Limits...
Cathelicidin suppresses colon cancer development by inhibition of cancer associated fibroblasts.

Cheng M, Ho S, Yoo JH, Tran DH, Bakirtzi K, Su B, Tran DH, Kubota Y, Ichikawa R, Koon HW - Clin Exp Gastroenterol (2014)

Bottom Line: Intravenous administration of cathelicidin expressing adeno-associated virus significantly reduced the size of tumors, tumor-derived collagen expression, and tumor-derived fibroblast expression in HT-29-derived subcutaneous tumors in nude mice.Cathelicidin did not directly affect HT-29 cell viability, but did significantly reduce tumor growth factor-β1-induced EMT of colon cancer cells.Cathelicidin disrupted tubulin distribution in colonic fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Center for Inflammatory Bowel Diseases, Division of Digestive Diseases, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.

ABSTRACT

Background: Cathelicidin (LL-37 in humans and mCRAMP in mice) represents a family of endogenous antimicrobial and anti-inflammatory peptides. Cancer-associated fibroblasts can promote the proliferation of colon cancer cells and growth of colon cancer tumors.

Methods: We examined the role of cathelicidin in the development of colon cancer, using subcutaneous human HT-29 colon-cancer-cell-derived tumor model in nude mice and azoxymethane- and dextran sulfate-mediated colon cancer model in C57BL/6 mice. We also determined the indirect antitumoral mechanism of cathelicidin via the inhibition of epithelial-mesenchymal transition (EMT) of colon cancer cells and fibroblast-supported colon cancer cell proliferation.

Results: Intravenous administration of cathelicidin expressing adeno-associated virus significantly reduced the size of tumors, tumor-derived collagen expression, and tumor-derived fibroblast expression in HT-29-derived subcutaneous tumors in nude mice. Enema administration of the mouse cathelicidin peptide significantly reduced the size and number of colonic tumors in azoxymethane- and dextran sulfate-treated mice without inducing apoptosis in tumors and the adjacent normal colonic tissues. Cathelicidin inhibited the collagen expression and vimentin-positive fibroblast expression in colonic tumors. Cathelicidin did not directly affect HT-29 cell viability, but did significantly reduce tumor growth factor-β1-induced EMT of colon cancer cells. Media conditioned by the human colonic CCD-18Co fibroblasts promoted human colon cancer HT-29 cell proliferation. Cathelicidin pretreatment inhibited colon cancer cell proliferation mediated by media conditioned by human colonic CCD-18Co fibroblasts. Cathelicidin disrupted tubulin distribution in colonic fibroblasts. Disruption of tubulin in fibroblasts reduced fibroblast-supported colon cancer cell proliferation.

Conclusion: Cathelicidin effectively inhibits colon cancer development by interfering with EMT and fibroblast-supported colon cancer cell proliferation.

No MeSH data available.


Related in: MedlinePlus

Cathelicidin inhibits EMT and associated cell morphology changes of HT-29 cells. (A and B) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 48 hours. TGF-β1-induced α-SMA and vimentin mRNA expression was significantly inhibited by LL-37. (C) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. TGF-β reduced HT-29 cell colony cell size, and some cells changed into a fibroblast-like spindle shape (see arrows) which was reversed by the co-incubation with LL-37. Results are representative of three independent experiments. (D) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. Protein expression of EMT mediators was determined by Western blot analyses. TGF-β1 increased N-cadherin but not the Slug, Twist1, or E-cadherin protein expression. LL-37 (10 μM) reduced N-cadherin, Slug, and Twist1 but not E-cadherin protein expression in the presence of TGF-β1. Results are representative of two independent experiments.Abbreviations: EMT, epithelial–mesenchymal transition; α-SMA, alpha smooth muscle actin; mRNA, messenger RNA; TFA, trifluoroacetic acid; TGF-β1, tumor growth factor-β1.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4274046&req=5

f6-ceg-8-013: Cathelicidin inhibits EMT and associated cell morphology changes of HT-29 cells. (A and B) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 48 hours. TGF-β1-induced α-SMA and vimentin mRNA expression was significantly inhibited by LL-37. (C) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. TGF-β reduced HT-29 cell colony cell size, and some cells changed into a fibroblast-like spindle shape (see arrows) which was reversed by the co-incubation with LL-37. Results are representative of three independent experiments. (D) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. Protein expression of EMT mediators was determined by Western blot analyses. TGF-β1 increased N-cadherin but not the Slug, Twist1, or E-cadherin protein expression. LL-37 (10 μM) reduced N-cadherin, Slug, and Twist1 but not E-cadherin protein expression in the presence of TGF-β1. Results are representative of two independent experiments.Abbreviations: EMT, epithelial–mesenchymal transition; α-SMA, alpha smooth muscle actin; mRNA, messenger RNA; TFA, trifluoroacetic acid; TGF-β1, tumor growth factor-β1.

