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ADAM8 expression is associated with increased invasiveness and reduced patient survival in pancreatic cancer.

Valkovskaya N, Kayed H, Felix K, Hartmann D, Giese NA, Osinsky SP, Friess H, Kleeff J - J. Cell. Mol. Med. (2007 Sep-Oct)

Bottom Line: ADAM8 mRNA was significantly overexpressed in pancreatic ductal adenocarcinoma (PDAC) compared with normal pancreatic tissues (5.3-fold increase; P= 0.0008), and high ADAM8 mRNA and protein expression levels correlated with reduced survival time of PDAC patients (P= 0.048 and P= 0.065, respectively).In addition, decreased proteolytic activity was measured in cell culture supernatants following silencing of ADAM8.In conclusion, ADAM8 is overexpressed in PDAC, influences cancer cell invasiveness and correlates with reduced survival, suggesting that ADAM8 might be a potential target in pancreatic cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, University of Heidelberg, Heidelberg, Germany.

ABSTRACT
ADAM8 belongs to a family of transmembrane proteins implicated in cell-cell interactions, proteolysis of membrane proteins, and various aspects of carcinogenesis. In the present study, we aimed to evaluate the expression and function of ADAM8 in pancreatic cancer. ADAM8 mRNA levels were analysed by quantitative RT-PCR and correlated to patient survival. Immunohistochemistry was performed to localize ADAM8 in pancreatic tis-sues. Silencing of ADAM8 expression was carried out by transfection with specific siRNA oligonucleotides. Cell growth and invasion assays were used to assess the functional consequences of ADAM8 silencing. SELDI-TOF-MS was performed to detect the proteolytic activity of ADAM8 in pancreatic cancer cells. ADAM8 mRNA was significantly overexpressed in pancreatic ductal adenocarcinoma (PDAC) compared with normal pancreatic tissues (5.3-fold increase; P= 0.0008), and high ADAM8 mRNA and protein expression levels correlated with reduced survival time of PDAC patients (P= 0.048 and P= 0.065, respectively). Silencing of ADAM8 expression did not significantly influence pancreatic cancer cell growth but suppressed invasiveness. In addition, decreased proteolytic activity was measured in cell culture supernatants following silencing of ADAM8. In conclusion, ADAM8 is overexpressed in PDAC, influences cancer cell invasiveness and correlates with reduced survival, suggesting that ADAM8 might be a potential target in pancreatic cancer therapy.

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ADAM8 expression and localization in the normal pancreas, CP and PDAC tissues: Immunohistochemistry was performed as described in the ‘Materials and methods’ section. (A) ADAM8 expression in normal pancreatic islets (arrows) and acinar cells and ductal cells (lower inset). Consecutive section stained with CK19 confirming ductal cells (upper inset). (B) ADAM8 expression in CP tissues (arrows indicate tubular complexes). Consecutive section stained with CK19 confirming ductal cells/tubular complexes (inset). (C, D) ADAM8 (C) and CK19 (D) staining in pancreatic cancer cells of PDAC tissues. (E, F) Strong ADAM8 staining in pancreatic cancer cells of PDAC tissues. Note the absent staining in a consecutive negative control tissue section (inset). Horizontal lines represent the scale bar of 50 μm. (G) Survival curves of PDAC patients (n = 99) with weak/moderate ADAM8 staining versus strong ADAM8 staining (P= 0.065).
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fig03: ADAM8 expression and localization in the normal pancreas, CP and PDAC tissues: Immunohistochemistry was performed as described in the ‘Materials and methods’ section. (A) ADAM8 expression in normal pancreatic islets (arrows) and acinar cells and ductal cells (lower inset). Consecutive section stained with CK19 confirming ductal cells (upper inset). (B) ADAM8 expression in CP tissues (arrows indicate tubular complexes). Consecutive section stained with CK19 confirming ductal cells/tubular complexes (inset). (C, D) ADAM8 (C) and CK19 (D) staining in pancreatic cancer cells of PDAC tissues. (E, F) Strong ADAM8 staining in pancreatic cancer cells of PDAC tissues. Note the absent staining in a consecutive negative control tissue section (inset). Horizontal lines represent the scale bar of 50 μm. (G) Survival curves of PDAC patients (n = 99) with weak/moderate ADAM8 staining versus strong ADAM8 staining (P= 0.065).

