PDF(Pubmed)
Abstract：
UNASSIGNED: As the most common aggressive primary brain tumor, glioblastoma is inevitably a recurrent malignancy whose patients\' prognosis is poor. miR-143 and miR-145, as tumor suppressor miRNAs, are downregulated through tumorigenesis of multiple human cancers, including glioblastoma. These two miRNAs regulate numerous cellular processes, such as proliferation and migration. This research was intended to explore the simultaneous replacement effect of miR-143, and miR-145 on in vitro tumorgenicity of U87 glioblastoma cells.
UNASSIGNED: U87 cells were cultured, and transfected with miR-143-5p and miR-145-5p. Afterward, the changes in cell viability, and apoptosis induction were determined by MTT assay and Annexin V/PI staining. The accumulation of cells at the cell cycle phases was assessed using the flow cytometry. Wound healing and colony formation assays were performed to study cell migration. qRT-PCR and western blot techniques were utilized to quantify gene expression levels.
UNASSIGNED: Our results showed that miR-143-5p and 145-5p exogenous upregulation cooperatively diminished cell viability, and enhanced U-87 cell apoptosis by modulating Caspase-3/8/9, Bax, and Bcl-2 protein expression. The combination therapy increased accumulation of cells at the sub-G1 phase by modulating CDK1, Cyclin D1, and P53 protein expression. miR-143/145-5p significantly decreased cell migration, and reduced colony formation ability by the downregulation of c-Myc and CD44 gene expression. Furthermore, the results showed the combination therapy of these miRNAs could remarkably downregulate phosphorylated-AKT expression levels.
UNASSIGNED: In conclusion, miR-143 and miR-145 were indicated to show cooperative anti- cancer effects on glioblastoma cells via modulating AKT signaling as a new therapeutic approach.
UNASSIGNED: As the most common aggressive primary brain tumor, glioblastoma is inevitably a recurrent malignancy whose patients\' prognosis is poor. miR-143 and miR-145, as tumor suppressor miRNAs, are downregulated through tumorigenesis of multiple human cancers, including glioblastoma. These two miRNAs regulate numerous cellular processes, such as proliferation and migration. This research was intended to explore the simultaneous replacement effect of miR-143, and miR-145 on in vitro tumorgenicity of U87 glioblastoma cells.
UNASSIGNED: U87 cells were cultured, and transfected with miR-143-5p and miR-145-5p. Afterward, the changes in cell viability, and apoptosis induction were determined by MTT assay and Annexin V/PI staining. The accumulation of cells at the cell cycle phases was assessed using the flow cytometry. Wound healing and colony formation assays were performed to study cell migration. qRT-PCR and western blot techniques were utilized to quantify gene expression levels.
UNASSIGNED: Our results showed that miR-143-5p and 145-5p exogenous upregulation cooperatively diminished cell viability, and enhanced U-87 cell apoptosis by modulating Caspase-3/8/9, Bax, and Bcl-2 protein expression. The combination therapy increased accumulation of cells at the sub-G1 phase by modulating CDK1, Cyclin D1, and P53 protein expression. miR-143/145-5p significantly decreased cell migration, and reduced colony formation ability by the downregulation of c-Myc and CD44 gene expression. Furthermore, the results showed the combination therapy of these miRNAs could remarkably downregulate phosphorylated-AKT expression levels.
UNASSIGNED: In conclusion, miR-143 and miR-145 were indicated to show cooperative anti- cancer effects on glioblastoma cells via modulating AKT signaling as a new therapeutic approach.
摘要：
■作为最常见的侵袭性原发性脑肿瘤，胶质母细胞瘤是一种复发的恶性肿瘤，患者预后较差。miR-143和miR-145，作为肿瘤抑制miRNA，通过多种人类癌症的肿瘤发生下调，包括胶质母细胞瘤.这两种miRNA调节许多细胞过程，如扩散和迁移。本研究旨在探讨miR-143和miR-145对U87胶质母细胞瘤细胞体外致瘤性的同时替代作用。
■培养U87细胞，并用miR-143-5p和miR-145-5p转染。之后,细胞活力的变化，通过MTT法和AnnexinV/PI染色确定凋亡诱导。使用流式细胞术评估细胞周期阶段的细胞积累。进行伤口愈合和集落形成测定以研究细胞迁移。使用qRT-PCR和western印迹技术来定量基因表达水平。
■我们的结果表明，miR-143-5p和145-5p外源上调协同降低了细胞活力，并通过调节Caspase-3/8/9、Bax、和Bcl-2蛋白表达。联合疗法通过调节CDK1、细胞周期蛋白D1和P53蛋白表达来增加亚G1期细胞的积累。miR-143/145-5p显著降低细胞迁移，并通过下调c-Myc和CD44基因表达来降低集落形成能力。此外，结果显示,这些miRNA的联合治疗可以显著下调磷酸化-AKT表达水平.
■总而言之，miR-143和miR-145被表明通过调节AKT信号传导作为新的治疗方法对成胶质细胞瘤细胞显示协同抗癌作用。
■作为最常见的侵袭性原发性脑肿瘤，胶质母细胞瘤是一种复发的恶性肿瘤，患者预后较差。miR-143和miR-145，作为肿瘤抑制miRNA，通过多种人类癌症的肿瘤发生下调，包括胶质母细胞瘤.这两种miRNA调节许多细胞过程，如扩散和迁移。本研究旨在探讨miR-143和miR-145对U87胶质母细胞瘤细胞体外致瘤性的同时替代作用。
■培养U87细胞，并用miR-143-5p和miR-145-5p转染。之后,细胞活力的变化，通过MTT法和AnnexinV/PI染色确定凋亡诱导。使用流式细胞术评估细胞周期阶段的细胞积累。进行伤口愈合和集落形成测定以研究细胞迁移。使用qRT-PCR和western印迹技术来定量基因表达水平。
■我们的结果表明，miR-143-5p和145-5p外源上调协同降低了细胞活力，并通过调节Caspase-3/8/9、Bax、和Bcl-2蛋白表达。联合疗法通过调节CDK1、细胞周期蛋白D1和P53蛋白表达来增加亚G1期细胞的积累。miR-143/145-5p显著降低细胞迁移，并通过下调c-Myc和CD44基因表达来降低集落形成能力。此外，结果显示,这些miRNA的联合治疗可以显著下调磷酸化-AKT表达水平.
■总而言之，miR-143和miR-145被表明通过调节AKT信号传导作为新的治疗方法对成胶质细胞瘤细胞显示协同抗癌作用。
