Singh R, Mandhani A, Agrawal V, Garg Minal
Department of Biochemistry, Lucknow University, Lucknow, 226007, India.
Uro-Oncology and Minimally Invasive Surgery, Fortis Escorts Kidney & Urology, New Delhi, India.
Cancer Microenviron. 2018 Jun;11(1):23-39. doi: 10.1007/s12307-017-0199-4. Epub 2018 Jan 18.
Involvement of matrix metalloproteinases (MMPs) in the pathogenesis of urothelial carcinoma elects them to be sensitive marker for clinical and prognostic implications. MMPs regulate tumor growth and invasion by inducing epithelial-to-mesenchymal transition (EMT) which is characterized by the complex reprogramming of epithelial cells and ultimately bring about major changes in the structural organization of bladder urothelium. The present study has been undertaken to evaluate the clinical relevance of MMPs in two distinct types of bladder cancer disease. Expression analysis of MMPs namely MMP-2, MMP-7, MMP-9 and EMT markers including epithelial marker, E-cadherin; mesenchymal markers, N-cadherin and Vimentin; and EMT-activating transcriptional factors (EMT-ATFs), Snail, Slug, Twist and Zeb was done in 64 cases of bladder tumor tissues [{Non-muscle invasive bladder cancer (NMIBC): 35 cases} and {Muscle invasive bladder cancer (MIBC): 29 cases}] by real-time quantitative polymerase chain reaction (RT-qPCR). Immunohistochemistry (IHC) staining was done in matched bladder tumor tissues to evaluate the protein expression and localization of E-cadherin, N-cadherin, Vimentin, Snail, and Slug. Our data showed overexpression of MMP-2, MMP-7 and MMP-9 at transcriptome level in 32.8%, 25% and 37.5% bladder tumor cases respectively. These tumor tissues were examined for higher expression of mesenchymal markers (N-cadherin and Vimentin) at mRNA and protein level and exhibited statistical association with tumor stage and tumor grade (p = 0.02, p = 0.04, Mann-Whitney test). Significant statistical correlation in tumor tissues with overexpressed MMPs has also been observed between gain of transcriptional factors and weak expression of E-cadherin with tumor stage, grade, gender, presence of hematuria and smoking history of the patients. Gene expression patterns of EMT markers in bladder tumors with overexpressed MMPs and their significant association with clinical profile validate the important role of MMPs in the pathogenesis of urothelial carcinoma of bladder (UCB). Increased expression of specific MMPs may affect several downstream EMT programs and thus may improve its diagnostic and prognostic utility in clinical setting.
基质金属蛋白酶（MMPs）参与尿路上皮癌的发病机制，使其成为具有临床和预后意义的敏感标志物。MMPs通过诱导上皮-间质转化（EMT）来调节肿瘤生长和侵袭，EMT的特征是上皮细胞的复杂重编程，最终导致膀胱尿路上皮结构组织的重大变化。本研究旨在评估MMPs在两种不同类型膀胱癌疾病中的临床相关性。通过实时定量聚合酶链反应（RT-qPCR）对64例膀胱肿瘤组织[非肌层浸润性膀胱癌（NMIBC）：35例和肌层浸润性膀胱癌（MIBC）：29例]进行MMP-2、MMP-7、MMP-9等MMPs以及包括上皮标志物E-钙黏蛋白、间质标志物N-钙黏蛋白和波形蛋白，以及EMT激活转录因子（EMT-ATFs）蜗牛蛋白、蛞蝓蛋白、Twist和锌指蛋白E盒结合因子（Zeb）的表达分析。对匹配的膀胱肿瘤组织进行免疫组织化学（IHC）染色，以评估E-钙黏蛋白、N-钙黏蛋白、波形蛋白、蜗牛蛋白和蛞蝓蛋白的蛋白表达和定位。我们的数据显示，在转录组水平上，MMP-2、MMP-7和MMP-9在膀胱肿瘤病例中的过表达率分别为32.8%、25%和37.5%。对这些肿瘤组织进行检测，发现间质标志物（N-钙黏蛋白和波形蛋白）在mRNA和蛋白水平上表达较高，且与肿瘤分期和肿瘤分级具有统计学关联（p = 0.02，p = 0.04，曼-惠特尼检验）。在过表达MMPs的肿瘤组织中，还观察到转录因子的增加与E-钙黏蛋白的弱表达与患者的肿瘤分期、分级、性别、血尿情况和吸烟史之间存在显著的统计学相关性。MMPs过表达的膀胱肿瘤中EMT标志物的基因表达模式及其与临床特征的显著关联，证实了MMPs在膀胱尿路上皮癌（UCB）发病机制中的重要作用。特定MMPs表达的增加可能会影响多个下游EMT程序，从而可能提高其在临床环境中的诊断和预后效用。
