Journal: Research

Article Title: IFI44 Promotes Clear Cell Renal Cell Carcinoma Progression via PRDX1 and Predicts Poor Prognosis

doi: 10.34133/research.1102

Figure Lengend Snippet: IFI44 knockdown inhibits malignant phenotypes in RCC cells. (A) IFI44 levels in RCC cell lines assessed by Western blot with β-actin as the control. (B) IFI44 protein levels assessed by Western blot in Caki-2 and 786-O cells following IFI44 knockdown (KD) or negative control (NC). (C and D) Densitometric quantification of IFI44 levels in Caki-2 (C) and 786-O (D) cells normalized to β-actin. (E) CCK-8 assay showing Caki-2 and 786-O cell proliferation at 0, 24, 48, and 72 h. (F) Flow cytometric analysis and quantification of Caki-2 and 786-O cell apoptosis following IFI44 KD. (G) Representative wound healing images and quantification of migration rates of Caki-2 and 786-O cells after KD or NC treatment. (H) Representative images of cell migration and invasion in Caki-2 and 786-O after KD or NC treatment, with corresponding quantification of migrated/invaded cells shown below.

Article Snippet: The human RCC cell lines 786-O (TCHu186), Caki-1 (TCHu135), Caki-2 (TCHu251), and ACHN (TCHu199) were sourced from the American Type Culture Collection in Shanghai, China.

Techniques: Knockdown, Western Blot, Control, Negative Control, CCK-8 Assay, Migration

Journal: Theranostics

Article Title: Specific in vivo detection of V2R-positive metastatic ccRCC using a toxin-based PET radioligand

doi: 10.7150/thno.126311

Figure Lengend Snippet: In vivo visualization of V2R + tumors in mccRCC representative mice models. A: Representative composite images taken at 4 h after the i.v. injection of 20 nmol/kg for the [ 18 F]F-MQ232 in CHO-304 xenografted NMRI-Foxn1 nu/nu mice (6.8 ± 1.1 MBq, 274 ± 43 MBq/kg), CHO-3013 xenografted NMRI-Foxn1 nu/nu mice (3.8 ± 1.2 MBq, 150 ± 47 MBq/kg), Caki-1 xenografted NMRI-Foxn1 nu/nu mice (3.8 ± 1.2 MBq, 152 ± 47 MBq/kg) and Renca grafted BALB/cJ mice (2.5 ± 0.9 MBq, 99 ± 37 MBq/kg). Images shown are the result of a fusion between the Computed Tomography (CT) image (grey scale) and the PET image (false colors). B: Summary table of the main PET imaging indicators obtained after 4 hours of the i.v. injection of 20 nmol/kg for the [ 18 F]F-MQ232 in CHO-304 xenografted NMRI-Foxn1 nu/nu mice (6.8 ± 1.1 MBq, 274 ± 43 MBq/kg), CHO-3013 xenografted NMRI-Foxn1 nu/nu mice (3.8 ± 1.2 MBq, 150 ± 47 MBq/kg), Caki-1 xenografted NMRI-Foxn1 nu/nu mice (3.8 ± 1.2 MBq, 152 ± 47 MBq/kg) and Renca grafted BALB/cJ mice (2.5 ± 0.9 MBq, 99 ± 37 MBq/kg). C: Flow cytometry results obtained on freshly dissociated Caki-1 and Renca cells stained using 100 nM Cy5-MQ232 (total signal, red curve) or 100 nM Cy5-MQ232 in presence of 30 µM of MQ232 (non-specific signal, blue curve). Light gathered at 680 nm, 30,000 events per acquisition, two acquisitions per independent experiment, n = 3). D: Quantification results of AVPR2 -directed RT-qPCR performed on Caki-1 and Renca cells and tumors resulting from their implantation. Results are shown as fold changes between the expression in the investigated samples and the expression in the positive reference, healthy human total kidney. E: Quantification results of AVPR2 -directed RT-qPCR performed on 8 ccRCC and mccRCC biopsies (P1 to P8). Three samples came from low grade primary tumors (grade ≤ 2) (P1 to P3, in green), two from high grade primary tumors (grade 3 or 4) (P4 and P5, in blue), and three from distant metastases (P6 to P8, in purple). Results are shown as fold changes between the expression in the investigated samples and the expression in the positive reference, healthy human total kidney.

Article Snippet: Caki-1 cells (human ccRCC, HTB-46, ATCC, USA) were cultured in McCoy's 5A supplemented with 10% FBS +1% penicillin/streptomycin.

Techniques: In Vivo, Injection, Computed Tomography, Imaging, Flow Cytometry, Staining, Quantitative RT-PCR, Expressing

Journal: Journal of Translational Medicine

Article Title: CCL5 promotes the epithelial-mesenchymal transition of circulating tumor cells in renal cancer

doi: 10.1186/s12967-024-05297-2

Figure Lengend Snippet: The effect of CCL5 on the phenotype and molecular aspects of CAKI-2 cells. A Transwell assays evaluating the effect of CCL5 on the migration and invasion capabilities of CAKI-2 cells. B Above: Quantitative analysis of migrated cell numbers in CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. Below: Quantitative analysis of invasive cell numbers in CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. C Colony formation assays assessing the impact of CCL5 on the cloning efficiency of CAKI-2 cells. D Quantitative analysis of colony numbers in CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. E Cell proliferation assays for CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. F Flow cytometry analysis of the effect of CCL5 on the cell cycle of CAKI-2 cells. G Quantitative analysis of cell cycle phase distribution changes in CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. H Flow cytometry analysis of CCL5's effect on apoptosis in CAKI-2 cells. I Quantitative analysis of apoptosis ratios in CAKI-2-CCL5-Con and CAKI-2-CCL5-shRNA groups. J RT-qPCR analysis of the effect of CCL5 on EMT-related markers in CAKI-2 cells. K Western blot analysis of the effect of CCL5 on the expression of EMT-related markers and smad2/3 molecules in CAKI-2 cells. L Immunofluorescence assay to detect the impact of CCL5 on E-cadherin expression in CAKI-2 cells. Experiments involving quantitative analysis were repeated at least three times, with statistical analysis conducted using the t-test

Article Snippet: The renal cancer cell lines, 786-O and CAKI-2, were sourced from Procell Biotechnology Co (China).

Techniques: Migration, shRNA, Clone Assay, Flow Cytometry, Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence