Primary versus castration-resistant prostate cancer: modeling through novel murine prostate cancer cell lines

Cell lines representing the progression of prostate cancer (PC) from an androgen-dependent to an androgen-independent state are scarce. In this study, we used previously characterized prostate luminal epithelial cell line (Plum), under androgen influence, to establish cellular models of PC progression. Cells derived from orthotopic tumors have been isolated to develop an androgen-dependent (PLum-AD) versus an androgen-independent (PLum-AI) model. Upon immunofluorescent, qRT-PCR and Western blot analyses, PLum-AD cells mostly expressed prostate epithelial markers while PLum-AI cells expressed mesenchymal cell markers. Interestingly, both cell lines maintained a population of stem/progenitor cells. Furthermore, our data suggest that both cell lines are tumorigenic; PLum-AD resulted in an adenocarcinoma whereas PLum-AI resulted in a sarcomatoid carcinoma when transplanted subcutaneously in NOD-SCID mice. Finally, gene expression profiles showed enrichment in functions involved in cell migration, apoptosis, as well as neoplasm invasiveness and metastasis in PLum-AI cells. In conclusion, these data suggest that the newly isolated cell lines represent a new in vitro model of androgen-dependent and –independent PC.

In addition, site-directed protease-null mutant high temperature requirement A2S306A expression blocked apoptosis induced by ITGA7., In this study, expression of the a7 integrin transgene did suppress apoptosis in a7BX2-mdx/ utr -/-mice compared to mdx/utr -/-mice., Concomitantly, integrin a7 silencing promoted apoptotic signaling and made airway smooth muscle cells refractory to the suppressive effects of serum deprivation on caspase 3 activity (Fig. 3C PDGFRA --+> cell migration

Regulation
The PDGF alpha-receptor also mediates a mitogenic signal, but fails to induce cell migration in certain cell types., Activation of the platelet-derived growth factor alpha receptor (alphaPDGFR) leads to cell migration and DNA synthesis., According to our data, mole ovotestes result from a process of PDGFRalpha-mediated mesonephric cell migration, which occurs simultaneously in both sexes., PDGFRA, a receptor whose activity is required for cell migration through the primitive streak, is a target of miR-130b and -218 in vivo. EGLN3 --+> apoptosis Regulation EGLN3 also has pro-apoptotic activity in some cell types., PHD3 may be an important regulator of apoptosis and it is mainly found in tumors with good prognosis., EglN2 and EglN3 also appear to play Hypoxia-inducible factorindependent roles in regulating cell proliferation and apoptosis, respectively., One isoform, PHD3, is expressed in response to hypoxia and causes apoptosis in oxygenated conditions in neural cells. THBS1 --+> cell invasion Regulation TSP-1 up-regulated KB tumor cell invasion 5-fold., Stromal TSP-1 may play a role in regulating tumor cell invasion., Tumor cell invasion was upregulated 3.5 to 4.5-fold by TSP-1 and TGF-beta 1, respectively., Antibodies against uPA or uPAR neutralized the TSP-1-and TGF-beta 1-promoted breast tumor cell invasion., TSP-1 promotes tumor cell invasion of collagen by breast cancer cells., HGF stimulation of cell invasion and enhancement of MMP-9 expression were partially suppressed by TSP-1 overexpression., We previously showed that thrombospondin 1 (TSP-1) upregulates the plasminogen/plasmin system and promotes breast tumor cell ST14 --+> cell migration

Regulation
The serine protease MT-SP1/matriptase plays an important role in cell migration and matrix degradation., Furthermore, TGF-beta-induced cell migration and invasion were significantly impaired by epithin knockdown., These data suggest that matriptase might be a central regulator of cell migration and cancer invasion., These studies demonstrated that matriptase, likely its protease activity, is critical for neural progenitor cell migration., These results further support our working hypothesis that matriptase is an upstream regulator of cellular migration and extracellular matrix degradation. <more data available. CLDN7 ---| cell migration Regulation Taken together, our study demonstrates that claudin-7 inhibits cell migration and invasion through ERK/MAPK signaling pathway in response to growth factor stimulation in human lung cancer cells., Furthermore, claudin-6, 7, and 9 enhance gastric adenocarcinoma cell migration and invasion., Our microarray analysis provides several possible mechanisms by which claudin-7 can affect cell migration and invasion, and these different mechanisms are currently being investigated., Claudin-7 overexpression inhibits cell migration and invasion in NCI-H1299, a human non-small cell lung cancer cell line that lacks endogenous claudin-7 expression. SOD3 ---| apoptosis Regulation EC-SOD is critical for normal cardiac morphology and protects the heart from oxidant-induced fibrosis, apoptosis, and loss of function., In addition, we showed that the thin epidermis of EC-SOD transgenic mice results from the apoptosis of epidermal cells., Extracellular superoxide dismutase (SOD3), which dismutates superoxide anion to hydrogen peroxide, has been shown to reduce the free radical stress derived apoptosis in tissue injuries., Clinical chemistry and tissue pathology analyses showed that adenoviral EC-SOD gene transfer significantly attenuated release of liver enzymes and inhibited necrosis and apoptosis caused by paracetamol overdose. <more data available. HIF1A --+> FLT1 PromoterBinding Conversely, HIF-1alpha knockdown using antisense oligonucleotides, decreased sFlt-1 expression., In contrast, HIF-1a siRNA inhibited the LPS-induced increase in sFlt-1 production (Fig. 4C)., The HIF (hypoxia-inducible factor)-1a transcription factor regulates VEGF and Flt-1 gene transcription [25]., The mRNA levels of VEGF, VEGFR1, Ang-2, and Ang-4 in HPAECs were also increased by either hypoxia or infection with Ad2/HIF-1alpha/VP16 (Figure 3) HIF1A --+> FLT1 MolTransport Furthermore, knockdown of hypoxia-inducible factor-1a inhibited the hypoxia-induced secretion of sFlt-1 and soluble endoglin., Additionally, siRNA-mediated knockdown of HIF-1a inhibited the hypoxia-induced secretion of sFlt-1 and soluble Endoglin., These results indicate that hypoxiainducible factor-1a is a key mediator of the hypoxia-induced secretion of sFlt-1 and soluble endoglin. TGFBR3 ---| cell migration

