title: Colon cancer cell-derived 12(S)-HETE induces the retraction of cancer-associated fibroblast via MLC2, RHO/ROCK and Ca2+signalling.
abstract: Retraction of mesenchymal stromal cells supports the invasion of colorectal cancer cells (CRC) into the adjacent compartment. CRC-secreted 12(S)-HETE enhances the retraction of cancer-associated fibroblasts (CAFs) and therefore, 12(S)-HETE may enforce invasivity of CRC. Understanding the mechanisms of metastatic CRC is crucial for successful intervention. Therefore, we studied pro-invasive contributions of stromal cells in physiologically relevant three-dimensional in vitro assays consisting of CRC spheroids, CAFs, extracellular matrix and endothelial cells, as well as in reductionist models. In order to elucidate how CAFs support CRC invasion, tumour spheroid-induced CAF retraction and free intracellular Ca2+levels were measured and pharmacological- or siRNA-based inhibition of selected signalling cascades was performed. CRC spheroids caused the retraction of CAFs, generating entry gates in the adjacent surrogate stroma. The responsible trigger factor 12(S)-HETE provoked a signal, which was transduced by PLC, IP3, free intracellular Ca2+, Ca2+-calmodulin-kinase-II, RHO/ROCK and MYLK which led to the activation of myosin light chain 2, and subsequent CAF mobility. RHO activity was observed downstream as well as upstream of Ca2+release. Thus, Ca2+signalling served as central signal amplifier. Treatment with the FDA-approved drugs carbamazepine, cinnarizine, nifedipine and bepridil HCl, which reportedly interfere with cellular calcium availability, inhibited CAF-retraction. The elucidation of signalling pathways and identification of approved inhibitory drugs warrant development of intervention strategies targeting tumour-stroma interaction.
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, 1090, Vienna, Austria.
authors.forename: Serena
authors.initials: S
authors.lastname: Stadler
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Department for Clinical Pharmacy and Diagnostics, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Chi Huu
authors.initials: CH
authors.lastname: Nguyen
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Helga
authors.initials: H
authors.lastname: Schachner
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Daniela
authors.initials: D
authors.lastname: Milovanovic
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, 1090, Vienna, Austria.|Department of Medicine I, Comprehensive Cancer Centre, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Silvio
authors.initials: S
authors.lastname: Holzner
authors.affiliation: Department for Clinical Pharmacy and Diagnostics, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Brenner
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Department of Pharmacognosy, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Julia
authors.initials: J
authors.lastname: Eichsteininger
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, 1090, Vienna, Austria.
authors.forename: Mira
authors.initials: M
authors.lastname: Stadler
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, 1090, Vienna, Austria.|Department of Medicine I, Comprehensive Cancer Centre, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Daniel
authors.initials: D
authors.lastname: Senfter
authors.affiliation: Department of Pharmacognosy, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Liselotte
authors.initials: L
authors.lastname: Krenn
authors.affiliation: Neuromuscular Research Department, Centre of Anatomy and Cell Biology, Medical University of Vienna, Waehringer Strasse 13, 1090, Vienna, Austria.
authors.forename: Wolfgang M
authors.initials: WM
authors.lastname: Schmidt
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Nicole
authors.initials: N
authors.lastname: Huttary
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Sigurd
authors.initials: S
authors.lastname: Krieger
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Oskar
authors.initials: O
authors.lastname: Koperek
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Bago-Horvath
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Konstantin Alexander
authors.initials: KA
authors.lastname: Brendel
authors.affiliation: Department of Medicine I, Institute of Cancer Research and Comprehensive Cancer Centre, Medical University of Vienna, Borschkegasse 8a, 1090, Vienna, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Marian
authors.affiliation: Department of Radiation Oncology and Experimental Cancer Research, Ghent University, De Pintelaan 185, 9000, Ghent, Belgium.
authors.forename: Oliver
authors.initials: O
authors.lastname: de Wever
authors.affiliation: Department of Medicine I, Comprehensive Cancer Centre, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Mader
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria.|Department for Clinical Pharmacy and Diagnostics, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Giessrigl
authors.affiliation: Department for Clinical Pharmacy and Diagnostics, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Walter
authors.initials: W
authors.lastname: Jäger
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Waehringer Strasse 10, 1090, Vienna, Austria.
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Waehringer Guertel 18-20, 1090, Vienna, Austria. georg.krupitza@meduniwien.ac.at.
authors.forename: Georg
authors.initials: G
authors.lastname: Krupitza
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D000072645:Cancer-Associated Fibroblasts / Q000378:metabolism / Q000473:pathology*; D024722:Cardiac Myosins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D003106:Colon / Q000378:metabolism / Q000473:pathology*; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D018994:Myosin Light Chains / Q000378:metabolism; D009361:Neoplasm Invasiveness / Q000473:pathology; D012007:Rectum / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D018994:Myosin Light Chains; C107184:myosin light chain 2; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D054460:rho-Associated Kinases; D024722:Cardiac Myosins; D002118:Calcium
keywords: 3D invasion model; Arachidonic acid metabolite; ECM; Signal transduction; Tumour progression