4在胰腺癌细胞系和正常胰腺细胞系中的表达情况
与HPDE6-C7比较，*P < 0.05，***P < 0.001。
Figure 7. Expression of GJB4 in pancreatic cancer cell lines and normal pancreatic cell lines
图 8 GJB4相关基因的GJB4功能聚类和相互作用网络分析
A：胰腺癌中与GJB4呈正相关和负相关的前50个基因；B：胰腺癌中GJB4相关基因的GO通路分析；C：胰腺癌中GJB4相关基因的KEGG通路分析；D～E：GJB4相关基因的GESA富集通路分析。
Figure 8. GJB4 functional clustering and interaction network analysis of GJB4-related genes
表 1 单变量和多变量COX回归分析确定预后因素
Table 1. Univariate and multivariate COX regression analyses identified prognostic factors
特征 总数（n） 单变量分析 多变量分析 HR （95%CI） P HR（95%CI） P T 分期 176 T1&T2 31 Reference T3&T4 145 2.023 （1.072～3.816） 0.030* 1.423 （0.724～2.800） 0.307 N分期 173 N0 50 Reference N1 123 2.154 （1.282～3.618） 0.004* 1.832 （1.053～3.187） 0.032* 病理分期 175 I期&I期I 167 Reference II期I& IV期 8 0.673 （0.212～2.135） 0.501 性别 178 女 80 Reference 男 98 0.809 （0.537-1.219） 0.311 组织学分级 176 G1&G4 33 Reference G2&G3 143 2.096 （1.131～3.884） 0.019* 1.449 （0.772～2.719） 0.248 年龄（岁） 178 ≤65 93 Reference >65 85 1.290 （0.854～1.948） 0.227 GJB4 178 1.405 （1.204～1.639） <0.001* 1.243 （1.044～1.480） 0.015* *P < 0.05。 
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表 2 GJB4在胰腺癌组织和癌旁组织中的差异表达
Table 2. Differential expression of GJB4 in pancreatic and paracancer tissues
类型 胰腺
癌旁
组织胰腺癌
组织χ2 95%CI 效应值 P 低表达 32 82 8.21 （0.75，2.13） 0.45 0.003* 高表达 8 38 1.80 （0.67，2.28） 0.50 0.023* 汇总 40 120 *P < 0.05。
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[1] Bray F,Ferlay J,Soerjomataram I,et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin,2018,68(6):394-424. doi: 10.3322/caac.21492 [2] Siegel R L,Miller K D,Wagle N S,et al. Cancer statistics,2023[J]. CA-Cancer J Clin,2023,73(1):17-48. doi: 10.3322/caac.21763 [3] Xia C,Dong X,Li H,et al. Cancer statistics in China and United States,2022: Profiles,trends,and determinants[J]. Chinese Med J-Peking,2022,135(5):584-590. doi: 10.1097/CM9.0000000000002108 [4] Ushio J,Kanno A,Ikeda E,et al. Pancreatic ductal adenocarcinoma: Epidemiology and risk factors[J]. Diagnostics (Basel),2021,11(3):562. doi: 10.3390/diagnostics11030562 [5] Hue J J,Sugumar K,Markt S C,et al. Facility volume-survival relationship in patients with early-stage pancreatic adenocarcinoma treated with neoadjuvant chemotherapy followed by pancreatoduodenectomys[J]. Surgery,2021,170(1):207-214. doi: 10.1016/j.surg.2020.12.003 [6] 冯永,贺定华,夏昆,等. 角皮病致病基因GJB4在遗传性耳聋中的研究[J]. 