Comprehensive profiling and localisation of the matrix metalloproteinases in urothelial carcinoma

Abstract

The matrix metalloproteinases (MMPs) are endopeptidases which break down the extracellular matrix and regulate cytokine and growth factor activity. Several MMPs have been implicated in the promotion of invasion and metastasis in a broad range of tumours including urothelial carcinoma. In this study, RNA from 132 normal bladder and urothelial carcinoma specimens was profiled for each of the 24 human MMPs, the four endogenous tissue inhibitors of MMPs (TIMPs) and several key growth factors and their receptors using quantitative real time RT -PCR. Laser capture microdissection (LCM) of RNA from 22 tumour and 11 normal frozen sections was performed allowing accurate RNA extraction from either stromal or epithelial compartments. This study confirms the over expression in bladder tumour tissue of well-documented MMPs and highlights a range of MMPs which have not previously been implicated in the development of urothelial cancer. In summary, MMP-2, MT1-MMP and the previously unreported MMP-28 were very highly expressed in tumour samples while MMPs 1, 7, 9, 11, 15, 19 and 23 were highly expressed. There was a significant positive correlation between transcript expression and tumour grade for MMPs 1, 2, 8, 10, 11, 12, 13, 14, 15 and 28 (Po0.001). At the same confidence interval, TIMP-1 and TIMP-3 also correlated with increasing tumour grade. LCM revealed that most highly expressed MMPs are located primarily within the stromal compartment except MMP-13 which localised to the epithelial compartment. This work forms the basis for further functional studies, which will help to confirm the MMPs as potential diagnostic and therapeutic targets in early bladder cancer.

Human matrix metalloproteinases: An ubiquitarian class of enzymes involved in several pathological processes

“…Increased expression of MMP-28 was observed in a variety of tumors and tumor cell lines and its role seems to vary based on tumor type and stage of the disease Wallard et al, 2006). Only in the case of colon carcinoma MMP-28, unlike other MMPs, is downregulated, as compared to normal tissues (Bister et al, 2004).…”

“…Neoplastic diseases. Elevated MT6-MMP mRNA expression was found in several human cancers including brain (anaplastic astrocytomas and glioblastomas) Velasco et al, 2000), colon (Velasco et al, 2000), urothelial (Wallard et al, 2006), and prostate (Riddick et al, 2005) cancers. In gliomas, expression of MT6-MMP mRNA was suggested to contribute to the disease progression .…”

UHRF1 is a novel molecular marker for diagnosis and the prognosis of bladder cancer

“…RNA was extracted using an RNeasy Micro Kit (Qiagen, Crawley, UK). Areas of cancer or stroma containing significant inflammatory cell infiltration were avoided to prevent contamination (Wallard et al, 2006). Total RNA was treated with DNase and then quantitative reverse transcription -PCR (qRT -PCR) was performed as described below.…”

Neural MMP‐28 expression precedes myelination during development and peripheral nerve repair

“…As is common with other MMPs, MMP-28 has been shown to be expressed at increased levels in multiple disease states (Overall and Kleifeld, 2006). In some tumors and cancer cell lines, an increase in MMP-28 was detected (Saarialho-Kere et al, 2002;Lin et al, 2006;Wallard et al, 2006), although in some cases, MMP-28 protein may be lower in cancer compared with normal tissue (Bister et al, 2004). In wounded skin, strong up-regulation of MMP-28 occurs in mitotic cells behind the advancing wound edge, but not in the actively migrating keratinocytes that secrete various other MMPs such as collagenase, stromelysin, and gelatinase (Saarialho-Kere et al, 2002).…”