title: Evaluation of pancreatic cancer cell migration with multiple parameters in vitro by using an optical real-time cell mobility assay device.
abstract: Migration of cancer cell correlates with distant metastasis and local invasion, which are good targets for cancer treatment. An optically accessible device "TAXIScan" was developed, which provides considerably more information regarding the cellular dynamics and less quantity of samples than do the existing methods. Here, we report the establishment of a system to analyze the nature of pancreatic cancer cells using TAXIScan and we evaluated lysophosphatidic acid (LPA)-elicited pancreatic cell migration.
  
  
  
  Pancreatic cancer cell lines, BxPC3, PANC-1, AsPC1, and MIAPaCa-2, were analyzed for adhesion as well as migration towards LPA by TAXIScan using parameters such as velocity and directionality or for the number of migrated cells by the Boyden chamber methods. To confirm that the migration was initiated by LPA, the expression of LPA receptors and activation of intracellular signal transductions were examined by quantitative reverse transcriptase polymerase reaction and western blotting.
  
  
  
  Scaffold coating was necessary for the adhesion of pancreatic cancer cells, and collagen I and Matrigel were found to be good scaffolds. BxPC3 and PANC-1 cells clearly migrated towards the concentration gradient formed by injecting 1 μL LPA, which was abrogated by pre-treatment with LPA inhibitor, Ki16425 (IC50for the directionality ≈ 1.86 μM). The LPA dependent migration was further confirmed by mRNA and protein expression of LPA receptors as well as phosphorylation of signaling molecules. LPA1mRNA was highest among the 6 receptors, and LPA1, LPA2and LPA3proteins were detected in BxPC3 and PANC-1 cells. Phosphorylation of Akt (Thr308 and Ser473) and p42/44MAPK in BxPC3 and PANC-1 cells was observed after LPA stimulation, which was clearly inhibited by pre-treatment with a compound Ki16425.
  
  
  
  We established a novel pancreatic cancer cell migration assay system using TAXIScan. This assay device provides multiple information on migrating cells simultaneously, such as their morphology, directionality, and velocity, with a small volume of sample and can be a powerful tool for analyzing the nature of cancer cells and for identifying new factors that affect cell functions.
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan. akiray@med.kawasaki-m.ac.jp.
authors.forename: Akira
authors.identifier: 0000-0001-9205-6922
authors.initials: A
authors.lastname: Yamauchi
authors.affiliation: Department of Clinical Oncology, Kawasaki Medical School, Okayama, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamamura
authors.affiliation: Department of Molecular and Developmental Biology, Kawasaki Medical School, Okayama, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Katase
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan.
authors.forename: Masumi
authors.initials: M
authors.lastname: Itadani
authors.affiliation: Department of Clinical Oncology, Kawasaki Medical School, Okayama, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Okada
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan.
authors.forename: Kayoko
authors.initials: K
authors.lastname: Kobiki
authors.affiliation: Department of Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Department of Clinical Oncology, Kawasaki Medical School, Okayama, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Yamaguchi
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan.
authors.forename: Futoshi
authors.initials: F
authors.lastname: Kuribayashi
mesh_terms: D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D008246:Lysophospholipids / Q000494:pharmacology*; D055096:Optical Devices / Q000706:statistics & numerical data*; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000473:pathology*; D049368:Receptors, Lysophosphatidic Acid / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured
chemical_list: D008246:Lysophospholipids; D049368:Receptors, Lysophosphatidic Acid; C032881:lysophosphatidic acid
keywords: Chemotaxis; Inhibitor; Lipid mediator; Metastasis; Migration; TAXIScan