Phospholipase D activation mediates growth and migration of colon cancer cells interacting with cancer-associated fibroblasts

Salem Majdop; Yehuda Skornick; Shmuel Avital; Liron Berkovich

doi:10.14715/cmb/2018.64.14.14

Issue

Vol. 64 No. 14: Issue 14

Issue Published : December 3, 2018

The undersigned hereby assign all rights, included but not limited to copyright, for this manuscript to CMB Association upon its submission for consideration to publication on Cellular and Molecular Biology. The rights assigned include, but are not limited to, the sole and exclusive rights to license, sell, subsequently assign, derive, distribute, display and reproduce this manuscript, in whole or in part, in any format, electronic or otherwise, including those in existence at the time this agreement was signed. The authors hereby warrant that they have not granted or assigned, and shall not grant or assign, the aforementioned rights to any other person, firm, organization, or other entity. All rights are automatically restored to authors if this manuscript is not accepted for publication.

Phospholipase D activation mediates growth and migration of colon cancer cells interacting with cancer-associated fibroblasts

https://doi.org/10.14715/cmb/2018.64.14.14

Salem Majdop

Department of Surgery B and Surgery B Cancer Research Laboratory, Meir Medical Center, 59 Tshernichovsky, Kfar Saba, 44281 Israel

Yehuda Skornick

Department of Surgery B and Surgery B Cancer Research Laboratory, Meir Medical Center, 59 Tshernichovsky, Kfar Saba, 44281 Israel

Shmuel Avital

Department of Surgery B and Surgery B Cancer Research Laboratory, Meir Medical Center, 59 Tshernichovsky, Kfar Saba, 44281 Israel

Liron Berkovich

Department of Surgery B and Surgery B Cancer Research Laboratory, Meir Medical Center, 59 Tshernichovsky, Kfar Saba, 44281 Israel

Corresponding Author(s) : Liron Berkovich

liron.berkovich@gmail.com

Cellular and Molecular Biology, Vol. 64 No. 14: Issue 14
Article Published : November 30, 2018

Abstract

Cancer-associated fibroblasts of the stroma play a major role in tumor promoting processes. In this study we evaluated the significance of Phospholipase D (PLD) enzyme activity in promoting human colon cancer malignant potency when interacting with proximal colonic fibroblasts. Human colon cancer cell lines SW480 and HCT116, and colonic fibroblasts CCD-18Co were used as an in vitro model. PLD's activity was measured in resting cancer cells and after culturing with fibroblasts and cancer-associated fibroblasts (CAFs) conditioned medium. The viability and migration level of the cancer cells alone and after co-culturing with fibroblast or CAFs conditioned medium were evaluated, with and without adding a PLD inhibitor. Exposure of colon cancer cells to CAFs conditioned medium significantly increased the level of PLD activity in the cancer cells (p<0.0001). Exposure of colon cancer to resting and activated fibroblast conditioned medium significantly enhanced the number of viable cancer cells as well as its migration level measured following 24 and 48 hours. Adding a PLD inhibitor significantly reduced the elevation of cell viability and migration of the colon cancer cells exposed to fibroblasts conditioned medium (p<0.005). In this in vitro model, inhibition of PLD significantly decreased proliferation and migration levels of colon cancer cells generated by stromal fibroblasts. This provides evidence that the PLD signaling pathway is directly involved in stroma-cancer interactions in the colon, thereby promoting cancer progression. Further research is needed in order to evaluate PLD as a target in colon cancer prevention or therapy.

Keywords

Phospholipase D Colon cancer Stroma Cancer-associated fibroblasts.

Majdop, S., Skornick, Y., Avital, S., & Berkovich, L. (2018). Phospholipase D activation mediates growth and migration of colon cancer cells interacting with cancer-associated fibroblasts. Cellular and Molecular Biology, 64(14), 84–88. https://doi.org/10.14715/cmb/2018.64.14.14

Download Citation

References

Weinberg RA. The Biology of Cancer, Garland Science. 2007.
Pietras K, Ostman A. Hallmarks of cancer: interactions with the tumor stroma. Exp Cell Res. 2010;316:1324-31.
Räsänen K, Vaheri A. Activation of fibroblasts in cancer stroma. Exp Cell Res. 2010;316:2713-22.
Huijbers A, Tollenaar RA, Pelt GW, Zeestraten EC, Dutton S, McConkey CC, et al. The proportion of tumor-stroma as a strong prognosticator for stage II and III colon cancer patients: validation in the VICTOR trial. Ann Oncol. 2013;24:179-185.
West NP, Dattani M, McShane P, Hutchins G, Grabsch J, Mueller W, et al. The proportion of tumour cells is an independent predictor for survival in colorectal cancer patients. Br J Cancer. 2010;102:1519-23.
Tommelein J, Verset L, Boterberg T, Demetter P, Bracke M, De Wever O. Cancer-associated fibroblasts connect metastasis promoting communication in colorectal cancer. Front Oncol. 2015;5:63.
Herrera M, Islam AB, Herrera A, Martín P, García V, Silva J, et al. Functional heterogeneity of cancer-associated fibroblasts from human colon tumors shows specific prognostic gene expression signature. Clin Cancer Res. 2013;19:5914-26.
Huang P, Frohman MA. The potential for phospholipase D as a new therapeutic target. Expert Opin Ther Targets. 2007;11:707-16.
Su W, Chen Q, Frohman MA.Targeting phospholipase D with small-molecule inhibitors as a potential therapeutic approach for cancer metastasis. Future Oncol. 2009;5(9):1477-86.
Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protocols. 2007;2:329-33.
Feoktistova M, Geserick P, Leverkus M. CrystalViolet Assay for Determining Viability of Cultured Cells. Cold Spring Harb Protoc. 2016; 1;2016(4):pdb.prot087379.
Gomez-Cambronero, J. Phosphatidic acid, phospholipase D and tumorigenesis. Advances in biological regulation. 2014;54:197-206.
Saito M, Iwadate M, Higashimoto M, Ono K, Takebayashi Y, Takenoshita S. Expression of phospholipase D2 in human colorectal carcinoma. Oncol Rep 2007; 18: 1329–1334.
Henkels KM, Boivin GP, Dudley ES, Berberich SJ, Gomez-Cambronero J. Phospholipase D (PLD) drives cell invasion, tumor growth and metastasis in a human breast cancer xenograph model. Oncogene. 2013;32:5551-62.
Baan B, Dihal AA, Hoff E, Bos CL, Voorneveld PW, Koelink PJ, et al. 5-aminosalicylic acid inhibits cell cycle progression in a phospholipase D dependent manner in colorectal cancer. Gut 2012;61:1708–15.

References

Weinberg RA. The Biology of Cancer, Garland Science. 2007.

Pietras K, Ostman A. Hallmarks of cancer: interactions with the tumor stroma. Exp Cell Res. 2010;316:1324-31.

Räsänen K, Vaheri A. Activation of fibroblasts in cancer stroma. Exp Cell Res. 2010;316:2713-22.

Huijbers A, Tollenaar RA, Pelt GW, Zeestraten EC, Dutton S, McConkey CC, et al. The proportion of tumor-stroma as a strong prognosticator for stage II and III colon cancer patients: validation in the VICTOR trial. Ann Oncol. 2013;24:179-185.

West NP, Dattani M, McShane P, Hutchins G, Grabsch J, Mueller W, et al. The proportion of tumour cells is an independent predictor for survival in colorectal cancer patients. Br J Cancer. 2010;102:1519-23.

Tommelein J, Verset L, Boterberg T, Demetter P, Bracke M, De Wever O. Cancer-associated fibroblasts connect metastasis promoting communication in colorectal cancer. Front Oncol. 2015;5:63.

Herrera M, Islam AB, Herrera A, Martín P, García V, Silva J, et al. Functional heterogeneity of cancer-associated fibroblasts from human colon tumors shows specific prognostic gene expression signature. Clin Cancer Res. 2013;19:5914-26.

Huang P, Frohman MA. The potential for phospholipase D as a new therapeutic target. Expert Opin Ther Targets. 2007;11:707-16.

Su W, Chen Q, Frohman MA.Targeting phospholipase D with small-molecule inhibitors as a potential therapeutic approach for cancer metastasis. Future Oncol. 2009;5(9):1477-86.

Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protocols. 2007;2:329-33.

Feoktistova M, Geserick P, Leverkus M. CrystalViolet Assay for Determining Viability of Cultured Cells. Cold Spring Harb Protoc. 2016; 1;2016(4):pdb.prot087379.

Gomez-Cambronero, J. Phosphatidic acid, phospholipase D and tumorigenesis. Advances in biological regulation. 2014;54:197-206.

Saito M, Iwadate M, Higashimoto M, Ono K, Takebayashi Y, Takenoshita S. Expression of phospholipase D2 in human colorectal carcinoma. Oncol Rep 2007; 18: 1329–1334.

Henkels KM, Boivin GP, Dudley ES, Berberich SJ, Gomez-Cambronero J. Phospholipase D (PLD) drives cell invasion, tumor growth and metastasis in a human breast cancer xenograph model. Oncogene. 2013;32:5551-62.

Baan B, Dihal AA, Hoff E, Bos CL, Voorneveld PW, Koelink PJ, et al. 5-aminosalicylic acid inhibits cell cycle progression in a phospholipase D dependent manner in colorectal cancer. Gut 2012;61:1708–15.

Author biographies is not available.