In vitro treatment of carcinoma cell lines with pancreatic (pro)enzymes suppresses the EMT programme and promotes cell differentiation
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Previous research has suggested a putative utility of pancreatic (pro)enzymes in cancer treatment. The aim of the present study was to investigate the in vitro effects of a mixture of two pancreatic pro-enzymes, i.e., Chymotrypsinogen and Trypsinogen, and the enzyme Amylase on three human cancer cell lines, i.e., OE33 (derived from an oesophageal carcinoma), Panc1 (derived from a pancreatic carcinoma) and Caco-2 (derived from a colon carcinoma).
After treatment of the three cancer cell lines with different doses of the (pro)enzymes for up to 7 days, we observed (i) growth inhibition in a dose-dependent manner, (ii) enhanced expression of β-catenin and E-cadherin and decreased expression of several epithelial-mesenchymal transition (EMT)-associated genes, such as Vimentin, Snail and Slug, (iii) differentiation of Caco-2 cells, including the appearance of cell-specific differentiated structures such as microvilli and tight junctions, the acquisition of a more regular polygonal morphology, and an increased expression of the intestinal differentiation markers alkaline phosphatase and cytokeratin 8, and (iv) differentiation of Panc1 cells, including the formation of cell aggregates, an increment on lamellar bodies and an increased expression of the pancreatic differentiation markers glucagon and insulin.
Our results show that the treatment of three different human cancer cell lines with pancreatic (pro)enzymes results in an enhancement of cell adhesion, an attenuation of several EMT-associated markers, and an increase in the expression of several differentiation-associated markers, suggesting the acquisition of a less malignant phenotype and a decrease in proliferative capacity due to lineage-specific cellular differentiation.
KeywordsCell adhesion Cell differentiation Pancreatic (pro)enzymes Epithelial-mesenchymal transition Cancer treatment
Macarena Peran was supported by a grant from Jaen University, Spain (“Plan de Apoyo a la Investigación, Desarrollo Tecnológico e Innovación. IV. Programa de Ayudas a los Investigadores”). Work in the laboratory of Bath University was funded by Propanc Pty Ltd. Work in the laboratory of J.A. Marchal at Granada University was funded by the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria, FEDER, grant number PI10/02295).
Conflict of interest
Dr Julian Kenyon is the Founder and Scientific Director of Propanc Pty Ltd and owns stock in the company. Propanc Pty Ltd and the University of Bath have filed a joint patent from the scientific research undertaken in this report
- 7.J. Beuth, B. Ost, A. Pakdaman, E. Rethfeldt, P.R. Bock, J. Hanisch et al., Impact of complementary oral enzyme application on the postoperative treatment results of breast cancer patients–results of an epidemiological multicentre retrolective cohort study. Cancer Chemother. Pharmacol. 47, 45–54 (2001)CrossRefGoogle Scholar
- 12.R. Kemler, From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. Trends. Genet. 93, 17–21 (1993)Google Scholar
- 30.M. Pinto, S. Robineleon, M.D. Appay, M. Kedinger, N. Triadou, E. Dussaulx et al., Enterocyte-like differentiation and polarization of the human-colon carcinoma cell-line caco-2 in culture. Biol. Cell 47, 323–330 (1983)Google Scholar
- 34.J.P. Brunet, N. Jourdan, J. Cotte-Laffitte, C. Linxe, M. Geniteau-Legendre, A. Servin et al., Rotavirus infection induces cytoskeleton disorganization in human intestinal epithelial cells: implication of an increase in intracellular calcium concentration. J. Virol. 74, 10801–6 (2000)PubMedCrossRefGoogle Scholar
- 35.R.J. Guang, J.L. Ford, Y.N. Fu, Y.Z. Li, L.M. Shaw, A.B. Pardee, Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. Cancer Res. 60, 749–755 (2000)Google Scholar
- 39.Y. Wu, J. Li, S. Saleem, S.P. Yee, A.A. Hardikar, R. Wang, c-Kit and stem cell factor regulate PANC-1 cell differentiation into insulin- and glucagon-producing cells. Lab. Invest. 90(1373–84) (2010)Google Scholar
- 40.J.A. Marchal, J. Prados, J. Campos, F. González, C. Melguizo, C. Velez et al., Therapeutic potential of differentiation in cancer and normal stem cells, in New Cell Differentiation Research Topics, ed. by H. Saitama (Nova Science Publisher, Inc, New York, 2008), pp. 7–77Google Scholar
- 43.W. Feng, R. Orlandi, N. Zhao, M.L. Carcangiu, E. Tagliabue, J. Xu et al., Tumor suppressor genes are frequently methylated in lymph node metastases of breast cancers. B.M.C. Cancer 10(378–88) (2010)Google Scholar
- 48.X. Tian, Z. Liu, B. Niu, J. Zhang, T.K. Tan, S.R. Lee et al., E-cadherin/beta-catenin complex and the epithelial barrier. J. Biomed. Biotechnol. 2011, 1–6 (2011)Google Scholar
- 49.J.A. Marchal, M.C. Nunez, A. Aranega, M.A. Gallo, A. Espinosa, J.M. Campos, Acyclonucleosides, modified seco-nucleosides, and salicyl- or catechol-derived acyclic 5-fluorouracil O, N-acetals: antiproliferative activities, cellular differentiation and apoptosis. Curr. Med. Chem. 16, 1166–83 (2009)PubMedCrossRefGoogle Scholar