Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Expression of REGγ in RCC tissues (T) and normal kidney tissues (N) as detected by IHC ( a ) and WB ( b ). c Expression of REGγ in four RCC cell lines (A498, 786-O, ACHN, and caki-1) and the normal renal tubular epithelial cell line (HK-2) as detected by western blot. d Comparison of REGγ expression in different Fuhrman grades I–IV of RCC tissue samples via IHC staining (left panel: magnification ×200, scale bar = 50 μm; right panel: magnification ×400, scale bar = 20 μm). e REGγ (PSME3) expression (median of expression intensity) in different pathological types and grades of RCC derived from Oncomine database ( https://www.oncomine.org/ ). f Kaplan–Meier analysis of the correlation between REGγ expression and the survival time in RCC patients. Cases were classified into lower expression group and higher expression group as described in methods. g Prognosis of RCC (KIRC) patients with high or low expression of REGγ (PSME3) derived from OncoLnc database ( https://www.oncolnc.org/ )

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, Western Blot, Immunohistochemistry, Derivative Assay

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a PSME3 upregulation is closely correlated with renal cell carcinoma revealed by KEGG pathway analysis. b GSEA indicated that cell proliferation and anti-apoptosis were positively correlated with elevated PSME3 expression in RCC from database GSE89563 (GO_0008284 and GO_0006915). NES, normalized enrichment score. c Stable knockdown of REGγ (shREGγ) in RCC cell lines (ACHN and A498) confirmed by WB. d , e Effect of shREGγ on RCC cells growth as determined by MTT assay. f Representative images of colony formation of RCC cells after transfection of shREGγ versus shNC. g , h Representative images of EdU incorporation assay after transfection of shREGγ versus shNC. i , j Representative flow cytometry plots of cell cycle distribution from ACHN and A498 cells transfected with shREGγ and shNC. k , l Apoptosis rate from ACHN and A498 cells after transfected with shREGγ and shNC as detected by flow cytometry. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, MTT Assay, Transfection, Flow Cytometry

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Representative images and the relative quantification of wound-healing assay in RCC cells transfected with shREGγ and shNC. c , d Representative images and the relative quantification of transwell invasion assay in ACHN and A498 cells transfected with shREGγ and shNC. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Wound Healing Assay, Transfection, Transwell Invasion Assay

Journal: Molecular Oncology

Article Title: Obesity alters the fitness of peritumoral adipose tissue, exacerbating tumor invasiveness in renal cancer through the induction of ADAM12 and CYP1B1

doi: 10.1002/1878-0261.13782

Figure Lengend Snippet: Adipose peritumoral tissue affects the expression of inflammatory genes by renal cell carcinoma (RCC) cells. (A) Relative mRNA expression of the different tumor suppressor genes in the healthy and tumor kidney tissues of RCC patients ( n = 10), including VHL , PBRM1 , SETD2 , and BAP1 , was measured by real‐time PCR. Cyclophilin‐A was used as an internal control. Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by Student's t ‐test. (B) Schematic figure illustrating the protocol for differentiation of adipocytes from peritumoral adipose tissue (AT) of lean, overweight, and obese RCC patients. (C) Relative mRNA expression of the leptin and adiponectin of the differentiated adipocytes among the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. (D) Schematic figure of the conditioned medium (CM) experiment, showing RCC cells treated with conditioned media (CMs) of lean, overweight, and obese differentiated adipocytes. (E) Relative mRNA expression of the IL6 , CXCR4 , SDF1 , and BAFFR of the A498 cells treated with CMs collected from differentiated adipocytes derived from the lean, overweight, and obese groups ( n = 6). Data are shown as mean ± SD from at least 3 separate experiments. P ‐values were determined by one‐way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The human renal cancer cell line (A498) was grown in DMEM (Microgem); supplemented with 10% FBS, 1% Pen/Strep, and 1% L‐Glutamine, at 37 °C in 5% CO 2 .

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Derivative Assay