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[Effects of phosphatase of regenerating liver cell-3 gene silence by RNA interference on the expression of matrix metalloproteinases-2,-9 in human colon cancer cells].

Yu Fan, Youli Zhang, Shu Sen Zheng

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Abstract

OBJECTIVE: To explore the effects and associated mechanism of phosphatase of regenerating liver cell-3 (PRL-3) on the invasion of human colon cancer cell. METHODS: After colon cancer cell line HCT116 was transfected with PRL-3 small interfering RNA (siRNA), the mRNA and protein expression of PRL-3 and matrix metalloproteinase 2 (MMP-2) and MMP-9 were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined using clone formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Then the transfected HCT116 cells were implanted into nude mice and the tumor growth was observed. RESULTS: PRL-3 siRNA could inhibit anchorage-independent growth of HCT116 cells in a dose-dependent manner in vitro. The mRNA and protein expression of MMP-2 and MMP-9 were down-regulated by PRL-3 siRNA. HCT116 cells invaded striated muscle and vessels in control nude mice but such phenomena were not found in transfected HCT116-implanted nude mice in vivo. CONCLUSION: PRL-3 siRNA inhibit the invasion of colon cancer cells possibly through the down-regulation of MMP-2 and MMP-9.

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OBJECTIVE: To explore the effects and associated mechanism of phosphatase of regenerating liver cell-3 (PRL-3) on the invasion of human colon cancer cell. METHODS: After colon cancer cell line HCT116 was transfected with PRL-3 small interfering RNA (siRNA), the mRNA and protein expression of PRL-3 and matrix metalloproteinase 2 (MMP-2) and MMP-9 were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined using clone formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Then the transfected HCT116 cells were implanted into nude mice and the tumor growth was observed. RESULTS: PRL-3 siRNA could inhibit anchorage-independent growth of HCT116 cells in a dose-dependent manner in vitro. The mRNA and protein expression of MMP-2 and MMP-9 were down-regulated by PRL-3 siRNA. HCT116 cells invaded striated muscle and vessels in control nude mice but such phenomena were not found in transfected HCT116-implanted nude mice in vivo. CONCLUSION: PRL-3 siRNA inhibit the invasion of colon cancer cells possibly through the down-regulation of MMP-2 and MMP-9.

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Available abstract

OBJECTIVE: To explore the effects and associated mechanism of phosphatase of regenerating liver cell-3 (PRL-3) on the invasion of human colon cancer cell. METHODS: After colon cancer cell line HCT116 was transfected with PRL-3 small interfering RNA (siRNA), the mRNA and protein expression of PRL-3 and matrix metalloproteinase 2 (MMP-2) and MMP-9 were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined using clone formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Then the transfected HCT116 cells were implanted into nude mice and the tumor growth was observed. RESULTS: PRL-3 siRNA could inhibit anchorage-independent growth of HCT116 cells in a dose-dependent manner in vitro. The mRNA and protein expression of MMP-2 and MMP-9 were down-regulated by PRL-3 siRNA. HCT116 cells invaded striated muscle and vessels in control nude mice but such phenomena were not found in transfected HCT116-implanted nude mice in vivo. CONCLUSION: PRL-3 siRNA inhibit the invasion of colon cancer cells possibly through the down-regulation of MMP-2 and MMP-9.

Key concepts: Transfection, Matrix metalloproteinase, Small interfering RNA, Molecular biology, Cell culture, RNA interference, Messenger RNA, Medicine

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[Effects of phosphatase of regenerating liver cell-3 gene silence by RNA interference on the expression of matrix metalloproteinases-2,-9 in human colon cancer cells]. — Research Paper | ScholarLens