Evaluation of pancreatic cancer cell migration with multiple parameters in vitro by using an optical real-time cell mobility assay device

BMC Cancer

Research Article

Akira Yamauchi¹  Kayoko Kobiki¹  Futoshi Kuribayashi¹  Masumi Itadani¹  Naoko Okada²  Masahiro Yamamura²  Yoshiyuki Yamaguchi²  Naoki Katase³  Masafumi Nakamura⁴ 

[1] Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki, 701-0192, Okayama, Japan;Department of Clinical Oncology, Kawasaki Medical School, Okayama, Japan;Department of Molecular and Developmental Biology, Kawasaki Medical School, Okayama, Japan;Department of Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan;

关键词: Migration;    Chemotaxis;    Lipid mediator;    Inhibitor;    TAXIScan;    Metastasis;   

DOI  :  10.1186/s12885-017-3218-4

 received in 2016-03-14, accepted in 2017-03-22,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundMigration 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.MethodsPancreatic 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.ResultsScaffold 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 (IC50 for 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. LPA1 mRNA was highest among the 6 receptors, and LPA1, LPA2 and LPA3 proteins 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.ConclusionsWe 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.

【 授权许可】

CC BY   
© The Author(s). 2017