Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression

Webber, Mukta M.; Quader, Salmaan T.A.; Kleinman, Hynda K.; Deocampo, Daniel M.; Storto, Patrick D.; Bice, Gillian; DeMendonca‐Calaca, Wanderley; Williams, Daniel E.

doi:10.1002/pros.1041.abs

Cited by 25 publications

(44 citation statements)

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“…BPH-1 can be induced to form tumors by various factors, such as association with a tumorigenic stromal microenvironment, treatment with hormonal carcinogens or by oncogene transformation (31,32). In this study, we found that the increase in O-GlcNAcylation by thiamet-G induced anchorage-independent growth and migration in BPH-1 cells, which is the first direct evidence that O-GlcNAc can induce malignant transformation in bening cells.…”

Section: Discussionsupporting

confidence: 51%

O-GlcNAcylation is increased in prostate cancer tissues and enhances malignancy of prostate cancer cells

Gao

Han

et al. 2014

Molecular Medicine Reports

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Abstract. O-GlcNAc is an O-linked β-N-acetylglucosamine moiety attached to the side-chain hydroxyl of a serine or threonine residue in numerous cytoplasmic and nuclear proteins. In this study, we detected the level of O-GlcNAc in prostate, liver and pancreatic cancer tissues, and found that the global O-GlcNAc modification also known as O-GlcNAcylation, is specifically increased in prostate cancer tissues compared to corresponding adjacent tissues. In addition, we found that global O-GlcNAcylation is increased in prostate cancer cells and not in benign prostatic hyperplasia (BPH) epithelial cells. O-GlcNAc enhanced the anchorage-independent growth and the migratory/invasive ability of prostate cancer cells. More importantly, we provide here, for the first time to the best of our knowledge, direct evidence that increased O-GlcNAcylation induces malignant transformation of nontumorigenic (BPH) cells. Furthermore, our study suggested that inhibiting the formation of the E-cadherin/catenin/cytoskeleton complex may underly the O-GlcNAc-induced prostate cancer progression. Overall, these findings indicated that O-GlcNAcylation is increased in prostate, but not in liver and pancreatic cancer tissues, and that O-GlcNAc can enhance the malignancy of prostate cancer cells. IntroductionNumerous nuclear and cytoplasmic proteins have been found modified with O-β-N-acetylglucosamine (O-GlcNAc) at the hydroxyl moiety of serine or threonine residues. This moiety is dynamically added and removed by the O-GlcNAc transferase (OGT) and the O-GlcNAcase (OGA) enzymes, respectively (1). UDP-GlcNAc is the donor substrate of OGT, and is biosynthesized through the hexosamine pathway (HBP). The HBP flux is highly dependent on glucose and glutamine, with ~3-5% of total glucose entering this pathway (2). O-GlcNAc is a sensor of intracellular glucose metabolism, since the intracellular level of UDP-GlcNAc is the main regulatory factor for the activity of OGT (3). O-GlcNAcylation is altered in metabolism-associated diseases, such as type II diabetes (4-6), Alzheimer's disease (7) and cancer (8). Therefore, abnormal O-GlcNAcylation may play critical roles in these pathological processes.Aberrant metabolism is a hallmark of cancer. Most cancer cells show increased rates of glucose and glutamine utilization, up to 200-fold higher than those observed in the healthy cells they originate from, and predominantly produce energy through glycolysis followed by lactic acid fermentation (9,10). As a glycolytic pathway, HBP can enhance the level of UDP-GlcNAc, and thus, the activity of OGT, which may lead to an increase in the global O-GlcNAc level (summed over all proteins) in cancer cells. We have previously demonstrated that the global O-GlcNAc level is increased in breast, lung and colon cancer tissues as compared to respective adjacent tissues (11,12). In addition, other research groups have shown that O-GlcNAcylation is increased in breast, prostate and pancreatic cancer cell lines (13-15). However, whether increased O-GlcNAcylation is universal in ...

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Section: Discussionsupporting

confidence: 51%

O-GlcNAcylation is increased in prostate cancer tissues and enhances malignancy of prostate cancer cells

Gao

Han

et al. 2014

Molecular Medicine Reports

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“…In order to examine the role of Dkk-3 in more detail, we established a cell culture model using RWPE-1, a non-malignant immortalized human prostatic epithelial cell line that is used as a physiologically relevant system for the regulation of growth, morphogenesis and differentiation in the normal human prostate (Kawano et al, 2006;Webber et al, 1997;Webber et al, 2001). RWPE-1 cells are immortalized by human papillomavirus 18, retain expression of luminal epithelial cell markers, such as cytokeratin 8 and 18 , and have the ability to form acini with a hollow lumen when grown in 3D Matrigel cultures (Bello-DeOcampo et al, 2001a), a feature that is not manifested by LNCaP (Härmä et al, 2010) or DU145 (BelloDeOcampo et al, 2001b) Fig.…”

Section: Resultsmentioning

confidence: 99%

Downregulation of Dickkopf-3 disrupts prostate acinar morphogenesis through TGF-β/Smad signaling

