南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (8): 1109-1118.doi: 10.12122/j.issn.1673-4254.2022.08.01

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MYBL2在前列腺癌患者组织中高表达并与不良预后相关

杨 明，朱旭东，沈 炀，何 麒，秦 远，邵轶群，袁 琳，叶和松   

  1. 南京中医药大学第二附属医院泌尿外科，江苏 南京 210017；上海中医药大学附属岳阳中西医结合医院泌尿外科，上海 200437；南京中医药大学附属医院泌尿外科，江苏 南京 210029
  • 出版日期:2022-08-20 发布日期:2022-09-05

High expression of MYBL2 promotes progression and predicts a poor survival outcome of prostate cancer

YANG Ming, ZHU Xudong, SHEN Yang, HE Qi, QIN Yuan, SHAO Yiqun, YUAN Lin, YE Hesong   

  1. Department of Urology, The Second Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210017, China; Department of Urology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China; Department of Urology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
  • Online:2022-08-20 Published:2022-09-05

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摘要： 目的 探索骨髓母细胞增生症病毒癌基因同源物样2（MYBL2）基因对前列腺癌（PCa）患者临床预后和生物学行为的影响。方法 采用qRT-PCR检测MYBL2在45例PCa与癌旁前列腺组织的表达水平，根据MYBL2表达中位数，分为高（23例）低（22例）表达，采用非参数检验、Kaplan-Meier、单因素和多因素Cox法，并分析MYBL2高低表达与PCa的临床病理特征及预后相关性，利用癌症基因组图谱（TCGA）基因芯片数据库PCa数据集进行验证。基因集富集分析（GSEA）MYBL2高低表达可能参与调控的分子通路，CIBERSORT算法研究MYBL2与肿瘤免疫微环境的相关性。最后，体内实验中，通过建立阴性对照组（shCtrl）、敲减MYBL2组（sh-MYBL2）用细胞增殖毒性检测试剂盒（CCK8）和Transwell法检测各组细胞增殖和侵袭能力。qRT-PCR和Western blotting检测肿瘤细胞中MYBL2基因和蛋白的表达。在体内实验中，检测2组荷瘤小鼠瘤重和免疫组织化学方法观察荷瘤组织Ki-67的表达水平。结果 两组数据均显示：MYBL2在PCa组织中高表达，与Gleason评分、临床和病理分期正相关（P<0.01），与年龄无相关性；Kaplan-Meier分析MYBL2高表达与无生化复发生存期显著负相关（P<0.05），与总生存期无关。Cox回归分析：TCGA数据集中临床和病理分期，我们的数据中临床分期和Gleason评分是PCa无复发生存期的独立预后因素（P<0.05）。GSEA结果显示高表达MYBL2与免疫、细胞粘附、细胞因子等通路有关。CIBERSORT分析：MYBL2表达与B cells memory和Mast cells resting有关（P<0.05）。体外研究显示，与shCtrl组相比，shMYBL2组能显著抑制LNCaP、PC-3细胞增殖和侵袭（P<0.01）。体内研究显示，shMYBL2组PC-3荷瘤小鼠体内肿瘤的平均重和Ki67阳性表达率显著低于shCtrl组（P<0.01）。结论 MYBL2是一种致癌基因，与PCa多个病理指标相关，可以作为潜在的诊疗PCa患者预后的标志物和治疗肿瘤的靶标。

关键词: MYBL2；前列腺癌；细胞增殖；细胞侵袭；荷瘤小鼠；预后

Abstract: Objective To explore the correlation of MYB proto-oncogene like 2 (MYBL2) with biological behaviors and clinical prognosis of prostate cancer (PCa). Methods We detected Mybl2 mRNA expression in 45 pairs of PCa and adjacent tissues using real-time quantitative PCR, and analyzed the correlation of high (23 cases) and low expression (22 cases) of Mybl2 with clinicopathological features and prognosis of the patients using nonparametric test, Kaplan-Meier survival analysis and univariate and multivariate Cox regression. The results were verified by analysis of the data from Cancer Genome Atlas (TCGA) microarray database, and the molecular pathways were identified by gene set enrichment analysis (GSEA). The CIBERPORT algorithm was used to identify the correlations between Mybl2 expression and tumor microenvironment of PCa. We also tested the effects of MYBL2 knockdown on proliferation and invasion of PCa cell lines using cell counting kit-8 and Transwell assays and observed the growth of PC3 cell xenograft with MYBL2 knockdown in nude mice and the expression levels of Ki-67 in the xenograft using immunohistochemistry. Results Mybl2 expression was significantly elevated in PCa tissues in close correlation with Gleason score and clinical and pathological stage of the tumor (P<0.01) but not with the patients' age. Kaplan-Meier analysis indicated a significant negative correlation of high Mybl2 expression with recurrence-free survival (P<0.05), but not with the overall survival of the patients. The data from TCGA suggested that clinical and pathological stages were independent prognostic factors for recurrence-free survival, and our data indicated that clinical stage and Gleason score were independent prognostic factors of PCa (P<0.05). GSEA suggested that Mybl2 expression was related with the pathways involving immune function, cell adhesion, and cytokine secretion; CIBERPORT analysis suggested the involvement of Mybl2 expression with memory B cells and resting mast cells (P<0.05). In LNCaP and PC-3 cells, MYBL2 knockdown significantly inhibited cell proliferation and invasion (P<0.05); in the tumor-bearing nude mice, the xenografts derived from PC-3 cells with MYBL2 knockdown exhibited a lowered mean tumor weight and positivity rate for Ki67 (P<0.05). Conclusion Mybl2 is an oncogene related with multiple pathological indicators of PCa and can serve as a potential prognostic marker as well as a therapeutic target for patients with PCa.

Key words: Mybl2; prostate cancer; cell proliferation; cell invasion; tumor-bearing mice; prognosis