Mentions: Fibroblasts and cells with mesenchymal phenotype express vimentin.18,26 TGF-β1 can induce EMT in colon cancer cells with the expression of vimentin and α-SMA.27 Cathelicidin inhibited TGF-β1-induced expression of α-SMA and vimentin in HT-29 colon cancer cells, suggesting the inhibition of transition from an epithelial cell phenotype to a mesenchymal cell phenotype (Figure 6A and B). In addition, exposure of epithelial HT-29 colon cancer cells to TGF-β1 for 48 hours induced EMT as characterized by partial change to spindle fibroblast-like cell shape and loss of colony size (Figure 6C). Co-incubation with human cathelicidin LL-37 (10 μM) prevented TGF-β1-induced change of cell shape and maintained the relatively large HT-29 cell colony size (Figure 6C). In the EMT process, N-cadherin Twist and Slug expressions increased, while E-cadherin expression decreased.26,28 Similarly, exposure of epithelial HT-29 cells to TGF-β1 for 72 hours increased N-cadherin protein expression, indicating mesenchymal phenotype transition (Figure 6D). Co-incubation with LL-37 (10 μM) suppressed TGF-β1-induced N-cadherin expression, indicating the inhibition of EMT (Figure 6D). However, TGF-β1 exposure did not change the basal E-cadherin protein expression, suggesting the preservation of some epithelial cell phenotype (Figure 6D). LL-37 mildly reduced EMT molecule Twist1 and Slug protein expression in the presence of TGF-β1 but did not alter E-cadherin protein expression (Figure 6D). Inhibition of EMT may reduce the availability of mesenchymal cells with fibroblast phenotypes that support colon cancer cell proliferation.


Cathelicidin suppresses colon cancer development by inhibition of cancer associated fibroblasts.

Cheng M, Ho S, Yoo JH, Tran DH, Bakirtzi K, Su B, Tran DH, Kubota Y, Ichikawa R, Koon HW - Clin Exp Gastroenterol (2014)

Cathelicidin inhibits EMT and associated cell morphology changes of HT-29 cells. (A and B) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 48 hours. TGF-β1-induced α-SMA and vimentin mRNA expression was significantly inhibited by LL-37. (C) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. TGF-β reduced HT-29 cell colony cell size, and some cells changed into a fibroblast-like spindle shape (see arrows) which was reversed by the co-incubation with LL-37. Results are representative of three independent experiments. (D) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. Protein expression of EMT mediators was determined by Western blot analyses. TGF-β1 increased N-cadherin but not the Slug, Twist1, or E-cadherin protein expression. LL-37 (10 μM) reduced N-cadherin, Slug, and Twist1 but not E-cadherin protein expression in the presence of TGF-β1. Results are representative of two independent experiments.Abbreviations: EMT, epithelial–mesenchymal transition; α-SMA, alpha smooth muscle actin; mRNA, messenger RNA; TFA, trifluoroacetic acid; TGF-β1, tumor growth factor-β1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4274046&req=5

