Expression of two WFDC1/ps20 isoforms in prostate stromal cells induces paracrine apoptosis through regulation of PTGS2/COX-2

Hickman, Oliver; Smith, Richard A.; Rao, Sudha; Nayak, Suraja Kumar; Sreenivasan, Shubha; Vyakarnam, Annapurna; Galustian, Christine

doi:10.1038/bjc.2016.91

Cited by 7 publications

(11 citation statements)

References 31 publications

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“…Increased cytochrome c oxidase subunit II (COX-2) expression has been linked to the initiation and progression of human (11), and COX-2 has been reported to play a key role in the apoptosis of cancer cells (12). In the present study, it was found that miR-26a was significantly downregulated in prostate cancer tissues.…”

Section: Introductionmentioning

confidence: 47%

Downregulated microRNA-26a modulates prostate cancer cell proliferation and apoptosis by targeting COX-2

2016

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Abstract. ) is expressed at lower levels in prostate cancer cells compared with normal prostate cells. However, the regulatory mechanism of miR-26a in tumorigenesis and metastasis is not clear. In the present study, the expression profile of cellular miR-26a was analyzed by reverse transcription-quantitative polymerase chain reaction. The potential target of miR-26a was identified by luciferase assay and western blotting. Examination of miR-26a function was performed by transfection with miR-26a mimics and inhibitor. It was found that miR-26a expression was decreased in prostate cancer tissues and cell lines, with androgen-independent prostate cancer (AIPC) showing lower miR-26a expression compared with androgen-dependent prostate cancer (ADPC). Overexpression of miR-26a by transfecting miR-26a mimics could significantly enhance apoptosis, and this upregulation of apoptosis was triggered by cytochrome c oxidase subunit II inhibition. Furthermore, it was found that lower miR-26a density resulted in an evidently poor prognosis. Understanding the important roles of miR-26a in regulating cell apoptosis in human prostate cancer cells may aid the exploration of AIPC transformation mechanisms and contribute to the development of miRNA-based therapy in the future.

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Section: Introductionmentioning

confidence: 47%

Downregulated microRNA-26a modulates prostate cancer cell proliferation and apoptosis by targeting COX-2

2016

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“…2 (E)). Notably, WB of ps20 preparations including rps20 293F and ps20 HeLa with anti-ps20 Ab to both C-and N-terminal regions revealed the presence of the protein product of an exon 3 truncated WFDC1 mRNA described [6] and previously cloned and expressed by our lab [13] .…”

Section: Discussionmentioning

confidence: 71%

“…2 (B) and (C) arrows, and Supplementary Fig. 1 ) [13] . The third minor band at ≈15 kDa, is only detected by the C-terminal antibody in both rps20 293F and ps20 HeLa preparations ( Fig.…”

Section: Resultsmentioning

confidence: 98%

“…In addition, we have failed to observe any function in V5-His tagged ps20, suggesting that C-terminal tags may be broadly inhibitory to ps20 function. Furthermore, crude CM from cells transfected to express rps20 has repeatedly failed to inhibit cellular proliferation, despite achieving high concentrations [13] . We utilised immunoaffinity chromatography with anti-ps20 1G7 to purify recombinant ps20 from 293F cells (rps20 293F , Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Both were originally observed in prostate cancer cell lines and patient samples [6] . We have recently published evidence that both forms are transcribed and expressed at the protein level [13] . Despite the importance of WFDC family proteins in numerous disease processes, their biochemistry is poorly understood, including their means of interacting with cells and signalling.…”

Section: Introductionmentioning

confidence: 99%

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Cathepsin-L and transglutaminase dependent processing of ps20: A novel mechanism for ps20 regulation via ECM cross-linking