Mentions: In order to determine the exact localization of ADAM8 in CP and PDAC, immunohistochemistry was carried out next. In normal pancreatic tissues (n = 8), ADAM8 exhibited weak cytoplasmic and membranous staining in normal ductal and acinar cells (Fig. 3A, lower inset) and moderate expression in islet cells (Fig. 3A). Ductal cells were confirmed by consecutive sections stained with CK19 as a ductal cell marker (Fig. 3A, upper inset; 3B, inset). ADAM8 expression was also weakly observed in the stroma, nerves and blood vessels of normal pancreatic tis-sues. In CP tissues (n = 8), ADAM8 exhibited moderate to strong staining in ductal cells, tubular complexes and degenerating acinar cells (Fig. 3B) but not in inflammatory cells. In PDAC tissues (n = 99), ADAM8 demonstrated moderate-to-strong staining on the plasma membrane of the cancer cells in 79% of the cases (Fig. 3C, E and F). Pancreatic ductal adeno-carcinoma cells were also positive for CK19 in consecutive sections (Fig. 3D). In addition, ADAM8 expression was detected in tubular complexes of 36% of the cases, and in degenerating acinar cells of 51% of the cases. Weak ADAM8 staining was observed in intra-pancreatic nerves. In lymph node metastases (n = 6) of PDAC, ADAM8 exhibited moderate to strong staining in the metastatic cancer cells of all cases. The specificity of ADAM8 staining was demonstrated by the absent staining in consecutive negative control tissue sections (Fig. 3F, inset). Next, a semi-quantification analysis for the intensity of ADAM8 staining in pancreatic cancer cells in PDAC tissues was performed. Staining was categorized as absent (n = 0), weak (n = 21), moderate (n = 46), and strong (n = 32). There was a tendency towards better survival of PDAC patients exhibiting weak/moderate ADAM8 staining (median survival: 20 months), in comparison to patients exhibiting strong ADAM8 staining in pancreatic cancer cells (median survival: 14 months; P= 0.065) (Fig. 3G).


ADAM8 expression is associated with increased invasiveness and reduced patient survival in pancreatic cancer.

Valkovskaya N, Kayed H, Felix K, Hartmann D, Giese NA, Osinsky SP, Friess H, Kleeff J - J. Cell. Mol. Med. (2007 Sep-Oct)

