Transforming Growth Factor-β3 Increases the Invasiveness of Endometrial Carcinoma Cells through Phosphatidylinositol 3-Kinase-dependent Up-regulation of X-linked Inhibitor of Apoptosis and Protein Kinase C-dependent Induction of Matrix Metalloproteinase-9

Abstract

Tumor cells often acquire intrinsic resistance to the growth inhibitory and pro-apoptotic effects of transforming growth factor-␤ (TGF-␤); moreover, TGF-␤ can confer invasive properties to established tumor cells. In the present study, we show that TGF-␤ isoforms (TGF-␤1, TGF-␤2, and TGF-␤3) trigger proper Smad signaling in human endometrial carcinoma cell lines and efficiently inhibit cellular proliferation. These cells, however, exhibit a high degree of resistance to TGF-␤ pro-apoptotic effects; we found that this resistant phenotype would be acquired through up-regulation of X-linked inhibitor of apoptosis protein (XIAP) levels. In addition, using RNA interference and pharmacological inhibitors, we show that TGF-␤ increases cellular invasiveness via two distinct signaling pathways in endometrial carcinoma cells: phosphatidylinositol 3-kinase/ AKT-dependent up-regulation of XIAP and protein kinase Cdependent induction of matrix-metalloproteinase-9 (MMP-9) expression. Additionally, these findings were correlated with clinical observations showing abundant TGF-␤ immunoreactivity in human endometrial carcinoma tumors in vivo, extending from the epithelial compartment to the stroma upon acquisition of an invasive phenotype (gradually from grades I to III). Collectively our results describe for the first time a role for TGF-␤3 in tumor invasiveness.

Progressive pulmonary fibrosis is mediated by TGF-β isoform 1 but not TGF-β3

“…This is in line with a previous report indicating that TGF-β1 is a more potent inductor of TIMP-1 compared to TGF-β3 in human lung fibroblasts, which is consistent with the findings of our study [see figure 8 in (Eickelberg et al, 1999)]. Further, exogenous TGF-β3, but not TGF-β1 increased MMP-9 expression and activity in cancer cells and a lip wound healing model (Hosokawa et al, 2003;Van Themsche et al, 2007), indicating a role in balancing protease/ antiprotease levels. However, there is also a study suggesting that TGF-β3 may be an even more potent inducer of procollagen and inhibitor of matrix degrading activity than TGF-β1 and -β2 (Coker et al, 1997).…”

“…Others have shown a differential expression pattern of protease and antiprotease levels. TGF-β3 alone was not sufficient to induce TIMP-1 expression in intestinal myofibroblasts (McKaig, McWilliams, Watson, & Mahida, 2003) or human endometrial carcinoma cells (Van Themsche et al, 2007). This is in line with a previous report indicating that TGF-β1 is a more potent inductor of TIMP-1 compared to TGF-β3 in human lung fibroblasts, which is consistent with the findings of our study [see figure 8 in (Eickelberg et al, 1999)].…”

“…Some of these studies supported no major individual role, and claimed that TGF-β3 acts in concert with TGF-β1 (Eickelberg et al, 1999;Khalil, Shing, & Whitman, 1993;Santana, Saxena, Noble, Gold, & Marshall, 1995). In contrast, data from targeted gene knockouts and experimental models of cutaneous wound healing and chronic inflammatory bowel disease suggested distinct features of TGF-β3, when compared with TGF-β1 (Ingman & Robertson, 2002;McKaig, Hughes, Tighe, & Mahida, 2002;Shah, Foreman, & Ferguson, 1995;Van Themsche, Mathieu, Parent, & Asselin, 2007). Fetal wounds, which contain primarily TGF-β3, heal without scars, whereas adult wounds, which contain mainly TGF-β1 and -β2, always exhibit some degree of scarring (Nath, LaRegina, Markham, Ksander, & Weeks, 1994).…”

Keratin 8 and 18 Loss in Epithelial Cancer Cells Increases Collective Cell Migration and Cisplatin Sensitivity through Claudin1 Up-regulation

“…Immunoblotting and Immunoprecipitation-Cell lysis, electrophoresis, and Western blotting were performed as described previously (31). All of the antibodies were from Cell Signaling Technology except for K8 (Troma1, Developmental Studies Hybridoma Bank, University of Iowa); K18 (L2A1) (32), which was a generous gift from Dr. M. Bishr Omary (University of Michigan); Akt3 (Millipore, Billerica, MA); horseradish peroxidase (HRP)-conjugated ␤-actin (Sigma); ␤-tubulin (Abcam Inc., Cambridge, MA); and HRP-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA).…”

“…Conventional RT-PCR-Total RNA extraction, cDNA synthesis, and PCRs were performed as described previously (31). The conditions for PCRs are listed in Table 1.…”