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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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Related in: MedlinePlus

SELDI-TOF-MS analysis. Proteolytic activity of ADAM8 in cell culture super-natants assessed by SELDI-TOF-MS. (A) Trace-view and gel view of the mass spectra between 30 and 50 kD of the control siRNA and ADAM8 siRNA #1 transfected cells. Arrow points to the 39.2 kD protein and its suppression in siRNA transfected cells. (B) Decreased intensities of protein peak (arrows) in control siRNA transfected cells.
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fig06: SELDI-TOF-MS analysis. Proteolytic activity of ADAM8 in cell culture super-natants assessed by SELDI-TOF-MS. (A) Trace-view and gel view of the mass spectra between 30 and 50 kD of the control siRNA and ADAM8 siRNA #1 transfected cells. Arrow points to the 39.2 kD protein and its suppression in siRNA transfected cells. (B) Decreased intensities of protein peak (arrows) in control siRNA transfected cells.

Mentions: The data were further analysed by Biomarker Wizard Software, a univariate analysis tool. After analysis of the intensity values for all spectra at a given mass, there were significant differences in distribution of proteins in the control siRNA transfected ASPC-1 cell culture supernatants compared to the ADAM8 siRNA transfected ASPC-1 supernatants. It could be shown (Fig. 6A) that the peak at m/z 39,232 was more than three-fold higher in control super-natants (height from baseline 2.8) compared to siRNA supernatants (height from baseline 0.8). The lower intensity of this protein in ASPC-1 ADAM8 siRNA transfected cell culture supernatants leads to the conclusion that it is probably a shedded part of ADAM8. The differential profiling of ADAM8 and control transfected cell supernatant spectra revealed in the controls not only a decreased intensity of some protein peaks but also a complete disappearance of some proteins, as shown in the boxed area covering the 5–7 kD range (Fig. 6B). The presence of additional peaks in supernatants of the ADAM8 siRNA trans-fected cell cultures indicates the knock-down ADAM8 expression, and consequently a lower proteolytic activity in these cell culture supernatants.


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)

SELDI-TOF-MS analysis. Proteolytic activity of ADAM8 in cell culture super-natants assessed by SELDI-TOF-MS. (A) Trace-view and gel view of the mass spectra between 30 and 50 kD of the control siRNA and ADAM8 siRNA #1 transfected cells. Arrow points to the 39.2 kD protein and its suppression in siRNA transfected cells. (B) Decreased intensities of protein peak (arrows) in control siRNA transfected cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401277&req=5

fig06: SELDI-TOF-MS analysis. Proteolytic activity of ADAM8 in cell culture super-natants assessed by SELDI-TOF-MS. (A) Trace-view and gel view of the mass spectra between 30 and 50 kD of the control siRNA and ADAM8 siRNA #1 transfected cells. Arrow points to the 39.2 kD protein and its suppression in siRNA transfected cells. (B) Decreased intensities of protein peak (arrows) in control siRNA transfected cells.
Mentions: The data were further analysed by Biomarker Wizard Software, a univariate analysis tool. After analysis of the intensity values for all spectra at a given mass, there were significant differences in distribution of proteins in the control siRNA transfected ASPC-1 cell culture supernatants compared to the ADAM8 siRNA transfected ASPC-1 supernatants. It could be shown (Fig. 6A) that the peak at m/z 39,232 was more than three-fold higher in control super-natants (height from baseline 2.8) compared to siRNA supernatants (height from baseline 0.8). The lower intensity of this protein in ASPC-1 ADAM8 siRNA transfected cell culture supernatants leads to the conclusion that it is probably a shedded part of ADAM8. The differential profiling of ADAM8 and control transfected cell supernatant spectra revealed in the controls not only a decreased intensity of some protein peaks but also a complete disappearance of some proteins, as shown in the boxed area covering the 5–7 kD range (Fig. 6B). The presence of additional peaks in supernatants of the ADAM8 siRNA trans-fected cell cultures indicates the knock-down ADAM8 expression, and consequently a lower proteolytic activity in these cell culture supernatants.

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