f6-ceg-8-013: Cathelicidin inhibits EMT and associated cell morphology changes of HT-29 cells. (A and B) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 48 hours. TGF-β1-induced α-SMA and vimentin mRNA expression was significantly inhibited by LL-37. (C) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. TGF-β reduced HT-29 cell colony cell size, and some cells changed into a fibroblast-like spindle shape (see arrows) which was reversed by the co-incubation with LL-37. Results are representative of three independent experiments. (D) HT-29 cells were treated with TGF-β1 (40 ng/mL) with or without LL-37 for 72 hours. Protein expression of EMT mediators was determined by Western blot analyses. TGF-β1 increased N-cadherin but not the Slug, Twist1, or E-cadherin protein expression. LL-37 (10 μM) reduced N-cadherin, Slug, and Twist1 but not E-cadherin protein expression in the presence of TGF-β1. Results are representative of two independent experiments.Abbreviations: EMT, epithelial–mesenchymal transition; α-SMA, alpha smooth muscle actin; mRNA, messenger RNA; TFA, trifluoroacetic acid; TGF-β1, tumor growth factor-β1.
Mentions: Fibroblasts and cells with mesenchymal phenotype express vimentin.18,26 TGF-β1 can induce EMT in colon cancer cells with the expression of vimentin and α-SMA.27 Cathelicidin inhibited TGF-β1-induced expression of α-SMA and vimentin in HT-29 colon cancer cells, suggesting the inhibition of transition from an epithelial cell phenotype to a mesenchymal cell phenotype (Figure 6A and B). In addition, exposure of epithelial HT-29 colon cancer cells to TGF-β1 for 48 hours induced EMT as characterized by partial change to spindle fibroblast-like cell shape and loss of colony size (Figure 6C). Co-incubation with human cathelicidin LL-37 (10 μM) prevented TGF-β1-induced change of cell shape and maintained the relatively large HT-29 cell colony size (Figure 6C). In the EMT process, N-cadherin Twist and Slug expressions increased, while E-cadherin expression decreased.26,28 Similarly, exposure of epithelial HT-29 cells to TGF-β1 for 72 hours increased N-cadherin protein expression, indicating mesenchymal phenotype transition (Figure 6D). Co-incubation with LL-37 (10 μM) suppressed TGF-β1-induced N-cadherin expression, indicating the inhibition of EMT (Figure 6D). However, TGF-β1 exposure did not change the basal E-cadherin protein expression, suggesting the preservation of some epithelial cell phenotype (Figure 6D). LL-37 mildly reduced EMT molecule Twist1 and Slug protein expression in the presence of TGF-β1 but did not alter E-cadherin protein expression (Figure 6D). Inhibition of EMT may reduce the availability of mesenchymal cells with fibroblast phenotypes that support colon cancer cell proliferation.

Bottom Line: Intravenous administration of cathelicidin expressing adeno-associated virus significantly reduced the size of tumors, tumor-derived collagen expression, and tumor-derived fibroblast expression in HT-29-derived subcutaneous tumors in nude mice.Cathelicidin did not directly affect HT-29 cell viability, but did significantly reduce tumor growth factor-β1-induced EMT of colon cancer cells.Cathelicidin disrupted tubulin distribution in colonic fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Center for Inflammatory Bowel Diseases, Division of Digestive Diseases, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.

ABSTRACT

Background: Cathelicidin (LL-37 in humans and mCRAMP in mice) represents a family of endogenous antimicrobial and anti-inflammatory peptides. Cancer-associated fibroblasts can promote the proliferation of colon cancer cells and growth of colon cancer tumors.

Methods: We examined the role of cathelicidin in the development of colon cancer, using subcutaneous human HT-29 colon-cancer-cell-derived tumor model in nude mice and azoxymethane- and dextran sulfate-mediated colon cancer model in C57BL/6 mice. We also determined the indirect antitumoral mechanism of cathelicidin via the inhibition of epithelial-mesenchymal transition (EMT) of colon cancer cells and fibroblast-supported colon cancer cell proliferation.

Results: Intravenous administration of cathelicidin expressing adeno-associated virus significantly reduced the size of tumors, tumor-derived collagen expression, and tumor-derived fibroblast expression in HT-29-derived subcutaneous tumors in nude mice. Enema administration of the mouse cathelicidin peptide significantly reduced the size and number of colonic tumors in azoxymethane- and dextran sulfate-treated mice without inducing apoptosis in tumors and the adjacent normal colonic tissues. Cathelicidin inhibited the collagen expression and vimentin-positive fibroblast expression in colonic tumors. Cathelicidin did not directly affect HT-29 cell viability, but did significantly reduce tumor growth factor-β1-induced EMT of colon cancer cells. Media conditioned by the human colonic CCD-18Co fibroblasts promoted human colon cancer HT-29 cell proliferation. Cathelicidin pretreatment inhibited colon cancer cell proliferation mediated by media conditioned by human colonic CCD-18Co fibroblasts. Cathelicidin disrupted tubulin distribution in colonic fibroblasts. Disruption of tubulin in fibroblasts reduced fibroblast-supported colon cancer cell proliferation.

Conclusion: Cathelicidin effectively inhibits colon cancer development by interfering with EMT and fibroblast-supported colon cancer cell proliferation.

No MeSH data available.


Related in: MedlinePlus