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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年01期

页码:

56-64

栏目:

临床医学

出版日期:

2024-01-01

文章信息/Info

Title:

Prognosis of patients with bladder cancer predicted by biomarker associated with anoikis

文章编号:

1006-8147(2024)01-0056-09

作者:

朱 亮; 陈业刚

（天津医科大学第二医院泌尿外科，天津300211）

Author(s):

ZHU Liang; CHEN Yegang

（Department of Urology，Tianjin Institute of Urology，The Second Hospital，Tianjin Medical University，Tianjin 300211，China）

关键词:

失巢凋亡; 膀胱癌; 肿瘤微环境; 预后模型; 免疫疗法

Keywords:

anoikis; bladder cancer; tumor microenvironment; prognostic model; immunotherapy

分类号:

R694+.7

DOI:

10.20135/j.issn.1006-8147.2024.01.0056

文献标志码:

A

摘要:

目的：探讨失巢凋亡相关基因在膀胱癌中的作用和预后价值。方法：从Genecards 数据库和Harmonizome数据库获取失巢凋亡相关基因。Cox和LASSO回归用来得到具有预后价值的基因并构建风险预后模型。列线图、校准图、累积危险曲线和决策曲线用来验证模型准确性。研究不同风险组间在生存预后及免疫景观上的差异。采用实时定量PCR测定人正常膀胱细胞SV-HUV-1、T24、J82、5637膀胱癌细胞系中F10表达水平，将构建的含有全长的pcDNA3.1 质粒及阴性对照质粒瞬时转染T24细胞； 采用CCK-8、集落刺激实验检测F10高表达对细胞增殖能力的影响； 利用Transwell实验检测 PCA3 高表达对细胞侵袭能力的影响。结果：获得9个失巢凋亡相关基因（DNMT1、F10、FASN、PDGFRA、SATB1、MSLN、MYC、CALR、CSPG4）用来构建预后模型。列线图、校准图、累积危险曲线和决策曲线显示，模型能较好的评估膀胱癌患者的预后。根据风险评分分组，低危组患者的总生存期（OS）明显长于高危组。免疫微环境中，CD8+ T细胞（P=0.015）、M0巨噬细胞（P=0.005）、M2巨噬细胞（P =0.05）和中性粒细胞（P=0.009）呈现出较大差异。高危组与低危组间免疫评分具有统计学意义（P＜0.001）。经外部验证证实失巢凋亡基因与膀胱癌的预后相关。以F10为例，RT-qPCR结果表明T24细胞系相对表达水平低于正常膀胱癌上皮（t=3.031，P＜0.05）；CCK-8实验中，高表达F10组24、48、72 h增殖活性低于对照组（t=2.578、5.528、19.25，均P＜0.001）；集落刺激形成实验表明高表达F10组细胞克隆形成数低于对照组（t=10.570，P＜0.001）；Transwell迁移侵袭实验中，高表达F10组细胞迁移及侵袭数目均低于对照组（t=8.115，P＜0.005）。结论：共获得9个失巢凋亡相关基因，其可以作为潜在的治疗靶点应用于临床。

Abstract:

Objective： To explore the role and prognostic value of anoikis-related genes（ARGs） in bladder cancer. Methods： ARGs were obtained from the Genecards and Harmonizome databases. Cox and LASSO regressions were used to obtain genes with prognostic value and to construct risk prognostic models. The accuracy of the model was validated using nomogram， calibration plots， cumulative hazard curves and decision curve analysis. Further， we explored differences in survival prognosis and immune landscapes among different risk groups. Finally，real-time quantitative PCR was used to determine the expression level of F10 in normal human bladder cells SV-HUV-1 and T24， J82， 5637 bladder cancer cell lines，and the constructed plasmid containing full length pcDNA3.1 and the negative control plasmids were transiently transfected into T24 cells；CCK-8 and colony formation assay were applied to evaluate the influence of high F10 expression on cell proliferation． Transwell assay was used to test the effect of PCA3 on cell migration. Results： Nine ARGs（DNMT1， F10， FASN， PDGFRA， SATB1， MSLN， MYC， CALR， CSPG4） were obtained for use in constructing prognostic models. The nomogram， calibration plots， cumulative hazard curves， and decision curves showed that the model was able to evaluate the prognosis of patients with bladder cancer reasonably well. By grouping patients based on risk scores， patients in the low-risk group had significantly longer overall survival（OS） than those in the high-risk group.In addition， CD8+ T cells（P=0.015）， M0 macrophages（P=0.005）， M2 macrophages （P=0.05）， and neutrophils（P=0.009） showed large differences in the immune microenvironment. The immune scores were statistically significant between the high-risk and low-risk groups（P<0.001）. At last， the association of ARGs with the prognosis of bladder cancer was further confirmed by external validation.Taking F10 as an example， RT-qPCR results showed that the relative expression level of T24 cell line was lower than that of normal bladder cancer epithelium cells（t=3.031， P<0.05）. In the CCK-8 assay， the proliferative activities at 24， 48， and 72 h in the high F10-expressing group were lower than those in the control group（t=2.578， 5.528， 19.25， all P<0.001）； the colony-forming assays showed that the number of cell clone formations in the high F10-expressing group was lower than that in the control group（t=10.570， P<0.001）； and in the transwell migration and invasion assay， the number of cell migration and invasion in the high F10-expressing group was also lower than that in the control group（t=8.115， P<0.005）. Conclusion： A total of 9 ARGs are obtained， which are available for clinical application as potential therapeutic targets.

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备注/Memo

备注/Memo:

基金项目 天津市卫健委一般项目（2021171）
作者简介 朱亮（1998-），男，硕士在读，研究方向：泌尿外科；通信作者：陈业刚，E-mail：yegangchen@tmu.edu.cn。

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更新日期/Last Update: 2024-01-01