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中华临床医师杂志(电子版) ›› 2024, Vol. 18 ›› Issue (11) : 1044 -1053. doi: 10.3877/cma.j.issn.1674-0785.2024.11.011

基础研究

KPNB1 在膀胱癌中的表达及其对膀胱癌细胞增殖和迁移能力的影响
嵇宏声1, 魏万顷1, 邱建国1, 王留成1, 姜福金2, 张先云2, 王苏贵2,()   
  1. 1.223400 江苏淮安,南京医科大学康达学院附属涟水人民医院泌尿外科
    2.223002 江苏淮安,徐州医科大学附属淮安医院泌尿外科
  • 收稿日期:2024-09-07 出版日期:2024-11-15
  • 通信作者: 王苏贵
  • 基金资助:
    江苏省第十六批“六大人才高峰”项目资助(2019-WSW-218)徐州医科大学附属医院发展基金资助项目(XYFY202243,XYFM202245)淮安市自然科学基金资助项目(HAB201730,HAB202117)淮安市基础研究计划(联合专项)卫生健康类科研项目(HABL2023094)

Expression of KPNB1 in bladder cancer and its role in proliferation and migration of bladder cancer cells

Hongsheng Ji1, Wanqing Wei1, Jianguo Qiu1, Liucheng Wang1, Fujin Jiang2, Xianyun Zhang2, Sugui Wang2,()   

  1. 1.Department of Urology, Lianshui People’s Hospital of Kangda College Affiliated to Nanjing Medical University, Huai’an 223400, China
    2.Department of Urology, Huai’an Hospital Affiliated with Xuzhou Medical University, Huai’an 223002, China
  • Received:2024-09-07 Published:2024-11-15
  • Corresponding author: Sugui Wang
引用本文:

嵇宏声, 魏万顷, 邱建国, 王留成, 姜福金, 张先云, 王苏贵. KPNB1 在膀胱癌中的表达及其对膀胱癌细胞增殖和迁移能力的影响[J/OL]. 中华临床医师杂志(电子版), 2024, 18(11): 1044-1053.

Hongsheng Ji, Wanqing Wei, Jianguo Qiu, Liucheng Wang, Fujin Jiang, Xianyun Zhang, Sugui Wang. Expression of KPNB1 in bladder cancer and its role in proliferation and migration of bladder cancer cells[J/OL]. Chinese Journal of Clinicians(Electronic Edition), 2024, 18(11): 1044-1053.

目的

基于生物信息学和细胞实验分析核转运蛋白KPNB1(Karyopherin β1)在膀胱癌中的表达及其作用机制探索。

方法

从癌症基因图谱数据库(TCGA)、基因综合表达(GEO)数据库和人类蛋白组图谱(THPA)获取KPNB1 在膀胱癌组织及正常膀胱组织当中的mRNA 与蛋白表达水平,分析KPNB1 与膀胱癌患者临床病理特征的相关性及其在膀胱癌不同分期的表达情况,评估KPNB1在膀胱癌中的诊断及预后判断价值。通过细胞实验对KPNB1 在膀胱癌中的作用机制进行初步探索。

结果

在TCGA、GEO 和THPA 数据库中,KPNB1 mRNA 和蛋白水平在膀胱癌组织中明显高于正常膀胱组织。KPNB1 与T 分期、N 分期及肿瘤病理分级显著相关。KPNB1 的表达水平可以有效区分膀胱癌组织与正常膀胱组织(AUC=0.758)。KPNB1 表达较高患者具有较差的无进展生存。KPNB1 过表达组T24 细胞增殖水平以及迁移能力显著提高(P<0.05),KPNB1 敲低组T24 细胞增殖水平以及迁移能力显著下降(P<0.05)。KPNB1 敲低组T24 肿瘤细胞表达STAT3/LDHA 量相对减少(P<0.05),KPNB1 过表达膀胱癌细胞内STAT3/LDHA 表达量显著增加(P<0.05)。STAT3 抑制剂Stattic 处理KPNB1 过表达组膀胱癌细胞后,细胞内LDHA 表达量显著下降(P<0.05)。

结论

KPNB1 在膀胱癌中高表达,且具有较高的诊断及预后判断价值,并通过参与STAT3/LDHA 信号通路在膀胱癌中发挥作用。

Objective

To investigate the expression and role of Karyopherin β1 (KPNB1) in bladder cancer based on bioinformatics analysis and cell studies.

Methods

KPNB1 mRNA and protein levels in normal prostate tissues and prostate cancer tissues were obtained from the Cancer Genome Atlas(TCGA), Gene Expression Omnibus (GEO), and the Human Protein Atlas (THPA) databases. The correlation between KPNB1 expression and clinicopathological features of bladder cancer patients, and the expression of KPNB1 in different subgroups of bladder cancer tissues were analyzed. The diagnostic and prognostic value of KPNB1 in bladder cancer was evaluated. Combined with GOKEEG signal pathway enrichment analysis, the mechanism of KPNB1 in bladder cancer was preliminarily explored. Cellular studies were performed using T24 cells with KPNB1 knockdown or overexpression.

