Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: Inhibitory effects of miR‐33a on cell proliferation and cell cycle in renal cell cancer (RCC) cell lines. (a) Expression of miR‐33a in Caki‐1 and 786‐O cells after transfection with miR‐33a mimics and miR‐33a inhibitor or NC. NC represents negative control of miRNA. Results were expressed as of three independent experiments, with at least three replicates in each independent experiment, ** p < 0.01 versus NC. (b) CCK‐8 assays indicated that the effects of miR‐33a on the growth of RCC cell lines. Results were expressed as of three independent experiments, with at least three replicates in each independent experiment, * p < 0.05 versus NC. (c, d) The results of flow cytometry showed that upregulation of miR‐33a significantly increased the percentages of cells in the G0/G1 phase, which showed that the upregulation of miR‐33a could suppress RCC cells proliferation. * p < 0.05 versus Control

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Expressing, Transfection, Negative Control, CCK-8 Assay, Flow Cytometry, Control

Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: miR‐33a directly targets Mouse double minute 4 (MDM4). (a) The predicted miR‐33a binding site within MDM4 3′ UTR and miR‐33a mutated version by site mutagenesis; (b) Caki‐1 cells were transfected with reporter constructs containing either wild type (WT) MDM4 , or MDM 4 3′ UTR with mutation (MUT), along with miR‐33a mimics, or negative control, respectively. Relative luciferase activity was measured. (c) qPCR analysis of MDM4 expression in renal cell cancer (RCC) Caki‐1 cells after overexpression or knockdown of miR‐33a . (d, e) Western blotting analysis of MDM4 protein expression in RCC Caki‐1 cells after overexpression or knockdown of miR‐33a . (f) Western blot analysis revealed that transfection of MDM4 siRNA into Caki‐1 cells resulted in decreased MDM4 expression compared to the cells transfected with scrambled siRNA. These effects of siRNA were attenuated by anti‐ miR‐33a inhibitor transfection. NC represents normal control, * p < 0.05, ** p < 0.01 versus NC

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Binding Assay, Mutagenesis, Transfection, Construct, Negative Control, Luciferase, Activity Assay, Expressing, Over Expression, Knockdown, Western Blot, Control

Journal: Molecular Genetics & Genomic Medicine

Article Title: miR‐33a inhibits cell growth in renal cancer by downregulation of MDM4 expression

doi: 10.1002/mgg3.833

Figure Lengend Snippet: The effect of mouse double minute 4 ( MDM4 ) on cell growth in renal cell cancer (RCC). (a, b, c) RT‐qPCR and western blot analysis of MDM4 expression in RCC cells. (d) Kaplan–Meier analysis of overall survival in 30 RCC patients with low median ( n = 15) and high median ( n = 15) expression levels of MDM4 . (e) CCK‐8 assays indicated that the effects of MDM4 on growth of RCC cell lines. Results were expressed as ± x - s of three independent experiments, with at least three replicates in each independent experiment, * p < 0.05 versus NC. (f) Western blot analysis of p53 expression after miR‐33a mimics and ‐inhibitor transfection in RCC cells

Article Snippet: Normal primary renal tubular HK‐2 cell lines and RCC cell lines (Caki‐1, ACHN and 786‐O) were purchased from China Center For Type Culture Collection (Wuhan, China).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, CCK-8 Assay, Transfection

Journal: Genetics Research

Article Title: Identification of ECI2 as Potential Prognostic Biomarkers Based on a Fatty Acid Metabolism-Related Gene Model in Clear Cell Renal Cell Carcinoma

doi: 10.1155/genr/2237539

Figure Lengend Snippet: Screening of differentially expressed genes related to fatty acid metabolism. (a) Single-cell data clustering ump plot of GSE152938 . (b) Differential expression of fatty acid metabolism gene set between tumor and adjacent tissues of ccRCC. (c) Differential expression of fatty acid metabolism gene set among different cell types. (d) Combining transcriptomics, proteomics, and single-cell genomics to screen FA-DEGs. (e) Protein interaction networks of 10 FA-DEGs using GENEMIA database. (f) Differential expression of 10 FA-DEGs in TCGA transcriptome. Data were shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Human-derived ccRCC cell lines (OS-RC-2, 769P, 786-O) and normal renal tubular epithelial cell line (HK-2) were used, all of which were purchased from ServiceBio.

