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. 2009 Jun 2;101(1):91–97. doi: 10.1038/sj.bjc.6605107

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Copyright © 2009 Cancer Research UK

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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.