Hickman

Galustian

Smith

et al. 2016

Biochemistry and Biophysics Reports

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Whey-acidic-protein (WAP) four-disulphide core (WFDC) proteins have important roles in the regulation of innate immunity, anti-microbial function, and the inhibition of inflammatory proteases at mucosal surfaces. It was recently demonstrated that the WFDC protein, prostate stromal 20 (ps20), encoded by the WFDC1 gene, is a potent growth inhibitory factor, and shares with other WFDC proteins the ability to modulate wound healing processes and immune responses to viral infections. However, ps20 remains relatively uncharacterised at the protein level.Using a panel of ps20 antibodies for western-blotting (WB), ELISA and immunoaffinity purification, we isolated, biochemically characterised and tested ps20 preparations for three biological properties: (i) interactions with glycosaminoglycans (GAG) (ii) inhibition of cell proliferation, and (iii) transglutaminase2 (TG2) mediated crosslinking of ps20 to fibronectin, a process implicated in wound healing. We show herein that ps20 preparations contain multiple molecular forms including full-length ps20 (resolving at ≈27 kDa), an exon 3 truncated form (≈22 kDa) that lacks aa113–140, and variable amounts of a putatively cleaved lower MW (≈15–17 kDa) species. Untagged purified ps20 preparations containing a mixture of these forms are biologically active in significantly suppressing prostate cell proliferation. We show that one mechanism by which lower LMW forms of ps20 arise is through cathepsin L (CL) cleavage, and confirm that CL cleaves ps20 at the C-terminus, but this does not inhibit its growth inhibitory function. However, CL cleavage abrogated the interaction between ps20 and solid-phase fibronectin.Therefore, we demonstrate for the first time that LMW forms of ps20 that lack a C-terminal immunogenic epitope can arise through CL cleavage and this cleavage impairs multimerisation and potential capacity to cross-link to ECM, but not the capacity of ps20 to inhibit cell proliferation. We propose that ps20 like other WFDC proteins can become associated with GAGs and the ECM. Furthermore, we suggest post-translational processing and cleavage of ps20 is required to generate functional protein species, and TG2 mediated crosslinking and CL cleavage form components of a ps20 regulatory apparatus.

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Molecular structure, gene expression and functional role of WFDC1 in angiogenesis and cancer

Zhu

Bennett

et al. 2021

Cell Biochemistry & Function

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Whey acidic proteins (WAP) perform a diverse range of important biological functions, including proteinase activity, calcium transport and bacterial growth. The WAP four‐disulphide core domain protein 1 (WFDC1) gene (also called PS20), encodes the 20 kDa prostate stromal protein (ps20), which is a member of the WAP‐type four‐disulphide core domain family of proteins, and exhibits characteristics of serine protease inhibitors, such as elafin and secretory leukocyte protease inhibitor. Molecular structural analysis reveals that ps20 consists of four‐disulphide bonds formed by eight cysteine residues located at the carboxyl terminus of the protein. Wfdc1‐null mice were found to display no overt developmental phenotype, suggesting a dispensable role in organ growth and development. However, WFDC1 was able to mediate endothelial cell migration and pericyte stabilization, which are vital for the formation of functional vascular structures. WFDC1 was also found to be downregulated in cancers and exhibited a regulatory effect on cell proliferation. In addition, it was involved in the modulation of memory T cells during human immunodeficiency virus infection. Gaining a solid understanding of the mechanisms by which WFDC1 regulates tissue homeostasis and disease processes, in a tissue specific manner, will be an important move towards the development of WFDC1/ps20 as potential therapeutic targets.

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Expression of two WFDC1/ps20 isoforms in prostate stromal cells induces paracrine apoptosis through regulation of PTGS2/COX-2

Cited by 7 publications

References 31 publications

Downregulated microRNA-26a modulates prostate cancer cell proliferation and apoptosis by targeting COX-2

Downregulated microRNA-26a modulates prostate cancer cell proliferation and apoptosis by targeting COX-2

Cathepsin-L and transglutaminase dependent processing of ps20: A novel mechanism for ps20 regulation via ECM cross-linking

Molecular structure, gene expression and functional role of WFDC1 in angiogenesis and cancer

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