mgmt human tagged orf  (OriGene)

Journal: Cells

Article Title: Targeting BC200/miR218-5p Signaling Axis for Overcoming Temozolomide Resistance and Suppressing Glioma Stemness

doi: 10.3390/cells9081859

Figure Lengend Snippet: Differential expression of BC200 RNA in GB cell lines. ( A ) The differential expression of BC200 RNA in GB cell lines and normal human astrocytes are shown. ( B ) The viability of GB cells was analyzed through SRB assay 48 h after TMZ (0–1000 μM) treatment. ( C ) Flow cytometry analysis of the ALDH1 + /CD133 + portion in GB cell lines and normal human astrocytes. ( D ) The level of BC200, BCRP1, MDR1, MRP1 and MGMT in GB cell lines was analyzed using RT-qPCR. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The human MGMT open reading frame (ORF) plasmid was purchased from OriGene (Cat# MGMT (RC229131, Taipei, Taiwan) and the cells transfected according to vendor’s instructions; MGMT-overexpressing U87MG cells were then allowed to grow at 37 °C in 5% CO 2 humidified atmosphere in Dulbecco’s modified Eagle’s medium (DMEM).

Techniques: Quantitative Proteomics, Sulforhodamine B Assay, Flow Cytometry, Quantitative RT-PCR

Journal: Cells

Article Title: Targeting BC200/miR218-5p Signaling Axis for Overcoming Temozolomide Resistance and Suppressing Glioma Stemness

doi: 10.3390/cells9081859

Figure Lengend Snippet: BC200 RNA promotes TMZ resistance in GB through sponge miR-218-5p. ( A ) The viability of shBC200 and OEBC200 GB cell lines was analyzed through SRB assay 48 h after TMZ (0–1000 μM) treatment. ( B ) The levels of MGMT, BCRP1, MDR1, and MRP1 following shBC200 and OEBC200 in GB cells were determined through western blot. ( C ) MicroRNA profiling analyses showed that shBC200 and OEBC200 contained high and low levels of miR-218-5p, respectively. ( D ) LncBase Predicted v.2 predicted that a high binding score of miR-218-5p with BC200. ( E ) BC200 directly interacts with multiple binding sites to hsa-miR-218-5p. *** p < 0.001.

Article Snippet: The human MGMT open reading frame (ORF) plasmid was purchased from OriGene (Cat# MGMT (RC229131, Taipei, Taiwan) and the cells transfected according to vendor’s instructions; MGMT-overexpressing U87MG cells were then allowed to grow at 37 °C in 5% CO 2 humidified atmosphere in Dulbecco’s modified Eagle’s medium (DMEM).

Techniques: Sulforhodamine B Assay, Western Blot, Binding Assay

Journal: Cells

Article Title: Targeting BC200/miR218-5p Signaling Axis for Overcoming Temozolomide Resistance and Suppressing Glioma Stemness

doi: 10.3390/cells9081859

Figure Lengend Snippet: miR-218-5p regulated cell sphere formation, colony formation, and TMZ resistance in GB cells in vitro. ( A ) The differential expression of miR-218-5p in GB cell lines and normal human astrocytes. ( B ) Pearson’s correlation curve identified a negative correlation between BC200 and miR-218-5p in GB tissues. ( C ) Sphere formation assays showed that inhibition or mock transfection of miR-218-5p regulated GB cell stemness. ( D ) Colony formation assays showed that inhibition or mock transfection of miR-218-5p regulated GB cell survival. ( E ) The viability of GB cells with inhibition or mock transfection of miR-218-5p was analyzed through SRB assay 48 h after TMZ (0–1000 μM) treatment. ( F ) CCK-8 assay showed that inhibition or mock transfection of miR-218-5p had no effect on GB cell proliferation. ( G ) The protein levels of SOX2, Oct4, BRPC1, MRP1 and MDR1 with inhibition or mock transfection of miR-218-5p. ( H ) The protein levels of MGMT, MLH1, MSH2, MSH6 and PMS2 following inhibition or mock transfection of miR-218-5p in GB cells were determined through western blot. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: The human MGMT open reading frame (ORF) plasmid was purchased from OriGene (Cat# MGMT (RC229131, Taipei, Taiwan) and the cells transfected according to vendor’s instructions; MGMT-overexpressing U87MG cells were then allowed to grow at 37 °C in 5% CO 2 humidified atmosphere in Dulbecco’s modified Eagle’s medium (DMEM).

Techniques: In Vitro, Quantitative Proteomics, Inhibition, Transfection, Sulforhodamine B Assay, CCK-8 Assay, Western Blot

Journal: Cells

Article Title: Targeting BC200/miR218-5p Signaling Axis for Overcoming Temozolomide Resistance and Suppressing Glioma Stemness

doi: 10.3390/cells9081859

Figure Lengend Snippet: The lncRNA BC200 RNA sponges miR-218-5p regulated MGMT and MMR system enhancing self-renewal and TMZ resistance of GBM cells.

Article Snippet: The human MGMT open reading frame (ORF) plasmid was purchased from OriGene (Cat# MGMT (RC229131, Taipei, Taiwan) and the cells transfected according to vendor’s instructions; MGMT-overexpressing U87MG cells were then allowed to grow at 37 °C in 5% CO 2 humidified atmosphere in Dulbecco’s modified Eagle’s medium (DMEM).

Techniques: