MMP, VEGF and TIMP as prognostic factors in recurring bladder cancer

doi:10.1016/j.clinbiochem.2015.07.021

Clinical Biochemistry

Volume 48, Issue 18, December 2015, Pages 1235-1240

https://doi.org/10.1016/j.clinbiochem.2015.07.021 Get rights and content

Highlights

•
We investigate the clinical correlates and prognostic utility of MMP, VEGF and TIMP genes expression in BCa recurrence.
•
Expression of MMP9 gene in PBLs of patients at diagnosis is associated with the differentiation grade, and smoking status.
•
Expression of MMP9, VEGFA and TIMP1 genes in PBLs may play a pivotal role in regulation of progression of BCa.
•
Expression of TIMP1 gene may be important factor for early recurrence of BCa.

Abstract

Objectives

To investigate the clinical correlates and prognostic utility of MMP, VEGF and TIMP genes expression in bladder cancer (BCa) recurrence.

Methods

Expression of MMP1, MMP2, MMP9, VEGFA and TIMP1, TIMP3 was analyzed by qRT-PCR using SYBR Green in peripheral blood leukocytes (PBLs) of BCa patients at two time points (diagnosis (n = 40), and first recurrence (n = 40)) and an age-matched group of healthy controls (n = 100). Plasma concentrations of MMP1 (pro- and active forms) were measured using ELISA in BCa patients.

Results

The expression of MMP1 mRNA was significantly lower in BCa patients with first recurrence compared to control (p = 0.019). Expression of other genes did not differ significantly between the groups. MMP9 gene expression was associated with differentiation grade (p = 0.043), with the highest expression in poorly differentiated tumors (G3) and was higher in smokers than in non-smokers (p = 0.039) in BCa patients at diagnosis. The results at two time points showed that MMP9 and VEGFA genes expression was increased in patients with moderately differentiated BCa (p = 0.029), and advanced pathologic stage (p = 0.048), respectively. Moreover, gene expression of TIMP1 was increased for G3 (p = 0.043), and was decreased for early recurrence (p = 0.003).

Conclusions

Our study suggests that the expression of MMP9 in PBLs of BCa patients at diagnosis is associated with the differentiation grade of the BCa, and smoking status. Genes expression of MMP9, VEGFA and TIMP1 in PBLs may play a pivotal role in regulation of progression of BCa. Additionally, TIMP1 gene expression may be important factor for early recurrence of BCa.

Graphical abstract

Introduction

Transurethral resection of BCa (TURB) is standard treatment associated with NMIBC's residual risk. While partial cystectomy is a bladder sparing procedure, it is applied to treat invasive bladder cancer in selected patients [1]. More invasive treatment including combination of TURB with radiotherapy or chemotherapy, which may improve the recurrence-free survival, is of limited applicability, because of the patients' age-related health conditions [2]. Therefore, effective surveillance of BCa and development of efficient predictors of its recurrence are still necessary.

Tumor invasion depends on the remodeling processes that occur in connective tissue extracellular matrix (ECM). Matrix metalloproteinases (MMPs) are the most important extracellular proteases that play a role in degradation and dissolution of ECM and basement membrane [3], [4]. MMPs cooperate in the regulation of proangiogenic factor — vascular endothelial growth factor (VEGF) [5], [6]. The proteolytic activity of MMP enzymes is highly regulated by effective inhibitors — tissue inhibitors of metalloproteinases (TIMPs) [7].

The basement membrane damage and angiogenesis processes involve interactions between various ECM proteins, in which MMPs, VEGFs and TIMPs seem to be most important [8], [9], [10]. Several reports have demonstrated that the expression of MMP, TIMP and VEGF genes is correlated with tumor progression, invasion and angiogenesis of BCa [11], [12], [13], [14], [15], [16]. Despite the evidence of involvement of the MMPs, TIMPs and VEGFs in tumorigenesis and progression of BCa, little is known about their practical usefulness as non-invasive prognostic tools for predicting early recurrence after treatment of BCa. Therefore, we have chosen to investigate whether expression of MMP, TIMP, VEGF genes in peripheral blood leukocytes (PBLs) of BCa patients correlates with clinicopathological features and recurrence. Moreover, we have evaluated the importance of plasma MMP1 levels at diagnosis for prediction of recurrence.

Section snippets

Patients and controls

BCa patients and controls were recruited from the First Department of Urology, Medical University of Lodz and Nofer Institute of Occupational Medicine (NIOM) (Lodz, Poland). Blood samples from BCa patients were collected between 2005–2011. From this group, we selected forty patients with TURB or partial cystectomy and with recurrence. A total of the 100 age-matched controls were recruited between 2011–2012 (Fig. 1). None of the BCa patients showed evidence of any other malignancies at the time

Subject characteristics

One hundred forty individuals (40 BCa patients and 100 controls) were included in the study. Baseline clinical characteristics of BCa patients at diagnosis and controls are shown in Supplementary Table 2. There were no statistically significant differences between the BCa patients and controls for sex and age distribution (p = 0.815 and p = 0.151, respectively). Comparison of body mass index (BMI), indicated that controls were more obese than BCa patients at diagnosis (p = 0.001). There were

Discussion

In this study we analyzed expression of MMP1, MMP2, MMP9, VEGFA and TIMP1, TIMP3 genes in PBLs as potential prognostic factors for recurrence after TURB or partial cystectomy in BCa patients. We also attempted to determine the plasma MMP1 levels and clinical characteristics of BCa in relation to recurrence. To the best of our knowledge, the use of the expression of these genes for the evaluation of recurrence in BCa patients after TURB or partial cystectomy has not been previously performed.

Conflict of interest

None declared.

Acknowledgments

This work was supported by Nofer Institute of Occupational Medicine (internal grant No. 1.22/2013) and the Polish Society of Urology (No. 01/2012).

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View full text

MMP, VEGF and TIMP as prognostic factors in recurring bladder cancer

Highlights

Abstract

Objectives

Methods

Results

Conclusions

Graphical abstract

Introduction

Section snippets

Patients and controls

Subject characteristics

Discussion

Conflict of interest

Acknowledgments

J. Urol.

Actas Urol. Esp.

Eur. Urol.

Matrix Biol.

Clin. Chim. Acta

J. Lab. Clin. Med.

Eur. J. Cancer

J. Urol.

Urologic Oncology: Seminars and Original Investigations

Actas Urol. Esp.

J. Urol.

Age and comorbidity impact surgical therapy in older bladder carcinoma patients: a population-based study

Cancer

New and paradoxical roles of matrix metalloproteinases in the tumor microenvironment

Front. Pharmacol.

Matrix metalloproteinases: protective roles in cancer

J. Cell. Mol. Med.

Vegf targets the tumour cell

Nat. Rev. Cancer

Vascular endothelial growth factor and the regulation of angiogenesis

Recent Prog. Horm. Res.

Tissue inhibitors of metalloproteinases

Genome Biol.

Matrix metalloproteinases and tumor metastasis

Cancer Metastasis Rev.

Matrix metalloproteinases and their clinical relevance in urinary bladder cancer

Nat. Rev. Urol.