Figure 4

Metastatic markers TGFβ1 and MMP9 are overexpressed in poor outcome HL patients and by HL cell lines. (A) Protein and mRNA expression levels of TGFβ1 and MMP9 in normal lymph node control (NC), good outcome (GO) group and poor outcome (PO) group analyzed by immunostaining (left, images only for PO group) and qRT-PCR (right). mRNA expression is represented by fold-change (y-axis) after normalization with the control (NC). Significance of all qRT-PCR data comparing GO and PO is indicated (p < 0.005). TGFβ1 and MMP9 are also overexpressed by the HL cell lines (lower image of gel electrophoresis of (A)). (B) TGFβ1 and MMP9 protein coexpression in tissues from the poor outcome HL patient group analyzed by double immunofluorescence staining for CD30, TGFβ1 and MMP9, or SDC1, TGFβ1 and MMP9. Individual green or red fluorescence is depicted at the bottom of each image. (C) Coexpression of TGFβ1 and MMP9 by subsets of tumor cells in poor outcome sample. (Inset of A and B) Hodgkin Reed Sternberg cells (HRS) coexpressing SDC1 and TGFβ1 or SDC1 and MMP9. Scale bar (white solid bar) represents 100 μm.