title: Niclosamide suppresses migration of hepatocellular carcinoma cells and downregulates matrix metalloproteinase-9 expression.
abstract: Metastasis negatively affects the prognosis of hepatocellular carcinoma (HCC). In the present study, niclosamide, which is known to suppress the proliferation of HCC cells, was investigated for possible suppressant effects on the migration of HCC cells. HLF and PLC/PRF/5 HCC cells were cultured in the presence of niclosamide. Cell proliferation was analyzed using the MTS assay. Cell migration was measured by performing a scratch assay. Expression levels of cyclin D1 and matrix metalloproteinase 9 (MMP9) were analyzed by performing revers transcription-quantitative polymerase chain reaction. Compared with the control treatment, treatment with 10 µm niclosamide suppressed the proliferation of the HLF and PRL/PRF/5 cells to 49.9±3.7 and 17.9±11.5% (P<0.05), respectively. Furthermore, compared with the control treatment, treatment with 1.0 µM niclosamide downregulated the expression of cyclin D1 to 52.4±4.4 and 23.9±5.4% (P<0.05) in the HLF and PRL/PRF/5 cells, respectively. In the scratch assay, treatment of the HLF cells with niclosamide (1.0 µm) decreased the distance of the scratched line from the growing edge to 4.6±1.0 mm compared with the 9.2±1.4 mm observed with the control treatment (P<0.05). Similarly, treatment of the PRL/PRF/5 cells with niclosamide (1.0 µm) also decreased the distance of the scratched line from the growing edge to 3.0±0.8 mm compared with the 5.5±0.9 mm observed with the control treatment (P<0.05). Further, MMP9 expression levels in the HLF cells treated with 1.0 µm niclosamide decreased to 22.4±1.76% (P<0.05) compared with those in the untreated control HLF cells. Similarly, expression level of MMP9 in the PRL/PRF/5 cells treated with 1.0 µm niclosamide deceased to 18.7±10.7% (P<0.05) compared with those in the untreated control PRL/PRF/5 cells. Overall, niclosamide downregulated the expression of MMP9 in and suppressed the migration of HCC cells.
authors.affiliation: Department of Gastroenterology, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Tomizawa
authors.affiliation: Department of Radiology, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Fuminobu
authors.initials: F
authors.lastname: Shinozaki
authors.affiliation: Department of Neurology, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Motoyoshi
authors.affiliation: Department of Rheumatology, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Sugiyama
authors.affiliation: Department of Pediatrics, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Shigenori
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Neurosurgery, National Hospital Organization Shimoshizu Hospital, Yotsukaido, Chiba 284-0003, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Ishige
keywords: MTS assay; quantitative polymerase chain reaction; scratch assay