ADAM8 expression and localization in the normal pancreas, CP and PDAC tissues: Immunohistochemistry was performed as described in the ‘Materials and methods’ section. (A) ADAM8 expression in normal pancreatic islets (arrows) and acinar cells and ductal cells (lower inset). Consecutive section stained with CK19 confirming ductal cells (upper inset). (B) ADAM8 expression in CP tissues (arrows indicate tubular complexes). Consecutive section stained with CK19 confirming ductal cells/tubular complexes (inset). (C, D) ADAM8 (C) and CK19 (D) staining in pancreatic cancer cells of PDAC tissues. (E, F) Strong ADAM8 staining in pancreatic cancer cells of PDAC tissues. Note the absent staining in a consecutive negative control tissue section (inset). Horizontal lines represent the scale bar of 50 μm. (G) Survival curves of PDAC patients (n = 99) with weak/moderate ADAM8 staining versus strong ADAM8 staining (P= 0.065).
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fig03: ADAM8 expression and localization in the normal pancreas, CP and PDAC tissues: Immunohistochemistry was performed as described in the ‘Materials and methods’ section. (A) ADAM8 expression in normal pancreatic islets (arrows) and acinar cells and ductal cells (lower inset). Consecutive section stained with CK19 confirming ductal cells (upper inset). (B) ADAM8 expression in CP tissues (arrows indicate tubular complexes). Consecutive section stained with CK19 confirming ductal cells/tubular complexes (inset). (C, D) ADAM8 (C) and CK19 (D) staining in pancreatic cancer cells of PDAC tissues. (E, F) Strong ADAM8 staining in pancreatic cancer cells of PDAC tissues. Note the absent staining in a consecutive negative control tissue section (inset). Horizontal lines represent the scale bar of 50 μm. (G) Survival curves of PDAC patients (n = 99) with weak/moderate ADAM8 staining versus strong ADAM8 staining (P= 0.065).
Mentions: In order to determine the exact localization of ADAM8 in CP and PDAC, immunohistochemistry was carried out next. In normal pancreatic tissues (n = 8), ADAM8 exhibited weak cytoplasmic and membranous staining in normal ductal and acinar cells (Fig. 3A, lower inset) and moderate expression in islet cells (Fig. 3A). Ductal cells were confirmed by consecutive sections stained with CK19 as a ductal cell marker (Fig. 3A, upper inset; 3B, inset). ADAM8 expression was also weakly observed in the stroma, nerves and blood vessels of normal pancreatic tis-sues. In CP tissues (n = 8), ADAM8 exhibited moderate to strong staining in ductal cells, tubular complexes and degenerating acinar cells (Fig. 3B) but not in inflammatory cells. In PDAC tissues (n = 99), ADAM8 demonstrated moderate-to-strong staining on the plasma membrane of the cancer cells in 79% of the cases (Fig. 3C, E and F). Pancreatic ductal adeno-carcinoma cells were also positive for CK19 in consecutive sections (Fig. 3D). In addition, ADAM8 expression was detected in tubular complexes of 36% of the cases, and in degenerating acinar cells of 51% of the cases. Weak ADAM8 staining was observed in intra-pancreatic nerves. In lymph node metastases (n = 6) of PDAC, ADAM8 exhibited moderate to strong staining in the metastatic cancer cells of all cases. The specificity of ADAM8 staining was demonstrated by the absent staining in consecutive negative control tissue sections (Fig. 3F, inset). Next, a semi-quantification analysis for the intensity of ADAM8 staining in pancreatic cancer cells in PDAC tissues was performed. Staining was categorized as absent (n = 0), weak (n = 21), moderate (n = 46), and strong (n = 32). There was a tendency towards better survival of PDAC patients exhibiting weak/moderate ADAM8 staining (median survival: 20 months), in comparison to patients exhibiting strong ADAM8 staining in pancreatic cancer cells (median survival: 14 months; P= 0.065) (Fig. 3G).

Bottom Line: ADAM8 mRNA was significantly overexpressed in pancreatic ductal adenocarcinoma (PDAC) compared with normal pancreatic tissues (5.3-fold increase; P= 0.0008), and high ADAM8 mRNA and protein expression levels correlated with reduced survival time of PDAC patients (P= 0.048 and P= 0.065, respectively).In addition, decreased proteolytic activity was measured in cell culture supernatants following silencing of ADAM8.In conclusion, ADAM8 is overexpressed in PDAC, influences cancer cell invasiveness and correlates with reduced survival, suggesting that ADAM8 might be a potential target in pancreatic cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, University of Heidelberg, Heidelberg, Germany.

ABSTRACT
ADAM8 belongs to a family of transmembrane proteins implicated in cell-cell interactions, proteolysis of membrane proteins, and various aspects of carcinogenesis. In the present study, we aimed to evaluate the expression and function of ADAM8 in pancreatic cancer. ADAM8 mRNA levels were analysed by quantitative RT-PCR and correlated to patient survival. Immunohistochemistry was performed to localize ADAM8 in pancreatic tis-sues. Silencing of ADAM8 expression was carried out by transfection with specific siRNA oligonucleotides. Cell growth and invasion assays were used to assess the functional consequences of ADAM8 silencing. SELDI-TOF-MS was performed to detect the proteolytic activity of ADAM8 in pancreatic cancer cells. ADAM8 mRNA was significantly overexpressed in pancreatic ductal adenocarcinoma (PDAC) compared with normal pancreatic tissues (5.3-fold increase; P= 0.0008), and high ADAM8 mRNA and protein expression levels correlated with reduced survival time of PDAC patients (P= 0.048 and P= 0.065, respectively). Silencing of ADAM8 expression did not significantly influence pancreatic cancer cell growth but suppressed invasiveness. In addition, decreased proteolytic activity was measured in cell culture supernatants following silencing of ADAM8. In conclusion, ADAM8 is overexpressed in PDAC, influences cancer cell invasiveness and correlates with reduced survival, suggesting that ADAM8 might be a potential target in pancreatic cancer therapy.

Show MeSH
Related in: MedlinePlus