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

Journal: Cell Communication and Signaling : CCS

Article Title: In renal cell carcinoma the PTEN splice variant PTEN-Δ shows similar function as the tumor suppressor PTEN itself

doi: 10.1186/s12964-018-0247-9

Figure Lengend Snippet: Expression of PTEN-Δ in RCC cell lines and translation of PTEN-Δ in RCC cells. a Relative expression values of PTEN-Δ in RCC cell lines were quantified by Real-Time PCR. Expression was normalized to expression of the house-keeping genes TBP , ATP5J and PPIA . b 786-O and A498 cells were transfected with the expression construct for the PTEN-Δ and PTEN isoform, C-terminally tagged with a V5-tag to enable detection of the PTEN-isoforms. Western blots of 786-O cell protein extracts were thus performed with an anti-V5-tag antibody. As positive control for the transfected constructs, cell free in vitro translated (IVT) protein of the respective construct was loaded. c Expression of PTEN -Δ and PTEN in transfected 786-O and A498 RCC cells were quantified by Real-Time PCR. Results show relative expression value of PTEN- Δ or PTEN compared to control cells (pcDNA3 transfected cells). Significance was calculated by Student‘s T-test, * p < 0.05

Article Snippet: The human RCC cell lines A498, 786-O, Caki-1 and Caki-2 were obtained from LGC Promochem and CCF-RCI and CCF-RCII were kindly provided by the establisher, Cleveland Clinic Foundation [ ].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Construct, Western Blot, Positive Control, In Vitro, Control

Journal: Cell Communication and Signaling : CCS

Article Title: In renal cell carcinoma the PTEN splice variant PTEN-Δ shows similar function as the tumor suppressor PTEN itself

doi: 10.1186/s12964-018-0247-9

Figure Lengend Snippet: Influence of PTEN-Δ and PTEN on the functional behavior of renal tumor cells. a Migration of PTEN-Δ or PTEN transfected 786-O and A498 cells was determined in a Boyden chamber using ECM compounds as chemotaxins (FN: Fibronectin, VN: Vitronectin, LM: Laminin, CI: Collagen I and CIV: Collagen IV). Differences are shown in percentage of control cells (pcDNA3 transfected cells). b Cell adhesion of PTEN-Δ or PTEN transfected 786-O and A498 cells on immobilized ECM compounds were determined. BSA was used as control (Data not shown). Differences are shown in percentage of control cells (pcDNA3 transfected cells). c Apoptosis values of PTEN-Δ or PTEN transfected 786-O and A498 cells were quantified by determination of cytoplasmic histone-associated DNA fragments (Cell death detection assay, Roche). Differences are shown in percentage of control cells (pcDNA3 transfected cells). Significance was calculated by Student‘s T-test, * p < 0.05

Article Snippet: The human RCC cell lines A498, 786-O, Caki-1 and Caki-2 were obtained from LGC Promochem and CCF-RCI and CCF-RCII were kindly provided by the establisher, Cleveland Clinic Foundation [ ].

Techniques: Functional Assay, Migration, Transfection, Control, Detection Assay

Journal: Cell Communication and Signaling : CCS

Article Title: In renal cell carcinoma the PTEN splice variant PTEN-Δ shows similar function as the tumor suppressor PTEN itself

doi: 10.1186/s12964-018-0247-9

Figure Lengend Snippet: Influence of PTEN-Δ and PTEN on cell signaling. Expression of integrin α1 ( a ), integrin α5 ( b ) integrin αV ( c ) and integrin β1 ( d ) in RCC cells were determined in PTEN-Δ and PTEN transfected cells by flow cytometry. Representative histograms are shown for each staining. Expression value is shown as percentage of expression of the pcDNA3 transfected control cells. Significance was calculated by Student‘s T-test, * p < 0.05. Phosphorylation of AKT (T308) ( e ), AKT (S473) ( f ), p38 (T180/Y182) ( g ) and JNK (T183/Y185) ( h ) in transfected 786-O and A498 cells was determined by Western blot. Representative Western blots are shown for each kinase. Activity value was determined by densitometric evaluation and is shown as percentage of activity of the pcDNA3 transfected cells. Significance was calculated by Student‘s T-test, * p < 0.05

Article Snippet: The human RCC cell lines A498, 786-O, Caki-1 and Caki-2 were obtained from LGC Promochem and CCF-RCI and CCF-RCII were kindly provided by the establisher, Cleveland Clinic Foundation [ ].

Techniques: Expressing, Transfection, Flow Cytometry, Staining, Control, Phospho-proteomics, Western Blot, Activity Assay

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 was essential to cell proliferation. ( a ) The expression of ORM1 protein in A498, 786-O, and Caki-2 cells was much higher than the 293 T cells. ( b ) The gray value analysis of protein in ( a ). ( c ) ORM1 was knocked down in 786-O and Caki-2 cells. ( d ) The gray value analysis of protein in ( c ). ( e ) Cell proliferation of 786-O cells was inhibited in ORM1-KD group compared to NC group. ( f ) Cell proliferation of Caki-2 cells was inhibited in ORM1-KD group compared to NC group. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 was essential to cell migration and invasion. ( a ) Cell migration and invasion was suppressed in ORM1-KD group compared to NC group in 786-O and Caki-2 cells in transwell assay with/without Matrigel. ( b ) The statistical analysis of cells in cell migration in ( a ). ( c ) The statistical analysis of cells in cell invasion in a. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Migration, Transwell Assay

Journal: Scientific Reports

Article Title: ORM1 promotes tumor progression of kidney renal clear cell carcinoma (KIRC) through CALR-mediated apoptosis

doi: 10.1038/s41598-023-42962-w

Figure Lengend Snippet: ORM1 enhanced the efficiency of sorafenib in KIRC. ( a ) Sorafenib inhibited cell proliferation in concentration-dependent manner. ( b ) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in 786-O cells. ( c ) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in Caki-2 cells. # p < 0.05 showed statistically difference.

Article Snippet: KIRC cell lines including 786-O, A498, and Caki-2 cells were obtained from ICell Bioscience Inc, Shanghai (Shanghai, China).

Techniques: Concentration Assay