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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 under normoxic and hypoxic conditions. ADAM8 mRNA (A) and protein (B) expression in pancreatic cancer cell lines under normal and hypoxic conditions (24 hrs) was determined by qRT-PCR and immunoblotting, respectively. The response towards hypoxic conditions was monitored by HIF-1α expression levels. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) ELISA: ADAM8 expression in the cell lysates (grey bars) and cell culture supernatants (white bars). The values are presented as mean +/− SEM.
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fig04: ADAM8 expression under normoxic and hypoxic conditions. ADAM8 mRNA (A) and protein (B) expression in pancreatic cancer cell lines under normal and hypoxic conditions (24 hrs) was determined by qRT-PCR and immunoblotting, respectively. The response towards hypoxic conditions was monitored by HIF-1α expression levels. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) ELISA: ADAM8 expression in the cell lysates (grey bars) and cell culture supernatants (white bars). The values are presented as mean +/− SEM.

Mentions: Next we sought to examine the regulation of ADAM8 mRNA and protein expression in cultured pancreatic cancer cell lines under the effects of hypoxia as has been shown previously for other ADAMs [22, 23]. In addition we sought to test the effects of ADAM8 silencing on pancreatic cancer cell growth, invasion and potential target proteins. Therefore, mRNA levels of ADAM8 were determined in these cells. Different amounts of ADAM8 mRNA with a range of 16–922 copies/μl cDNA were detected in all examined pancreatic cancer cell lines (Fig. 4A). Immunoblot analysis was performed to compare the mRNA data with the corresponding protein expression. As in pancreatic cancer tissues, ADAM8 was present in two forms (60 kD and 85–95 kD) in the investigated cell lines (Fig. 4B). These results were confirmed by ELISA, which demonstrated ADAM8 expression in all pancreatic cancer cell lysates (Fig. 4C). In contrast, the released ADAM8 ectodomain was detectable in only four of eight pancreatic cancer cell supernatants (Fig. 4C).


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 under normoxic and hypoxic conditions. ADAM8 mRNA (A) and protein (B) expression in pancreatic cancer cell lines under normal and hypoxic conditions (24 hrs) was determined by qRT-PCR and immunoblotting, respectively. The response towards hypoxic conditions was monitored by HIF-1α expression levels. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) ELISA: ADAM8 expression in the cell lysates (grey bars) and cell culture supernatants (white bars). The values are presented as mean +/− SEM.
© Copyright Policy
Related In: Results  -  Collection

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

fig04: ADAM8 expression under normoxic and hypoxic conditions. ADAM8 mRNA (A) and protein (B) expression in pancreatic cancer cell lines under normal and hypoxic conditions (24 hrs) was determined by qRT-PCR and immunoblotting, respectively. The response towards hypoxic conditions was monitored by HIF-1α expression levels. Equal loading of the protein samples was confirmed using an -tubulin antibody. Size markers are indicated on the left (in kDa). (C) ELISA: ADAM8 expression in the cell lysates (grey bars) and cell culture supernatants (white bars). The values are presented as mean +/− SEM.
Mentions: Next we sought to examine the regulation of ADAM8 mRNA and protein expression in cultured pancreatic cancer cell lines under the effects of hypoxia as has been shown previously for other ADAMs [22, 23]. In addition we sought to test the effects of ADAM8 silencing on pancreatic cancer cell growth, invasion and potential target proteins. Therefore, mRNA levels of ADAM8 were determined in these cells. Different amounts of ADAM8 mRNA with a range of 16–922 copies/μl cDNA were detected in all examined pancreatic cancer cell lines (Fig. 4A). Immunoblot analysis was performed to compare the mRNA data with the corresponding protein expression. As in pancreatic cancer tissues, ADAM8 was present in two forms (60 kD and 85–95 kD) in the investigated cell lines (Fig. 4B). These results were confirmed by ELISA, which demonstrated ADAM8 expression in all pancreatic cancer cell lysates (Fig. 4C). In contrast, the released ADAM8 ectodomain was detectable in only four of eight pancreatic cancer cell supernatants (Fig. 4C).

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