MicroRNA-222 contributed to cell proliferation, invasion and migration via regulating YWHAG in osteosarcoma.
Y‐W Chu, C-R Wang, Weng Fb, Yan Zj, Changsong Wang
Abstract
Y‐W Chu, C-R Wang, Weng Fb, Yan Zj, Changsong Wang
Abstract
OBJECTIVE: The aim of this study was to investigate the role of microRNA-222 (miR-222) in osteosarcoma (OS), and to further explore the potential molecular mechanism. PATIENTS AND METHODS: We measured the level of miR-222 in OS tissues and cell lines using quantitative Real-time polymerase chain reaction. Synthesized miR-222 mimics or inhibitors were obtained to up-regulate or down-regulate the expression of miR-222 in U2OS or Saos2 cells. Cell counting kit-8 (CCK8) and colony formation assay were employed to detect the ability of cell proliferation, and transwell assay was used to confirm the ability of cell invasion. Furthermore, luciferase assay and Western blot were applied to verify the target of miR-222 in OS. RESULTS: The level of miR-222 in OS tumor tissue samples was significantly lower than that in normal group. Over-expression of miR-222 decreased cell proliferation and invasion in U2OS cells while knockdown of miR-222 promoted cell growth and metastasis in Saos2 cells. Furthermore, YWHAG was found to be a candidate target of miR-222 using several databases. Elevated level of miR-222 inhibited YWHAG expression while reduced miR-222 promoted YWHAG expression. Also, up-regulation of YWHAG restored the inhibiting effect of miR-222 mimics. CONCLUSIONS: We identified for the first time that the expression level of miR-222 was reduced in OS tissues as well as in OS cell lines. miR-222 could inhibit cell proliferation and invasion via down-regulating YWHAG. These data could provide a potential target for the biological treatment of OS.
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OBJECTIVE: The aim of this study was to investigate the role of microRNA-222 (miR-222) in osteosarcoma (OS), and to further explore the potential molecular mechanism. PATIENTS AND METHODS: We measured the level of miR-222 in OS tissues and cell lines using quantitative Real-time polymerase chain reaction. Synthesized miR-222 mimics or inhibitors were obtained to up-regulate or down-regulate the expression of miR-222 in U2OS or Saos2 cells. Cell counting kit-8 (CCK8) and colony formation assay were employed to detect the ability of cell proliferation, and transwell assay was used to confirm the ability of cell invasion. Furthermore, luciferase assay and Western blot were applied to verify the target of miR-222 in OS. RESULTS: The level of miR-222 in OS tumor tissue samples was significantly lower than that in normal group. Over-expression of miR-222 decreased cell proliferation and invasion in U2OS cells while knockdown of miR-222 promoted cell growth and metastasis in Saos2 cells. Furthermore, YWHAG was found to be a candidate target of miR-222 using several databases. Elevated level of miR-222 inhibited YWHAG expression while reduced miR-222 promoted YWHAG expression. Also, up-regulation of YWHAG restored the inhibiting effect of miR-222 mimics. CONCLUSIONS: We identified for the first time that the expression level of miR-222 was reduced in OS tissues as well as in OS cell lines. miR-222 could inhibit cell proliferation and invasion via down-regulating YWHAG. These data could provide a potential target for the biological treatment of OS.
Key concepts: Cell growth, microRNA, Gene knockdown, Western blot, Cell culture, Osteosarcoma, Cell, Luciferase