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Anti-IL-20 Monoclonal Antibody Suppresses Prostate Cancer Growth and Bone Osteolysis in Murine Models.

Hsu YH, Wu CY, Hsing CH, Lai WT, Wu LW, Chang MS - PLoS ONE (2015)

Bottom Line: IL-20 is associated with tumor promotion in the pathogenesis of oral, bladder, and breast cancer.In vivo, 7E reduced tumor growth, suppressed tumor-mediated osteolysis, and protected bone mineral density after intratibial injection of prostate cancer cells.We conclude that IL-20 is involved in the cell migration, colony formation, and tumor-induced osteolysis of prostate cancer.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

ABSTRACT
Interleukin (IL)-20 is a proinflammatory cytokine in the IL-10 family. IL-20 is associated with tumor promotion in the pathogenesis of oral, bladder, and breast cancer. However, little is known about the role of IL-20 in prostate cancer. We hypothesize that IL-20 promotes the growth of prostate cancer cells. Immunohistochemical staining showed that IL-20 and its receptors were expressed in human PC-3 and LNCaP prostate cancer cell lines and in prostate tumor tissue from 40 patients. In vitro, IL-20 upregulated N-cadherin, STAT3, vimentin, fibronectin, RANKL, cathepsin G, and cathepsin K, and increased the migration and colony formation of prostate cancer cells via activated p38, ERK1/2, AKT, and NF-κB signals in PC-3 cells. We investigated the effects of anti-IL-20 monoclonal antibody 7E on prostate tumor growth in vivo using SCID mouse subcutaneous and intratibial xenograft tumor models. In vivo, 7E reduced tumor growth, suppressed tumor-mediated osteolysis, and protected bone mineral density after intratibial injection of prostate cancer cells. We conclude that IL-20 is involved in the cell migration, colony formation, and tumor-induced osteolysis of prostate cancer. Therefore, IL-20 might be a novel target for treating prostate cancer.

No MeSH data available.


Related in: MedlinePlus

RANKL, cathepsin G, and cathepsin K transcripts were upregulated and sRANKL protein production was promoted in IL-20-treated PC–3 cells.(A-C) PC–3 cells were treated with hIL–20 (200 ng/ml), 7E (2 μg/ml), mIgG (2 μg/ml), hIL–20 plus 7E, or hIL–20 plus mIgG for 6 hours, and the expression levels of RANKL, cathepsin G, and cathepsin K were analyzed using RT-qPCR with specific primers. *p < 0.05, **p < 0.01 versus untreated control, #p < 0.05 versus the hIL–20 group. Data are representative of 3 independent experiments, each done in triplicate. (D) PC–3 cells were incubated with hIL–20 (100, 200, or 400 ng/ml) for 48 hours, the culture medium was collected and then the concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls. Data are representative of 3 independent experiments, each done in triplicate. (E) PC–3 cells were preincubated with 1 μM of cathepsin G-specific inhibitor (TPCK) for 1 hour and then treated with hIL–20 (400 ng/ml) for 72 hours. The vehicle control was 0.0175% EtOH in culture medium. The concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls, #p < 0.05 versus the hIL–20 plus EtOH vehicle controls. Data are the means ± SD of three independent experiments, each done in triplicate. RT-qPCR, real-time quantitative polymerase chain reaction; sRANKL, soluble RANKL; EtOH, ethanol.
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pone.0139871.g006: RANKL, cathepsin G, and cathepsin K transcripts were upregulated and sRANKL protein production was promoted in IL-20-treated PC–3 cells.(A-C) PC–3 cells were treated with hIL–20 (200 ng/ml), 7E (2 μg/ml), mIgG (2 μg/ml), hIL–20 plus 7E, or hIL–20 plus mIgG for 6 hours, and the expression levels of RANKL, cathepsin G, and cathepsin K were analyzed using RT-qPCR with specific primers. *p < 0.05, **p < 0.01 versus untreated control, #p < 0.05 versus the hIL–20 group. Data are representative of 3 independent experiments, each done in triplicate. (D) PC–3 cells were incubated with hIL–20 (100, 200, or 400 ng/ml) for 48 hours, the culture medium was collected and then the concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls. Data are representative of 3 independent experiments, each done in triplicate. (E) PC–3 cells were preincubated with 1 μM of cathepsin G-specific inhibitor (TPCK) for 1 hour and then treated with hIL–20 (400 ng/ml) for 72 hours. The vehicle control was 0.0175% EtOH in culture medium. The concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls, #p < 0.05 versus the hIL–20 plus EtOH vehicle controls. Data are the means ± SD of three independent experiments, each done in triplicate. RT-qPCR, real-time quantitative polymerase chain reaction; sRANKL, soluble RANKL; EtOH, ethanol.

Mentions: To test whether IL–20 regulates cathepsins and RANKL in prostate cancer, PC–3 cells were treated with IL–20 for 6 hours. An RT-qPCR gene transcript analysis showed upregulated RANKL, cathepsin G and cathepsin K expression in IL-20-treated PC–3 cells, the activity of which was neutralized by 7E (Fig 6A, 6B and 6C). Moreover, an ELISA assay showed a significant (p < 0.05) increase in sRANKL expression in IL-20-treated PC–3 cells (Fig 6D). Therefore, we hypothesized that IL-20-treated PC–3 cells produce cathepsin G and subsequently cleave RANKL to generate sRANKL, which further promotes osteoclast activation in bone microenvironment. To confirm that the cleavage of RANKL was cathepsin G-dependent, we used a specific cathepsin G inhibitor (TPCK, 1 μM) to block the protease activity of cathepsin G. The result confirmed our hypothesis (Fig 6E).


