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Type IV collagen is a tumour stroma-derived biomarker for pancreas cancer.

Ohlund D, Lundin C, Ardnor B, Oman M, Naredi P, Sund M - Br. J. Cancer (2009)

Bottom Line: The field is hampered by a lack of good prognostic and predictive tumour biomarkers to be used during follow-up of patients.Persisting high levels of circulating type IV collagen after surgery indicates a quick relapse in disease and poor survival.Our results most importantly show that stroma related substances can be evaluated as potential cancer biomarkers, and thereby underline the importance of the tumour microenvironment also in this context.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical and Perioperative Sciences, Surgery, Umeå University, Umeå, Sweden.

ABSTRACT

Background: Pancreas cancer is a dreaded disease with high mortality, despite progress in surgical and oncological treatments in recent years. The field is hampered by a lack of good prognostic and predictive tumour biomarkers to be used during follow-up of patients.

Methods: The circulating level of type IV collagen was measured by ELISA in pancreas cancer patients and controls. The expression pattern of type IV collagen in normal pancreas, pancreas cancer tissue and in pancreas cancer cell lines was studied by immunofluorescence and Western blot techniques.

Results: Patients with pancreas cancer have significantly increased circulating levels of type IV collagen. In pancreas cancer tissue high levels of type IV collagen expression was found in close proximity to cancer cells in the tumour stroma. Furthermore, pancreas cancer cells were found to produce and secrete type IV collagen in vitro, which in part can explain the high type IV collagen expression observed in pancreas cancer tissue, and the increased circulating levels in pancreas cancer patients. Of clinical importance, our results show that the circulating level of type IV collagen after surgery is strongly related to prognosis in patients treated for pancreas cancer by pancreatico-duodenectomy with curative intent. Persisting high levels of circulating type IV collagen after surgery indicates a quick relapse in disease and poor survival.

Conclusion: Our results most importantly show that stroma related substances can be evaluated as potential cancer biomarkers, and thereby underline the importance of the tumour microenvironment also in this context.

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Related in: MedlinePlus

Expression of type IV collagen in pancreas cancer cell lines. (A) Cell lysates of HPAC and CFPAC-1 contain type IV collagen-derived fragments, demonstrating production of this collagen by cells. The complete α(IV)-chain is too large to enter the gel, and the bands visualised correspond to fragments of different sizes cleaved off from the α-chain in a constantly ongoing turnover process involving different tumour-associated proteases. In the cell media type IV collagen-derived fragments could also be observed in both cell lines, indicating secretion of type IV collagen from the cells to the media. As positive controls recombinant human arresten (α1(IV)NC1) and canstatin (α2(IV)NC1) were used. As negative control HPAC cell lysate and cell media were incubated without primary antibody. (B–D) Immunocytochemical analysis of HPAC and CFPAC-1 cells confirm that type IV collagen is produced by pancreas cancer cell lines. In (B) type IV collagen is visualised using a polyclonal antibody, in (C) an antibody specific for the α1(IV)-chain is used and in (D) an antibody specific for the α2(IV)-chain. Type IV collagen is shown in red and cell nuclei in blue (DAPI). Magnification × 40 in B–D.
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fig3: Expression of type IV collagen in pancreas cancer cell lines. (A) Cell lysates of HPAC and CFPAC-1 contain type IV collagen-derived fragments, demonstrating production of this collagen by cells. The complete α(IV)-chain is too large to enter the gel, and the bands visualised correspond to fragments of different sizes cleaved off from the α-chain in a constantly ongoing turnover process involving different tumour-associated proteases. In the cell media type IV collagen-derived fragments could also be observed in both cell lines, indicating secretion of type IV collagen from the cells to the media. As positive controls recombinant human arresten (α1(IV)NC1) and canstatin (α2(IV)NC1) were used. As negative control HPAC cell lysate and cell media were incubated without primary antibody. (B–D) Immunocytochemical analysis of HPAC and CFPAC-1 cells confirm that type IV collagen is produced by pancreas cancer cell lines. In (B) type IV collagen is visualised using a polyclonal antibody, in (C) an antibody specific for the α1(IV)-chain is used and in (D) an antibody specific for the α2(IV)-chain. Type IV collagen is shown in red and cell nuclei in blue (DAPI). Magnification × 40 in B–D.

Mentions: To further study whether the observed type IV collagen is solely expressed by the stromal cells, such as the stellate cells surrounding the cancer cells in the tumour, or partly by the cancer cells themselves, we performed in vitro studies on two well characterised pancreas cancer cell lines HPAC and CFPAC-1. Western blot analysis of cell lysate and media, and immunocytochemical staining, show that both α1(IV) and α2(IV), the two α-chains that were strongly expressed in the tumour stroma in pancreas cancer tissue, are produced by the cancer cells and secreted to the media in both cell lines (Figure 3). These results suggest that the type IV collagen observed in the tumour stroma in vivo could be, at least in part, produced by the pancreas cells themselves.


