control osrc2 cells  (Thermo Fisher)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Expressing, DNA Methylation Assay, Sequencing, Comparison, Methylation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 overexpression attenuated cell proliferation and induced cell apoptosis in vitro and in vivo. a and b EdU assay and c CCK-8 assay determined cell proliferation of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the ZNF582-AS1-overexpressed and control groups were measured and compared. (k and l) The cell proliferation and apoptosis of tumors were examined

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, In Vitro, In Vivo, EdU Assay, CCK-8 Assay, Control, TUNEL Assay, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 overexpression inhibited cell migratory and invasive ability in vitro and in vivo. a and b Wound healing assay determined the migratory distances of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the ZNF582-AS1-overexpressed group were remarkably lower than those in the control group. i There was no significant difference between the mice weight of ZNF582-AS1-overexpressed and control group. j and k The number and size of pulmonary metastases in the ZNF582-AS1-overexpressed group were significantly reduced compared with those in the control group. l and m ZNF582-AS1 overexpression increased E-cadherin expression and decreased N-cadherin expression in pulmonary metastases

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, In Vitro, In Vivo, Wound Healing Assay, Control, Migration, Luciferase, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 overexpression decreased rRNA adenine N(6)-methyltransferase A8K0B9 expression. a The overexpression efficiency of ZNF582-AS1 in OSRC2 cells. b iTRAQ results showed that 69 proteins were downregulated and 75 proteins were upregulated in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. c The expression of these 144 differently expressed proteins in ZNF582-AS1-overexpressed OSRC2 cells and control cells. d Biological Process GO term enrichment analysis of the complete 144 statistically significant proteins. e and f Validation of the presence of A8K0B9 protein. g A8K0B9 expression was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. h Changes of A8K0B9 protein level in OSRC2 cells after transient transfection of different doses of ZNF582-AS1 overexpression plasmid. i Estimation of the binding propensity of A8K0B9-ZNF582-AS1 pair. j RNA pull-down assay indicated that A8K0B9 protein bound specifically to ZNF582-AS1

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, Expressing, Multiplex sample analysis, Control, Biomarker Discovery, Transfection, Plasmid Preparation, Binding Assay, Pull Down Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 regulated the N(6)-methyladenosine modification of MT-RNR1 by modulating A8K0B9. a Negative and positive controls of rRNA MeRIP-seq. b The analysis results of rRNA MeRIP-seq. c The expression of MT-RNR1, MT-RNR2 and RNA28SN5 in ZNF582-AS1-overexpressed and control OSRC2 cells. d and e MT-RNR1 expression was negatively associated with ZNF582-AS1 expression ( n = 530), and lower MT-RNR1 expression was related to longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-RNR1 expression, patients were divided into two groups for survival analysis. f The expression of MT-RNR1 in ccRCC cell lines. g Analysis of m6A motif DRACH (D = A, G or U; R = A or G; H = A, U or C) in MT-RNR1 sequence. h MeRIP-qPCR determined the methylation level of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells and control cells. i RIP-RT-qPCR showed that A8K0B9 protein had a certain binding ability with MT-RNR1 in OSRC2 cells

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Modification, Expressing, Control, Sequencing, Methylation, Quantitative RT-PCR, Binding Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: ZNF582-AS1 overexpression decreased MT-CO2 expression by regulating MT-RNR1. a and b MT-CO2 expression was significantly positively correlated with MT-RNR1 expression (n = 530), and lower MT-CO2 expression was associated with longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-CO2 expression, patients were divided into two groups for survival analysis. c Based on the iTRAQ results, the expression level of MT-CO2 (A0A0P0C1B5) protein was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. d and e MT-CO2, Bcl-2 and N-cadherin protein expression was decreased and Cleaved Caspase 3 and E-cadherin protein expression was increased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. f and g The ROS level in ZNF582-AS1-overexpressed OSRC2 cells was increased. h The overexpression efficiency of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells. i and j MT-CO2, Bcl-2 and N-cadherin protein expression was increased and Cleaved Caspase 3 and E-cadherin protein expression was decreased in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. k and l The ROS level in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells was decreased

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, Expressing, Multiplex sample analysis, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: MT-RNR1 overexpression reversed inhibited cell proliferation and increased cell apoptosis caused by ZNF582-AS1 overexpression. a and b EdU assay and c CCK-8 assay determined cell proliferation of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control groups were measured and compared. k and l The cell proliferation and apoptosis of tumor were examined

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, EdU Assay, CCK-8 Assay, Control, TUNEL Assay, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

doi: 10.1186/s13046-021-01889-8

Figure Lengend Snippet: MT-RNR1 overexpression reversed inhibited cell migratory and invasive ability caused by ZNF582-AS1 overexpression. a and b Wound healing assay determined the migratory distances of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were remarkably higher than those in the ZNF582-AS1-overexpressed group. i There was no significant difference between the mouse weight of the two treatment group. j and k The number and size of pulmonary metastases in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were significantly increased compared with those in the ZNF582-AS1-overexpressed group. l and m MT-RNR1 overexpression decreased E-cadherin expression and increased N-cadherin expression in pulmonary metastases

Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

Techniques: Over Expression, Wound Healing Assay, Control, Migration, Luciferase, Expressing