Anti-IL-20 Monoclonal Antibody Suppresses Prostate Cancer Growth and Bone Osteolysis in Murine Models.

Hsu YH, Wu CY, Hsing CH, Lai WT, Wu LW, Chang MS - PLoS ONE (2015)

RANKL, cathepsin G, and cathepsin K transcripts were upregulated and sRANKL protein production was promoted in IL-20-treated PC–3 cells.(A-C) PC–3 cells were treated with hIL–20 (200 ng/ml), 7E (2 μg/ml), mIgG (2 μg/ml), hIL–20 plus 7E, or hIL–20 plus mIgG for 6 hours, and the expression levels of RANKL, cathepsin G, and cathepsin K were analyzed using RT-qPCR with specific primers. *p < 0.05, **p < 0.01 versus untreated control, #p < 0.05 versus the hIL–20 group. Data are representative of 3 independent experiments, each done in triplicate. (D) PC–3 cells were incubated with hIL–20 (100, 200, or 400 ng/ml) for 48 hours, the culture medium was collected and then the concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls. Data are representative of 3 independent experiments, each done in triplicate. (E) PC–3 cells were preincubated with 1 μM of cathepsin G-specific inhibitor (TPCK) for 1 hour and then treated with hIL–20 (400 ng/ml) for 72 hours. The vehicle control was 0.0175% EtOH in culture medium. The concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls, #p < 0.05 versus the hIL–20 plus EtOH vehicle controls. Data are the means ± SD of three independent experiments, each done in triplicate. RT-qPCR, real-time quantitative polymerase chain reaction; sRANKL, soluble RANKL; EtOH, ethanol.
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pone.0139871.g006: RANKL, cathepsin G, and cathepsin K transcripts were upregulated and sRANKL protein production was promoted in IL-20-treated PC–3 cells.(A-C) PC–3 cells were treated with hIL–20 (200 ng/ml), 7E (2 μg/ml), mIgG (2 μg/ml), hIL–20 plus 7E, or hIL–20 plus mIgG for 6 hours, and the expression levels of RANKL, cathepsin G, and cathepsin K were analyzed using RT-qPCR with specific primers. *p < 0.05, **p < 0.01 versus untreated control, #p < 0.05 versus the hIL–20 group. Data are representative of 3 independent experiments, each done in triplicate. (D) PC–3 cells were incubated with hIL–20 (100, 200, or 400 ng/ml) for 48 hours, the culture medium was collected and then the concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls. Data are representative of 3 independent experiments, each done in triplicate. (E) PC–3 cells were preincubated with 1 μM of cathepsin G-specific inhibitor (TPCK) for 1 hour and then treated with hIL–20 (400 ng/ml) for 72 hours. The vehicle control was 0.0175% EtOH in culture medium. The concentration of sRANKL was determined using ELISA. *p < 0.05 versus untreated controls, #p < 0.05 versus the hIL–20 plus EtOH vehicle controls. Data are the means ± SD of three independent experiments, each done in triplicate. RT-qPCR, real-time quantitative polymerase chain reaction; sRANKL, soluble RANKL; EtOH, ethanol.
Mentions: To test whether IL–20 regulates cathepsins and RANKL in prostate cancer, PC–3 cells were treated with IL–20 for 6 hours. An RT-qPCR gene transcript analysis showed upregulated RANKL, cathepsin G and cathepsin K expression in IL-20-treated PC–3 cells, the activity of which was neutralized by 7E (Fig 6A, 6B and 6C). Moreover, an ELISA assay showed a significant (p < 0.05) increase in sRANKL expression in IL-20-treated PC–3 cells (Fig 6D). Therefore, we hypothesized that IL-20-treated PC–3 cells produce cathepsin G and subsequently cleave RANKL to generate sRANKL, which further promotes osteoclast activation in bone microenvironment. To confirm that the cleavage of RANKL was cathepsin G-dependent, we used a specific cathepsin G inhibitor (TPCK, 1 μM) to block the protease activity of cathepsin G. The result confirmed our hypothesis (Fig 6E).

Bottom Line: IL-20 is associated with tumor promotion in the pathogenesis of oral, bladder, and breast cancer.In vivo, 7E reduced tumor growth, suppressed tumor-mediated osteolysis, and protected bone mineral density after intratibial injection of prostate cancer cells.We conclude that IL-20 is involved in the cell migration, colony formation, and tumor-induced osteolysis of prostate cancer.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

ABSTRACT
Interleukin (IL)-20 is a proinflammatory cytokine in the IL-10 family. IL-20 is associated with tumor promotion in the pathogenesis of oral, bladder, and breast cancer. However, little is known about the role of IL-20 in prostate cancer. We hypothesize that IL-20 promotes the growth of prostate cancer cells. Immunohistochemical staining showed that IL-20 and its receptors were expressed in human PC-3 and LNCaP prostate cancer cell lines and in prostate tumor tissue from 40 patients. In vitro, IL-20 upregulated N-cadherin, STAT3, vimentin, fibronectin, RANKL, cathepsin G, and cathepsin K, and increased the migration and colony formation of prostate cancer cells via activated p38, ERK1/2, AKT, and NF-κB signals in PC-3 cells. We investigated the effects of anti-IL-20 monoclonal antibody 7E on prostate tumor growth in vivo using SCID mouse subcutaneous and intratibial xenograft tumor models. In vivo, 7E reduced tumor growth, suppressed tumor-mediated osteolysis, and protected bone mineral density after intratibial injection of prostate cancer cells. We conclude that IL-20 is involved in the cell migration, colony formation, and tumor-induced osteolysis of prostate cancer. Therefore, IL-20 might be a novel target for treating prostate cancer.

No MeSH data available.


Related in: MedlinePlus