Regulation
In particular, betaglycan may be essential for inhibinmediated regulation of granulosa cell migration and adhesion, which are integrally involved in the ..., ... betaglycan tonically suppresses nuclear factor-?B-mediated breast cancer cell migration via its intracellular interactions with the scaffolding protein ß-..., Moreover, overexpression of betaglycan resulted in a significant reduction in cell migration whereas inhibin-· gene silencing enhanced both migration and invasion (63)., For instance, Nov, Rgs4 and Tgfbr3, which are inhibitors of cancer cell migration and invasion,48-50 were expressed in increased levels in DA.ACI(Cia10 WNT4 ---| apoptosis Regulation WNT4 may also act to prevent oocyte apoptosis in the developing ovary [35]., Restoring Wnt4 or Wnt5a expression by gene transfection or recombinant protein reduced high glucose-induced cell apoptosis., Wnt4 was previously identified to control the nuclear localization of FOXO1, a transcription factor and known promoter of apoptosis (reviewed in)., Since cell survival during decidualization appears to be regulated by WNT4, we wanted to determine the mechanism by which WNT4 regulates apoptosis., Together with a significant reduction in Spermatogonial stem cell numbers in 5-week-old mutant mouse testes (Fig. 1)

Regulation
In this report, we identified K18 phospho-Ser33 as the site that regulates keratin/14-3-3 binding during mitosis, and showed that K18 Ser33 phosphorylation generates a unique motif that is essential but not sufficient for binding of 14-3-3 proteins.
info Binding Neutrophil gelatinase-associated lipocalcin (NGAL) is an early indicator of kidney injury., Levels of Cystatin-C, as well as serum and urine neutrophil gelatinase-associated lipocain (NGAL), were significantly augmented., This review discusses a member of the lipocalin superfamily, 24p3 or neutrophil gelatinase-associated lipocalcin (NGAL), which induces the formation of kidney epithelia., In this study, levels of interleukin -8, IL-6, interleukin-1 receptor antagonist (IL-1ra), secretory leukocyte protease inhibitor (SLPI), and neutrophil gelatinase-associated lipocalcin (NGAL) were measured in divers before and after a 2-mo period of daily diving. <more data available. Moreover, TSP-1 null hearts had increased myocardial matrix metalloproteinase 3 expression and enhanced matrix metalloproteinase 9 activation after pressure overload., Furthermore, matrix metalloproteinase 3 and 9 are upregulated in Tsp1-knockout mice., Thus, processing and consequent activation of pro-MMP-9 to MMP-9 by physiological activators such as MMP-3 is blocked by TSP-1., thrombospondin-1 may inhibit MMP-9 activity by preventing activation of pro-MMP-2, and MMP-3 can cleave and activate MMP-2 and MMP-9.  (Fig 1G), followed by fixing and processing the same cell for double immunofluorescence (Fig 1H and Fig I) Using protein immunoblotting, we confirmed that MCF7-Six1 lines contained increased levels of the mesenchymal protein fibronectin and decreased levels of the epithelial proteins cytokeratin 18, claudin-1, and claudin-8 ( Figure 3B and Supplemental Figure 2), consistent with a Six1-induced epithelial-mesenchymal transition. Regulation VEGF has been shown to regulate MMP-2 expression in several cell lines.5 Taken together, these findings suggest that swimming training likely provides these beneficial effects on ischemic revascularization in aged mice through VEGF/ Flt-1-mediated MMP-2 activation induced by stimulation of insulin-like growth factor-1/phosphatidylinositol 3-kinase/ Akt-dependent hypoxia-induced factor-1a synthesis and prevention of hypoxia-induced factor-1a destabilization. We previously showed that there was a relation between WT1 and Pdgfra, which may affect the process of epithelial-to-mesenchymal transformation [7]., In addition, platelet-derived growth factor family receptora was shown to be important for maintenance of epithelial-mesenchymal transition, evasion of apoptosis, and metastasis in transformed mesenchymal murine mammary cells. Therefore, nonmetastatic 67NR cells that express a higher mH2A1.1-to-mH2A1.2 ratio than metastatic 4T1 cells might be able to adapt their genetic programs to redox metabolism by expressing genes such as SOD3, which would in turn affect cell invasiveness. LCN2 ----THBS1 Binding DNMT1 is nuclear, but reported to co-localize with annexin V. Lipocalin 2 is decreased by 1.29 fold and is also negatively associated with THBS1, which is increased by 2.13 fold and identified as a binding partner of ELA2. PDGFRA ---> MMP3

Expression
Our studies show that PDGFR-alpha induced MMP3 gene expression and increased cell proliferation and cell migration upon stimulation by platelet-derived growth factor -AA., We investigated PDGF-alpha receptor -and beta receptor -mediated signaling pathways for the expression of MMP-3 and invasion activity using porcine aortic endothelial cells with stable expression of normal or mutated Platelet-derived growth factor receptors.