suppressor serum  (Vector Laboratories)

Journal: Oncology Letters

Article Title: miR-142-5p suppresses proliferation and promotes apoptosis of human osteosarcoma cell line, HOS, by targeting PLA2G16 through the ERK1/2 signaling pathway

doi: 10.3892/ol.2018.9712

Figure Lengend Snippet: The expression level of PLA2G16 protein and miR-142-5p in human OS tissues and adjacent healthy tissues, in addition to human hFOB1.19 cells and HOS cells. (A) miR-142-5p expression level in OS tissues compared with adjacent healthy tissues. (B) miR-142-5p expression level of human hFOB1.19 cells compared with HOS cells. (C) PLA2G16 protein expression level in OS tissues compared with adjacent healthy tissues. (D) PLA2G16 protein expression level of human hFOB1.19 cells compared with HOS cells. *P<0.05 compared with normal tissues or hFOB1.19 cells. OS, osteosarcoma; miR, microRNA; PLA2G16, group XVI phospholipase A2.

Article Snippet: Abbreviations OS Osteosarcoma miRNAs microRNAs 3′UTR 3′-untranslated region DMEM Dulbecco's modified Eagle's medium PLA2G16 Group XVI phospholipase A2 HRASLS3 Ha-RAS like suppressor 3 AdPLA2 adipose specific PLA2 ATCC American Type Culture Collection MTT modified tetrazolium salt 3-(4,5-dimethyl-2-thiazolyl)-2,5-dipheny-2 H-tetrazolium-bromide DMSO dimethyl sulfoxide BrdU 5-Bromo-2′-deoxyuridine PVDF polyvinylidene fluoride membranes RT-qPCR reverse transcription-quantitative polymerase chain reaction SD standard deviation LPA lysophosphatidic acid FBS fetal bovine serum FFA free fatty acid ELISA enzyme-linked immunosorbent assay MAPK/ERK mitogen-activated protein kinases/extracellular signal-regulated kinase

Techniques: Expressing

Journal: Oncology Letters

Article Title: miR-142-5p suppresses proliferation and promotes apoptosis of human osteosarcoma cell line, HOS, by targeting PLA2G16 through the ERK1/2 signaling pathway

doi: 10.3892/ol.2018.9712

Figure Lengend Snippet: PLA2G16 is a direct target gene of miR-142-5p. (A) A microRNA database search predicted an miR-142-5p binding site on the PLA2G16 3′-UTR. (B) miR-142-5p overexpression in 293T cells reduced the luciferase activity of cells transfected with pMIR-PLA2G16-wt 3′-UTR, whereas pMIR-PLA2G16-mut 3′-UTR blocked this effect. (C) Following transfection with miR-142-5p mimics or miR-NC, miR-142-5p transfection induced an elevation in the expression level of mature miR-142-5p in HOS cells. (D) The expression level of PLA2G16 protein in HOS cells was detected and measured 72 h following transfection with miR-142-5p and miR-NC mimics. *P<0.05 compared with control. miR, microRNA; PLA2G16, group XVI phospholipase A2; NC, negative control; UTR, untranslated region; wt, wild type; mut, mutated.

Article Snippet: Abbreviations OS Osteosarcoma miRNAs microRNAs 3′UTR 3′-untranslated region DMEM Dulbecco's modified Eagle's medium PLA2G16 Group XVI phospholipase A2 HRASLS3 Ha-RAS like suppressor 3 AdPLA2 adipose specific PLA2 ATCC American Type Culture Collection MTT modified tetrazolium salt 3-(4,5-dimethyl-2-thiazolyl)-2,5-dipheny-2 H-tetrazolium-bromide DMSO dimethyl sulfoxide BrdU 5-Bromo-2′-deoxyuridine PVDF polyvinylidene fluoride membranes RT-qPCR reverse transcription-quantitative polymerase chain reaction SD standard deviation LPA lysophosphatidic acid FBS fetal bovine serum FFA free fatty acid ELISA enzyme-linked immunosorbent assay MAPK/ERK mitogen-activated protein kinases/extracellular signal-regulated kinase

Techniques: Binding Assay, Over Expression, Luciferase, Activity Assay, Transfection, Expressing, Control, Negative Control

Journal: Oncology Letters

Article Title: miR-142-5p suppresses proliferation and promotes apoptosis of human osteosarcoma cell line, HOS, by targeting PLA2G16 through the ERK1/2 signaling pathway

doi: 10.3892/ol.2018.9712

Figure Lengend Snippet: Effects of miR-142-5p and PLA2G16 on p-ERK1/2 protein expression, proliferation and apoptosis of HOS cells. HOS cells were transfected with miR-142-5p, miR-NC mimics and pcDNA3.1-PLA2G16 plasmids. (A) The expression level of p-Raf-1, Raf-1, p-MEK, MEK, p-ERK1/2, ERK1/2 and β-actin proteins. miR-142-5p transfection decreased the expression levels of p-Raf-1, p-MEK, and p-ERK1/2 proteins in HOS cells. (B) The expression levels of PLA2G16, p-ERK1/2, ERK1/2 and β-actin proteins were detected and measured. The PLA2G16 expression level was enhanced in HOS cells following transfection with pCDNA3.1-PLA2G16 and PLA2G16 overexpression rescued the reduced expression level of p-ERK1/2 protein associated with miR-142-5p transfection in HOS cells. (C) Apoptotic rate of HOS cells following transfection with miR-142-5p mimics either with or without pCDNA3.1-PLA2G16 at 24, 48 and 72 h. (D) CASP3 activity assays were conducted in HOS cells following transfection with miR-142-5p mimics either with or without pCDNA3.1-PLA2G16. *P<0.05 compared with control. #P<0.05 compared with miR-142-5p mimics. OD, optical density; p, phosphorylated; Raf-1, proto-oncogene, serine/threonine kinase 1, CASP3, Caspase-3; ERK1/2, extracellular signal-regulated kinase 1/2; MEK, mitogen-activated protein kinase kinase; NC, negative control; PLA2G16, group XVI phospholipase A2.

Article Snippet: Abbreviations OS Osteosarcoma miRNAs microRNAs 3′UTR 3′-untranslated region DMEM Dulbecco's modified Eagle's medium PLA2G16 Group XVI phospholipase A2 HRASLS3 Ha-RAS like suppressor 3 AdPLA2 adipose specific PLA2 ATCC American Type Culture Collection MTT modified tetrazolium salt 3-(4,5-dimethyl-2-thiazolyl)-2,5-dipheny-2 H-tetrazolium-bromide DMSO dimethyl sulfoxide BrdU 5-Bromo-2′-deoxyuridine PVDF polyvinylidene fluoride membranes RT-qPCR reverse transcription-quantitative polymerase chain reaction SD standard deviation LPA lysophosphatidic acid FBS fetal bovine serum FFA free fatty acid ELISA enzyme-linked immunosorbent assay MAPK/ERK mitogen-activated protein kinases/extracellular signal-regulated kinase

Techniques: Expressing, Transfection, Over Expression, Activity Assay, Control, Negative Control