Type IV collagen is a tumour stroma-derived biomarker for pancreas cancer.

Ohlund D, Lundin C, Ardnor B, Oman M, Naredi P, Sund M - Br. J. Cancer (2009)

Expression of type IV collagen in pancreas cancer cell lines. (A) Cell lysates of HPAC and CFPAC-1 contain type IV collagen-derived fragments, demonstrating production of this collagen by cells. The complete α(IV)-chain is too large to enter the gel, and the bands visualised correspond to fragments of different sizes cleaved off from the α-chain in a constantly ongoing turnover process involving different tumour-associated proteases. In the cell media type IV collagen-derived fragments could also be observed in both cell lines, indicating secretion of type IV collagen from the cells to the media. As positive controls recombinant human arresten (α1(IV)NC1) and canstatin (α2(IV)NC1) were used. As negative control HPAC cell lysate and cell media were incubated without primary antibody. (B–D) Immunocytochemical analysis of HPAC and CFPAC-1 cells confirm that type IV collagen is produced by pancreas cancer cell lines. In (B) type IV collagen is visualised using a polyclonal antibody, in (C) an antibody specific for the α1(IV)-chain is used and in (D) an antibody specific for the α2(IV)-chain. Type IV collagen is shown in red and cell nuclei in blue (DAPI). Magnification × 40 in B–D.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2713699&req=5

fig3: Expression of type IV collagen in pancreas cancer cell lines. (A) Cell lysates of HPAC and CFPAC-1 contain type IV collagen-derived fragments, demonstrating production of this collagen by cells. The complete α(IV)-chain is too large to enter the gel, and the bands visualised correspond to fragments of different sizes cleaved off from the α-chain in a constantly ongoing turnover process involving different tumour-associated proteases. In the cell media type IV collagen-derived fragments could also be observed in both cell lines, indicating secretion of type IV collagen from the cells to the media. As positive controls recombinant human arresten (α1(IV)NC1) and canstatin (α2(IV)NC1) were used. As negative control HPAC cell lysate and cell media were incubated without primary antibody. (B–D) Immunocytochemical analysis of HPAC and CFPAC-1 cells confirm that type IV collagen is produced by pancreas cancer cell lines. In (B) type IV collagen is visualised using a polyclonal antibody, in (C) an antibody specific for the α1(IV)-chain is used and in (D) an antibody specific for the α2(IV)-chain. Type IV collagen is shown in red and cell nuclei in blue (DAPI). Magnification × 40 in B–D.
Mentions: To further study whether the observed type IV collagen is solely expressed by the stromal cells, such as the stellate cells surrounding the cancer cells in the tumour, or partly by the cancer cells themselves, we performed in vitro studies on two well characterised pancreas cancer cell lines HPAC and CFPAC-1. Western blot analysis of cell lysate and media, and immunocytochemical staining, show that both α1(IV) and α2(IV), the two α-chains that were strongly expressed in the tumour stroma in pancreas cancer tissue, are produced by the cancer cells and secreted to the media in both cell lines (Figure 3). These results suggest that the type IV collagen observed in the tumour stroma in vivo could be, at least in part, produced by the pancreas cells themselves.

Bottom Line: The field is hampered by a lack of good prognostic and predictive tumour biomarkers to be used during follow-up of patients.Persisting high levels of circulating type IV collagen after surgery indicates a quick relapse in disease and poor survival.Our results most importantly show that stroma related substances can be evaluated as potential cancer biomarkers, and thereby underline the importance of the tumour microenvironment also in this context.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgical and Perioperative Sciences, Surgery, Umeå University, Umeå, Sweden.

ABSTRACT

Background: Pancreas cancer is a dreaded disease with high mortality, despite progress in surgical and oncological treatments in recent years. The field is hampered by a lack of good prognostic and predictive tumour biomarkers to be used during follow-up of patients.

Methods: The circulating level of type IV collagen was measured by ELISA in pancreas cancer patients and controls. The expression pattern of type IV collagen in normal pancreas, pancreas cancer tissue and in pancreas cancer cell lines was studied by immunofluorescence and Western blot techniques.

Results: Patients with pancreas cancer have significantly increased circulating levels of type IV collagen. In pancreas cancer tissue high levels of type IV collagen expression was found in close proximity to cancer cells in the tumour stroma. Furthermore, pancreas cancer cells were found to produce and secrete type IV collagen in vitro, which in part can explain the high type IV collagen expression observed in pancreas cancer tissue, and the increased circulating levels in pancreas cancer patients. Of clinical importance, our results show that the circulating level of type IV collagen after surgery is strongly related to prognosis in patients treated for pancreas cancer by pancreatico-duodenectomy with curative intent. Persisting high levels of circulating type IV collagen after surgery indicates a quick relapse in disease and poor survival.

Conclusion: Our results most importantly show that stroma related substances can be evaluated as potential cancer biomarkers, and thereby underline the importance of the tumour microenvironment also in this context.

Show MeSH
Related in: MedlinePlus