Techniques: Quantitative Proteomics

Journal: Genetics Research

Article Title: Identification of ECI2 as Potential Prognostic Biomarkers Based on a Fatty Acid Metabolism-Related Gene Model in Clear Cell Renal Cell Carcinoma

doi: 10.1155/genr/2237539

Figure Lengend Snippet: Prognostic model related to fatty acid metabolism. (a) Cross-validation plot for the penalty term. (b) Plots for LASSO expression coefficients of the FA-DEGs. (c) Risk score, survival time, and survival status of ccRCC patients in TCGA. (d) The KM survival curve distribution of this risk model in ccRCC patients, where log rank is used to test between different groups. (e) Calibration curve of fatty acid metabolism–related risk model. (f) Screening key FA-DEGs using Cytoscape.

Article Snippet: Human-derived ccRCC cell lines (OS-RC-2, 769P, 786-O) and normal renal tubular epithelial cell line (HK-2) were used, all of which were purchased from ServiceBio.

Techniques: Biomarker Discovery, Expressing

Journal: Genetics Research

Article Title: Identification of ECI2 as Potential Prognostic Biomarkers Based on a Fatty Acid Metabolism-Related Gene Model in Clear Cell Renal Cell Carcinoma

doi: 10.1155/genr/2237539

Figure Lengend Snippet: Analysis of ECI2 expression levels and its correlation with clinical characteristics and prognosis. (a) ECI2 expression in paired samples in ccRCC cohort of TCGA. (b) ROC analysis of ECI2 in ccRCC cohort diagnosis. (c) Differences protein expression levels of ECI2 in CPTAC. (d) Differences phosphorylated protein expression levels of ECI2 in CPTAC. (e) Analysis of the correlation between ECI2 expression and ccRCC patients' overall survival. (f) Analysis of the correlation between ECI2 expression and ccRCC patients' disease-specific survival. (g) Analysis of the correlation between ECI2 expression and ccRCC patients' progression-free survival. (h) Correlation analysis between the expression level of ECI2 and the pathological N stage of ccRCC patients. (i) Correlation analysis between the expression level of ECI2 histologic stage of ccRCC patients. (j) Analysis of methylation levels of ECI2 at various points. (k) Correlation analysis between methylation level and mRNA expression level of ECI2 at cg11647493 site. Data were shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Human-derived ccRCC cell lines (OS-RC-2, 769P, 786-O) and normal renal tubular epithelial cell line (HK-2) were used, all of which were purchased from ServiceBio.

Techniques: Expressing, Biomarker Discovery, Methylation

Journal: Genetics Research

Article Title: Identification of ECI2 as Potential Prognostic Biomarkers Based on a Fatty Acid Metabolism-Related Gene Model in Clear Cell Renal Cell Carcinoma

doi: 10.1155/genr/2237539

Figure Lengend Snippet: Expression validation of ECI2 in clinical samples and cell lines. (a) Differences in protein expression levels of ECI2 in ccRCC tumor tissue and adjacent tissues ( n = 8). (b) Differences in mRNA expression levels of ECI2 in ccRCC tumor tissue and adjacent tissues ( n = 8). (c) Immunohistochemical staining of ccRCC tumor tissue and adjacent tissues ( n = 40). Scale bars: 200 μm. (d) Differences in protein expression levels of ECI2 in ccRCC cell lines. (e) Differences in mRNA expression levels of ECI2 in ccRCC cell lines. (f) Immunofluorescence staining confirms the subcellular localization of ECI2 (using FITC to label ECI2 and CY3 to label mitochondrial protein TOM20). Data were shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Human-derived ccRCC cell lines (OS-RC-2, 769P, 786-O) and normal renal tubular epithelial cell line (HK-2) were used, all of which were purchased from ServiceBio.

Techniques: Expressing, Biomarker Discovery, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Genetics Research

Article Title: Identification of ECI2 as Potential Prognostic Biomarkers Based on a Fatty Acid Metabolism-Related Gene Model in Clear Cell Renal Cell Carcinoma

doi: 10.1155/genr/2237539

Figure Lengend Snippet: Overexpression of ECI2 inhibits the proliferation and migration of ccRCC cells. (a) Differences in protein and mRNA expression levels of TBRG4 after lentiviral transfection into 786O. (b) The proliferation of 786O was examined by cell counts. (c) Colony formation assay was employed to evaluate the proliferation of 786O. (d) Wound healing assay measured the motor ability of 786O. (e) The transwell assay detected the migration of 786O. (f) Calculation of IC 50 differences after 48 h of treatment with different concentrations of oxaliplatin. (g) Western blot detection of protein expression differences between BCL2 and Caspase3 after 48 h of IC 50 oxaliplatin treatment. Data were shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Human-derived ccRCC cell lines (OS-RC-2, 769P, 786-O) and normal renal tubular epithelial cell line (HK-2) were used, all of which were purchased from ServiceBio.

Techniques: Over Expression, Migration, Expressing, Transfection, Colony Assay, Wound Healing Assay, Transwell Assay, Western Blot