Romero

Kawano

Bengoa³

et al. 2013

Journal of Cell Science

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SummaryLoss of tissue organization is a hallmark of the early stages of cancer, and there is considerable interest in proteins that maintain normal tissue architecture. Prostate epithelial cells cultured in Matrigel form three-dimensional acini that mimic aspects of prostate gland development. The organization of these structures requires the tumor suppressor Dickkopf-3 (Dkk-3), a divergent member of the Dkk family of secreted Wnt signalling antagonists that is frequently downregulated in prostate cancer. To gain further insight into the function of Dkk-3 in the prostate, we compared the prostates of Dkk3-null mice with those of control littermates. We found increased proliferation of prostate epithelial cells in the mutant mice and changes in prostate tissue organization. Consistent with these observations, cell proliferation was elevated in acini formed by human prostate epithelial cells stably silenced for Dkk-3. Silencing of Dkk-3 increased TGF-b/Smad signalling, and inhibitors of TGF-b/Smad signalling rescued the defective acinar phenotype caused by loss of Dkk-3. These findings suggest that Dkk-3 maintains the structural integrity of the prostate gland by limiting TGF-b/Smad signalling.

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“…Both are malignant, tumorigenic cell lines of human prostate epithelial origin. NB11 cells were cultured as described previously (30), in complete keratinocyte serum-free medium (Invitrogen) containing 50 g/ml bovine pituitary extract (Invitrogen), 5 ng/ml epidermal growth factor supplement (Invitrogen), and 100 g/ml gentamicin (Invitrogen). LNCaP cells were maintained in RPMI 1640 (Invitrogen) containing 10% fetal bovine serum (Invitrogen) and 100 g/ml gentamicin until cells reached confluence, at which time the medium was replaced with serum-free RPMI 1640.…”

Section: Methodsmentioning

confidence: 99%

The Amyloid Precursor Protein/Protease Nexin 2 Kunitz Inhibitor Domain Is a Highly Specific Substrate of Mesotrypsin

Salameh

Robinson

Navaneetham

et al. 2010

Journal of Biological Chemistry

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The amyloid precursor protein (APP) is a ubiquitously expressed transmembrane adhesion protein and the progenitor of amyloid-␤ peptides. The major splice isoforms of APP expressed by most tissues contain a Kunitz protease inhibitor domain; secreted APP containing this domain is also known as protease nexin 2 and potently inhibits serine proteases, including trypsin and coagulation factors. The atypical human trypsin isoform mesotrypsin is resistant to inhibition by most protein protease inhibitors and cleaves some inhibitors at a substantially accelerated rate. Here, in a proteomic screen to identify potential physiological substrates of mesotrypsin, we find that APP/protease nexin 2 is selectively cleaved by mesotrypsin within the Kunitz protease inhibitor domain. In studies employing the recombinant Kunitz domain of APP (APPI), we show that mesotrypsin cleaves selectively at the Arg 15 -Ala 16 reactive site bond, with kinetic constants approaching those of other proteases toward highly specific protein substrates. Finally, we show that cleavage of APPI compromises its inhibition of other serine proteases, including cationic trypsin and factor XIa, by 2 orders of magnitude. Because APP/protease nexin 2 and mesotrypsin are coexpressed in a number of tissues, we suggest that processing by mesotrypsin may ablate the protease inhibitory function of APP/protease nexin 2 in vivo and may also modulate other activities of APP/protease nexin 2 that involve the Kunitz domain.Mesotrypsin is a human trypsin encoded by the PRSS3 gene found on chromosome 9p13 (1). Normal expression of PRSS3 is restricted to pancreas, brain, and, to a lesser extent, small intestine and colon (2-4); additionally, PRSS3 appears to be transcriptionally up-regulated with cancer progression in epithelial cancers, including lung (5), colon (6), and prostate. Mesotrypsin exhibits substantially different specificity from other trypsins toward protein substrates. It fails to activate pancreatic zymogens and also shows reduced capacity to degrade trypsinogens (7). Compared with other trypsins, it is significantly compromised in its ability to cleave protease-activated receptors (8 -10). Despite limited activity toward these classic trypsin substrates, mesotrypsin displays enhanced catalytic activity compared with other trypsins in the cleavage of certain specific protein substrates, most notably several canonical protease inhibitors (7, 11).The "canonical" inhibitors of serine proteases, named for a protease-binding loop of highly characteristic backbone conformation (12, 13), fulfill the paradoxical function of binding to a protease in a substrate-like manner yet acting as an inhibitor rather than an ordinary substrate. These inhibitors, representing at least 18 different convergently evolved protein families (14, 15), inhibit their cognate proteases via the "Laskowski mechanism," in which inhibitors act as highly specific, limited proteolysis substrates for target enzymes (14, 16). They bind so as to position a specific peptide bond, the "reactive si...

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Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression

Cited by 25 publications

References 0 publications

O-GlcNAcylation is increased in prostate cancer tissues and enhances malignancy of prostate cancer cells

O-GlcNAcylation is increased in prostate cancer tissues and enhances malignancy of prostate cancer cells

Downregulation of Dickkopf-3 disrupts prostate acinar morphogenesis through TGF-β/Smad signaling

The Amyloid Precursor Protein/Protease Nexin 2 Kunitz Inhibitor Domain Is a Highly Specific Substrate of Mesotrypsin

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