Journal: PLoS ONE

Article Title: Identification of Tissue microRNAs Predictive of Sunitinib Activity in Patients with Metastatic Renal Cell Carcinoma

doi: 10.1371/journal.pone.0086263

Figure Lengend Snippet: Patient characteristics.

Article Snippet: A sunitinib-resistant Caki-2 cell line (SRCaki-2) was generated by continuous exposure for four months to gradually increasing doses of sunitinib: From 8 μM to 14 μM (Sutent, LC laboratories).

Techniques: Biomarker Discovery

Journal: PLoS ONE

Article Title: Identification of Tissue microRNAs Predictive of Sunitinib Activity in Patients with Metastatic Renal Cell Carcinoma

doi: 10.1371/journal.pone.0086263

Figure Lengend Snippet: , (sunitinib sensitive, n = 14 and sunitinib resistant, n = 6). MiR-942, miR-628-5p, miR-133a and miR-484 were overexpressed in sunitinib resistant patients. MiR-942 significantly discriminated between both patients groups. MiRNA levels are represented as 2 −ΔCt mean ± SEM. MiRNA expression was normalized against RNU6B. R = responders. NR = non responders.

Article Snippet: A sunitinib-resistant Caki-2 cell line (SRCaki-2) was generated by continuous exposure for four months to gradually increasing doses of sunitinib: From 8 μM to 14 μM (Sutent, LC laboratories).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Identification of Tissue microRNAs Predictive of Sunitinib Activity in Patients with Metastatic Renal Cell Carcinoma

doi: 10.1371/journal.pone.0086263

Figure Lengend Snippet: TTP and OS were significantly reduced in MRCC patients treated with sunitinib that had ( A ) miR-942 expression over the mean; ( B ) miR-628-5p levels over the mean; ( C ) miR-133a levels over p75; ( D ) and miR-484 expression over the mean.

Article Snippet: A sunitinib-resistant Caki-2 cell line (SRCaki-2) was generated by continuous exposure for four months to gradually increasing doses of sunitinib: From 8 μM to 14 μM (Sutent, LC laboratories).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Identification of Tissue microRNAs Predictive of Sunitinib Activity in Patients with Metastatic Renal Cell Carcinoma

doi: 10.1371/journal.pone.0086263

Figure Lengend Snippet: ( A ) Migration of HBMEC was significantly increased after co-culture with miR-942/Caki-2 (*: p<0.05), as compared with co-culture with miR-Neg/Caki-2 cells. ( B ) HBMEC became less sensitive to sunitinib activity after co-culture with miR-942/Caki-2 (LD 50 >35 µM) in comparison to HBMEC co-cultured with miR-Neg/Caki-2 cells (LD 50 13.5 µM). ( C ) Phosphorylation of VEGFR2 and p44/42 MAPK (Erk1/2) proteins was increased in HBMEC co-cultured with miR-942/Caki-2, as compared to the co-culture with miR-Neg/Caki-2 cells.

Article Snippet: A sunitinib-resistant Caki-2 cell line (SRCaki-2) was generated by continuous exposure for four months to gradually increasing doses of sunitinib: From 8 μM to 14 μM (Sutent, LC laboratories).

Techniques: Migration, Co-Culture Assay, Activity Assay, Comparison, Cell Culture, Phospho-proteomics

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