中华耳科学杂志,2005,3(4):260-262. [7] Dai S,Wang H,Lin Z,et al. Novel and recurrent mutations in GJB3 and GJB4 cause erythrokeratodermia variabilis et progressiva[J]. Indian Journal of Dermatology,2020,86(1):87-90. [8] Kokotas H,Papagiannaki K,Grigoriadou M,et al. Erythrokeratodermia variabilis: Report of two cases and a novel missense variant in GJB4 encoding connexin 30.3[J]. Eur J Dermatol,2012,22(2):182-186. doi: 10.1684/ejd.2011.1617 [9] Consortium G. The genotype-tissue expression (GTEx) project[J]. Nature Genetics,2013,45(6):580-585. doi: 10.1038/ng.2653 [10] Weinstein J N,Collisson E A,Mills G B,et al. The cancer genome atlas pan-cancer analysis project[J]. Nature Genetics,2013,45(10):1113-1120. doi: 10.1038/ng.2764 [11] Zhao K,Si Y,Sun L,. et al. Efficacy and toxicity of re-irradiation for esophageal cancer patients with locoregional recurrence: A retrospective analysis[J]. Radiation Oncology,2020,15(1):243. doi: 10.1186/s13014-020-01685-2 [12] Kampf C. A Human protein atlas[J]. Journal of Proteomics & Bioinformatics,2008,S2(1):45-46. [13] El-Sabban M E,Pauli B U. Cytoplasmic dye transfer between metastatic tumor cells and vascular endothelium[J]. J Cell Biol,1991,115(5):1375-1382. doi: 10.1083/jcb.115.5.1375 [14] 张孟岳,赵东强. 间隙连接蛋白家族在癌症中作用及其机制的研究进展[J]. 中国病理生理杂志,2023,39(4):747-754. [15] 张桢,田霄峰,陈杰,等. 表皮生长因子通过间隙连接蛋白调控小鼠卵母细胞成熟的研究[J]. 生殖医学杂志,2024,33(5):618-626. [16] Yan,Q,Feng,Z,Jiang,B,et al. Biological functions of connexins in the development of inflammatory bowel disease[J]. Scand J Gastroentero,2024,59(2):142-149. doi: 10.1080/00365521.2023.2267713 [17] Grek C L,Rhett J M,Bruce J S,et al. Connexin 43,breast cancer tumor suppressor: Missed connections[J]. Cancer Lett,2016,374(1):117-126. doi: 10.1016/j.canlet.2016.02.008 [18] Lin Y P,Wu J I,Tseng C W,et al. Gjb4 serves as a novel biomarker for lung cancer and promotes metastasis and chemoresistance via Src activation[J]. Oncogene,2019,38(6):822-837. doi: 10.1038/s41388-018-0471-1 [19] Liu G Y ,Pang Y ,Zhang Y J ,et al. GJB4 promotes gastric cancer cell proliferation and migration via Wnt/CTNNB1 pathway[J]. Ott Vol,2019,12(2): 6745–6755. [20] 黄孝彬,谢梦忆,刘星宇,等. CTNND1通过Wnt/β-catenin信号通路调控胰腺癌细胞增殖、迁移和侵袭[J]. 现代肿瘤医学,2024,32(5):818-825. [21] Parsons M J,Tammela T,Dow L E. WNT as a driver and dependency in cancer[J]. Cancer Discov,2021,11(10):2413-2429. doi: 10.1158/2159-8290.CD-21-0190 [22] 张鹏,陈昆,宋登辉,等. 麦冬皂苷介导Wnt/β-catenin通路对大鼠肝部分切除后肝再生的影响[J]. 中华实用诊断与治疗杂志,2024,38(6):552-556. [23] Alganatay,C,Balbasi,E,Tuncbag,N,et al. SETD3 regulates endoderm differentiation of mouse embryonic stem cells through canonical Wnt signaling pathway[J]. Faseb J,2024,38(4):e23463. doi: 10.1096/fj.202301883R [24] 陈小丽,田小容,黄晓东,等. 肿瘤抑制基因7L通过Wnt/β-catenin信号通路抑制胰腺癌细胞生长的作用机制[J]. 临床内科杂志,2024,41(2):116-119. [25] 陈影,庄蕾,张丹红,等. 低氧微环境通过TGFBI调控Wnt/β-catenin通路介导胰腺癌化疗耐药及机制研究[J]. 现代肿瘤医学,2024,32(1):42-46. -
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