Results

Statistical analysis based on the TGGA, GEO, and THPA databases showed that, compared with normal bladder tissues, KPNB1 mRNA and protein levels were increased in bladder cancer tissues and significantly correlated with N stage, T stage,and degree of tumor invasion. The expression of KPNB1 could effectively distinguish bladder cancer tissues from normal tissues (area under the curve=0.758). KPNB1 expression was higher in patients with poor progression-free survival. The proliferation and migration of T24 cells were significantly increased in the KPNB1 overexpressing group (P<0.05), and the proliferation and migration of T24 cells were significantly decreased in the KPNB1 knockdown group (P<0.05). The expression of STAT3/LDHA in T24 cells was relatively reduced in the KPNB1 knockdown group (P<0.05), and the expression of STAT3/LDHA in KPNB1 overexpressing T24 cells was significantly increased (P<0.05). After treating T24 cells in the KPNB1 overexpressing group with the STAT3 inhibitor Stattic, the expression of LDHA significantly decreased(P<0.05).

Conclusion

KPNB1 is significantly highly expressed in bladder cancer, which is of great diagnostic and prognostic value. KPNB1 plays a role in bladder cancer by participating in the STAT3/LDHA signaling pathway.

表1 膀胱癌患者临床信息分组情况表
图1 KPNB1 mRNA 在膀胱癌组织(n=408)及正常膀胱组织中表达情况(n=19)。图a 为数据库中427 例患者KPNB1mRNA 表达情况;图b 为数据库中19 对膀胱癌与正常配对组织KPNB1 mRNA 表达情况
图2 KPNB1 蛋白在膀胱癌组织及正常膀胱组织中表达情况。图a 为KPNB1 蛋白在正常膀胱组织中的表达情况(抗体CAB03449);图b 为KPNB1 蛋白在膀胱癌组织中的表达情况(抗体 CAB03449);图c 为KPNB1 蛋白在正常膀胱组织中的表达情况(抗体HPA029878);图d 为KPNB1 蛋白在膀胱癌组织的表达情况(抗体HPA029878)
图3 KPNB1 mRNA 在膀胱癌不同亚组中表达情况。图a 为KPNB1 mRNA 在不同T 分期中表达,n=408;图b 为KPNB1 mRNA 在不同N 分期中表达(n=398);图c 为HNRNPA2B1 mRNA 在不同M 分期中表达(n=393);图d 为KPNB1 mRNA 在不同肿瘤病理分级中表达(n=391);图e 为KPNB1 mRNA 在不同年龄中表达(n=408)
表2 KPNB1 表达与419 例膀胱癌患者临床病理特征相关性分析[例(%)]
图4 KPNB1 mRNA 在膀胱癌(n=408)中的诊断价值。图a 为KPNB1mRNA 在膀胱正常组织和膀胱癌组织中的区分价值;图b 为KPNB1mRNA 在膀胱正常组织和T1 期膀胱癌患者组织中的区分价值;图c 为KPNB1 mRNA 在膀胱正常组织和T2~T4 期膀胱癌患者组织中的区分价值
图5 KPNB1mRNA 在膀胱癌(n=408)中的预后价值。图a 为KPNB1mRNA 在膀胱癌中的预后价值;图b 为KPNB1mRNA 在膀胱癌T2~T4 期患者中的预后价值;图c 为KPNB1mRNA 在膀胱癌N0 期患者中的预后价值;图d 为KPNB1mRNA 在N1~N2 期膀胱癌患者中的预后价值;图e 为KPNB1mRNA 在膀胱癌M0 期患者中的预后价值;图f 为KPNB1mRNA 在膀胱癌M1 期患者中的预后价值
图6 核转运蛋白β1(KPNB1)与相关蛋白之间的作用
图7 Western blot 检测T24 细胞中核转运蛋白β1(KPNB1)敲除和过表达后在蛋白水平的表达
图8 噻唑蓝(MTT)实验检测T24 细胞的相对增殖水平
图9 划痕实验检测T24 细胞迁移水平的变化。图a~c 为KPNB1 敲低组、野生型、KPNB1 过表达组T24 细胞0 h 划痕效果图;图d 为KPNB1 敲低组T24 细胞24 h 迁移情况;图e 为KPNB1 野生型T24 细胞24 h 迁移情况;图f 为KPNB1 过表达组T24 细胞24 h 迁移情况
图10 Western blot 检测T24 细胞内STAT3、LDHA信号变化
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