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. 2023 Mar 13;13:1107021. doi: 10.3389/fonc.2023.1107021

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Copyright © 2023 Wiesehöfer, Raczinski, Wiesehöfer, Dankert, Czyrnik, Spahn, Kruithof-de Julio and Wennemuth

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Proteases, shedding the ectodomain of EREG are increased in CRPC compared to CSPC cells. (A) The dot plot depicts the ADAM17 expression of CSPC and CRPC cell lines using qRT-PCR analysis. 22Rv1, DU145, PC3 and LNCaP_EnzR (p<0.0001) significantly express more ADAM17 compared to LNCaP cells. (B) Flow cytometry analysis using ADAM17 antibody show the ADAM17 presence on the cell surface of CSPC and CRPC cell lines. ADAM17 protein level on cell surface of PC3 (p<0.0001) and LNCaP_EnzR (p < 0.0001) cells is increased in comparison to LNCaP cells whereas ADAM17 protein content on 22Rv1 and DU145 is not altered. (C) The pictures show representative flow cytometry analysis of CSPC and CRPC cell lines using ADAM17 antibody (dark) and mouse-IgG (light) as a control. MMP2 (D) and MMP9 (E) expression in CSPC and CRPC cell lines was determined by qRT-PCR. In comparison to LNCaP cells 22Rv1 (p=0.0057), PC3 (p=0.0002) and LNCaP_EnzR (p < 0.0001) cells show an increased expression of MMP2. The MMP9 expression of 22Rv1 (p < 0.0001), DU145 (p < 0.0001), PC3 (p < 0.0001) and LNCaP_EnzR (p < 0.0001) is enhanced compared to LNCaP cells. (F) Solid phase ELISA using a specific MMP2 antibody demonstrates the secretion of MMP2 in cell culture supernatant of PCa cell lines. The ELISA results reveal that MMP2 is secreted by all five PCa cell lines, but 22Rv1 (p=0.0021), DU145 (p=0.0026), PC3 (p < 0.0001) and LNCaP_EnzR (p < 0.0001) cells secrete a higher amount of MMP2 protein in comparison to LNCaP cells. (G) PCa cell culture supernatant was collected and MMP9 protein was investigated by solid phase ELISA. All PCa cells secrete MMP9 protein, but the CRPC cell lines 22Rv1 (p < 0.0001), DU145 (p=0.0005), PC3 (p < 0.0001) and LNCaP_EnzR (p < 0.0001) secrete more MMP9 protein than CSPC cell line LNCaP. All graphs show the mean and SD from four independent experiments (ns, not significant; **p < 0.01; ***p < 0.001; ****p < 0.0001).