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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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Invasion and growth capacity of ADAM8 siRNA transfected ASPC-1 cells. (A) Real-time quantitative PCR and (B) immunoblot analysis were performed to analyse the effect of four different ADAM8 siRNA oligonucleotides (#1–#4) compared with control trans-fected ASPC-1 cells. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) Cell proliferation and invasion assays were performed as described in the ‘Materials and methods’ section. ADAM siRNA oligonu-cleotides #1 were utilized. The values are expressed as % of control transfected cells and shown as mean ± SEM obtained from four independent experiments.
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fig05: Invasion and growth capacity of ADAM8 siRNA transfected ASPC-1 cells. (A) Real-time quantitative PCR and (B) immunoblot analysis were performed to analyse the effect of four different ADAM8 siRNA oligonucleotides (#1–#4) compared with control trans-fected ASPC-1 cells. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) Cell proliferation and invasion assays were performed as described in the ‘Materials and methods’ section. ADAM siRNA oligonu-cleotides #1 were utilized. The values are expressed as % of control transfected cells and shown as mean ± SEM obtained from four independent experiments.

Mentions: Members of the ADAM family, including ADAM8, are associated with cellular invasion. To investigate the influence of ADAM8 on growth and invasion of pancreatic cancer cells, gene silencing was performed with siRNA oligonucleotides, followed by growth and invasion assays. ASPC-1 pancreatic cancer cells were selected for siRNA transfection experiments since they expressed one of the highest levels of ADAM8 mRNA and protein. The maximum effects of siRNA transfection were observed after 96 hrs, resulting in an 83% reduction of ADAM8 mRNA levels and a 74% reduction in protein levels (Fig. 5A, 5B). Interestingly, silencing of ADAM8 did not significantly influence anchorage-dependent cell growth (Fig. 5C). In contrast, there was significantly reduced invasion (to 36% of control, P < 0.05) (Fig. 5C).


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)

Invasion and growth capacity of ADAM8 siRNA transfected ASPC-1 cells. (A) Real-time quantitative PCR and (B) immunoblot analysis were performed to analyse the effect of four different ADAM8 siRNA oligonucleotides (#1–#4) compared with control trans-fected ASPC-1 cells. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) Cell proliferation and invasion assays were performed as described in the ‘Materials and methods’ section. ADAM siRNA oligonu-cleotides #1 were utilized. The values are expressed as % of control transfected cells and shown as mean ± SEM obtained from four independent experiments.
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Related In: Results  -  Collection

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

fig05: Invasion and growth capacity of ADAM8 siRNA transfected ASPC-1 cells. (A) Real-time quantitative PCR and (B) immunoblot analysis were performed to analyse the effect of four different ADAM8 siRNA oligonucleotides (#1–#4) compared with control trans-fected ASPC-1 cells. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) Cell proliferation and invasion assays were performed as described in the ‘Materials and methods’ section. ADAM siRNA oligonu-cleotides #1 were utilized. The values are expressed as % of control transfected cells and shown as mean ± SEM obtained from four independent experiments.
Mentions: Members of the ADAM family, including ADAM8, are associated with cellular invasion. To investigate the influence of ADAM8 on growth and invasion of pancreatic cancer cells, gene silencing was performed with siRNA oligonucleotides, followed by growth and invasion assays. ASPC-1 pancreatic cancer cells were selected for siRNA transfection experiments since they expressed one of the highest levels of ADAM8 mRNA and protein. The maximum effects of siRNA transfection were observed after 96 hrs, resulting in an 83% reduction of ADAM8 mRNA levels and a 74% reduction in protein levels (Fig. 5A, 5B). Interestingly, silencing of ADAM8 did not significantly influence anchorage-dependent cell growth (Fig. 5C). In contrast, there was significantly reduced invasion (to 36% of control, P < 0.05) (Fig. 5C).

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