Inhibition of MMP-9 using a pyrrole-imidazole polyamide reduces cell invasion in renal cell carcinoma

Abstract

Abstract.We investigated the clinical significance of the expression levels of matrix metalloproteinase 9 (MMP-9) in renal cell carcinoma (RCC). In addition, we validated the efficacy of pyrrole imidazole polyamide (PIP) targeting MMP-9 on inhibiting proliferation and invasion of RCC. We evaluated the expression levels of MMP-9 in 249 RCC specimens by immunostaining and analyzed the association between MMP-9 expression levels and cancer-specific survival. Furthermore, in a human RCC cell line, Caki-2, we tested the effect of a couple of PIPs targeting MMP-9 one recognizing an NF-κB binding site (MMP-9-NF-κB PIP) and another for the AP-1 binding site (MMP-9-AP-1 PIP) in the MMP-9 promoter. The expression levels of MMP-9, proliferative activity and invasive capability were tested by quantitative PCR, WST8 assay and Matrigel invasion assay, respectively. By immunostaining of the clinical specimens, strong MMP-9 staining was proven to be a significant predictor of poor prognosis for cancer-specific survival (P<0.01). In Caki-2 cells, MMP-9-NF-κB PIP significantly reduced the expression levels of MMP-9 mRNA and inhibited cell invasion, but did not affect the cell proliferation activity. On the other hand, no effect was found in MMP-9-AP-1 PIP on MMP-9 mRNA expression, cell proliferation and invasion. We confirmed the inhibitory effects of MMP-9-NF-κB PIP on the expression of MMP-9 and subsequent invasion of Caki-2 cells. Since it was clearly shown that high MMP-9 expression levels were associated with poor prognosis of RCC, MMP-9 is a potential candidate target for RCC treatment. Transcription therapy using a minor groove binder, such as NF-κB PIP, may be a potential therapeutic agent for RCC, although further investigation is required.

Melatonin inhibits MMP‐9 transactivation and renal cell carcinoma metastasis by suppressing Akt‐MAPKs pathway and NF‐κB DNA‐binding activity

“…A previous report indicated that substantially higher MMP‐9 protein expression was observed in RCC tumors than in normal tissues, but no difference was observed in MMP‐2 expression . MMP‐9 could be a critical molecule for RCC invasion and metastasis not only because this MMP type destroys type IV collagen, a major component of the basement membrane , but also because type IV collagenase activity is a key determinant of the in vitro invasiveness of RCC cells and their metastatic potential . Our results are consistent with those of previous studies, which have indicated that MMP‐9 is a critical target of melatonin for regulation of RCC metastasis.…”

“…The invasion of the basement membrane proceeds through a series of discrete steps, and the degradation of matrix in the basement membrane is largely controlled by activities of various matrix metalloproteinase (MMP) subtypes . MMPs are overexpressed in almost all cancers including RCC and are believed to be a promising therapeutic target for RCC . Among these MMPs, MMP‐2, MMP‐9, and their endogenous inhibitors, namely tissue inhibitor of matrix metalloproteinase (TIMP)‐1 and TIMP‐2, are the most crucial enzymes for controlling the degradation of the main constituent of the extracellular matrix (ECM), type IV collagen, and are deeply involved in cancer invasion and metastasis .…”

New miRNA Profiles Accurately Distinguish Renal Cell Carcinomas and Upper Tract Urothelial Carcinomas from the Normal Kidney

“…Furthermore, the systemic inflammatory response has been revealed to contribute to tumorigenesis in the last two decades. Key inflammatory pathways, such as the NF-κB [45], AP-1 [46] and STAT3 [47] pathways, are implicated in the proliferation, transformation, survival, invasion, angiogenesis, metastasis, chemoresistance and radioresistance of cancer [48]. Systemic inflammatory response, represented by elevated C-reactive protein (CRP), platelet count (PC) and erythrocyte sedimentation rate (ESR) was recently shown to predict poor survival in patients with RCC [49].…”

Alpha-Mangostin Suppresses the Metastasis of Human Renal Carcinoma Cells by Targeting MEK/ERK Expression and MMP-9 Transcription Activity

“…Other studies reported that increased phosphorylation of ERK and enhanced expression of MMP-2 or MMP-9 by treatment with gamma-aminobutyric acid (an inhibitory neurotransmitter) or knockdown of hepatocellular carcinoma-related protein (HCRP) increased the invasive activity of RCC cells [30]. Additional evidence indicates that a pyrrole-imidazole polyamide specifically targets binding sites on the MMP-9 promoter, and thereby inhibits the invasion (but not proliferation) of RCC cells [14]. We demonstrated that α-mangostin had inhibitory effects on the MMP-9 transcription activity and protein expression.…”

“…The matrix metalloproteinases (MMPs), crucial proteolytic proteinases that allow malignant cells to access the vasculature, are a family of zinc-dependent endopeptidases with broad substrate specificities for a variety of extracellular matrix (ECM) and basement membrane proteins [14]. Aberrant regulation of the MMPs disrupts normal cellular interactions, and leads to lysis of the ECM and breaching of the basement membrane, thereby allowing malignant cells to migrate and invade distant tissues [15].…”