Journal: Cancer research

Article Title: MDMX recruits UbcH5c to facilitate MDM2 E3 ligase activity and subsequent p53 degradation in vivo

doi: 10.1158/0008-5472.CAN-20-0790

Figure Lengend Snippet: (A) MEFs of indicated genotypes were treated with 4-OHT for 2 hours, cell lysates were harvested and immunoprecipitated (IP) for MDM2 or MDMX, and blotted for indicated proteins. Input represents 5% of total cell lysate utilized for IP. (B) MEFs of indicated genotypes were treated as in A and immunoprecipitated for MDM2 and MDMX and blotted for indicated proteins. (C) SJSA and MCF-7 cells were immunoprecipitated (IP) for MDM2 or MDMX, and immunoblotted with indicated antibodies. Input represents 5% of total cell lysate utilized for IP. (D) MEFs of indicated genotypes were treated with siRNA against Mdmx (MDMX) or non-specific sequences (NS) for 24 hour followed by treatment with 4-OHT for 2 hours. Cell lysates were harvested and immunoprecipitated for MDM2 and blotted for indicated proteins. (E) MDMX expressed in bacterial was analyzed by SDS-PAGE and Commassie Blue staining. (F) MDMX-UbcH5c complexes were resolved by SDS-PAGE and blotted for MDMX, GST, and UbcH5c.

Article Snippet: Cell lines The human osteosarcoma SJSA, osteosarcoma U2OS, and breast cancer MCF7 cells were obtained from the ATCC and came with comprehensive authentication and quality controls.

Techniques: Immunoprecipitation, SDS Page, Staining

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Upregulation of microRNA-23b-3p induced by farnesoid X receptor regulates the proliferation and apoptosis of osteosarcoma cells

doi: 10.1186/s13018-019-1404-6

Figure Lengend Snippet: Correlation of farnesoid X receptor (FXR) and microRNA-23b-3p expression in osteosarcoma (OS) cells and miR-23b-3p specifically targets cyclin G1 (CCNG1). As FXR might regulate the expression of miR-23b-3p in OS cells, we measured the expressions of FXR and miR-23b-3p in normal osteoblasts (hFOB1.19) and five osteosarcoma cell lines (MG-63, HOS, U2OS, SAOS2, SJSA1). a The relative expression levels of FXR were much downregulated in OS cell lines, especially in MG-63 cells. b The expressions of miR-23b-3p were obviously decreased, especially in MG-63 cells compared with hFOB1.19 cells. c Scatter plots showed the correlation of FXR and miR-23b-3p expression in normal osteoblasts and five osteosarcoma cell lines (Pearson’s coefficient test R 2 = 1.00, P = 0.0028). d The changes of miR-23b-3p levels were measured under different concentrations of the FXR agonist, GE4064 (0, 0.5, and 5 μM). e TargetScan predicted that the fragment of CCNG1-3′-UTR contained a binding site of miR-23b-3p. f The correlation between miR-23b-3p and CCNG1 was verified by luciferase reporter assay. Each value represents mean ± SEM ( n = 3). GAPDH and U6 served as internal controls for cellular genes and miR-23b-3p, respectively. ** p < 0.01 vs. hFOB1.19 cells or blank group; ## p < 0.01 vs. 0.5 μM GW4064 group

Article Snippet: Normal human osteoblasts (hFOB1.19) and osteosarcoma cell lines (MG-63, HOS, U2OS, SAOS2, and SJSA1) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: Upregulation of microRNA-23b-3p induced by farnesoid X receptor regulates the proliferation and apoptosis of osteosarcoma cells

doi: 10.1186/s13018-019-1404-6

Figure Lengend Snippet: GW4064 treatment inhibited cell proliferation and induced apoptosis and cell cycle arrest in MG-63 cells. We further study the effects of farnesoid X receptor (FXR) regulation on the development of MG-63 cells and the role of microRNA-23b-3p during the progress. a , b In order to determine the effects of FXR regulation on osteosarcoma (OS) cell proliferation, clone formation assay of MG-63 cells were performed under the single and combined effects of miR-23b-3p inhibition and GW4064 treatment. c , d The changes of apoptosis rate were analyzed by flow cytometry. e , f The cell cycle distribution of MG-63 cells was determined by flow cytometry. g , h The protein levels of several apoptotic genes (B-cell lymphoma-2 (Bcl-2), Bax, and Cleaved Caspase-3 (C caspase-3)) were analyzed by Western blot to further assess the effects of FXR regulation on OS cell apoptosis. Each value represents mean ± SEM ( n = 3). GAPDH served as an internal control. * p < 0.05, ** p < 0.01 vs. IC group; ## p < 0.01 vs. IC + GW4064 group; ^^ p < 0.01 vs. I group

Article Snippet: Normal human osteoblasts (hFOB1.19) and osteosarcoma cell lines (MG-63, HOS, U2OS, SAOS2, and SJSA1) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Tube Formation Assay, Inhibition, Flow Cytometry, Western Blot, Control

Journal: Cancer Management and Research

Article Title: A Novel Predictive Model Associated with Osteosarcoma Metastasis

doi: 10.2147/CMAR.S332387

Figure Lengend Snippet: The qPCR result of seven lncRNAs in five OS highly metastasis and weakly metastasis cell lines. ( A ) AL512422.1. ( B ) AL008718.3. ( C ) AC006033.2. ( D ) AL357507.1. ( E ) AL360182.2. ( F ) C5orf66-AS1. ( G ) CEBPA-DT. *<0.05, **<0.01, ***<0.001, ****<0.0001.

Article Snippet: Highly metastasis human OS cell lines (MNNG) and weakly metastasis human OS cell lines (U2S2, SAOS-2, SJSA-1, and HOS) were purchased from iCell (Shanghai, China).

Techniques: