Journal: Oncology Letters

Article Title: Association between the expression of inhibitor of growth family member 4 and the progression of clear cell renal carcinoma

doi: 10.3892/ol.2017.6450

Figure Lengend Snippet: ING4 mRNA is significantly decreased in CCRC tissues when compared with normal renal tissues. (A) A total of 40 paired tumor and adjacent normal renal tissues were analyzed for ING4 mRNA level by reverse transcription-quantitative polymerase chain reaction. β-actin was used as an internal control. (B) Analysis of relative mRNA level (compared with normal renal tissue) according to Fuhrman nuclear grade (24) (grade I, n=7; grade II, n=16; grade 3, n=13; grade 4, n=4), lymphatic metastasis (n0, n=29; n1, n=7; n2, n=4), tumor clinical stage (stage I, n=9; stage II, n=12; stage III, n=14; stage IV, n=5). ***P<0.001 by t-test. CCRC, clear cell renal carcinoma; ING4, inhibitor of growth family member 4.

Article Snippet: An enhanced chemiluminescence detection system (Shanghai Jiapeng Technology Co., Ltd., Shanghai, China) was used to visualize the protein bands. β-actin (1:1,000; cat no. ab8226; Abcam, Cambridge, MA, USA) was used as an internal control.

Techniques: Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Association between the expression of inhibitor of growth family member 4 and the progression of clear cell renal carcinoma

doi: 10.3892/ol.2017.6450

Figure Lengend Snippet: ING4 expression is significantly decreased in CCRC tissues when compared with normal renal tissues. (A) A total of 40 paired tumor and adjacent normal renal tissues were analyzed for ING4 expression using western blotting. Decreased ING4 expression was observed in CCRC tissues compared with adjacent normal renal tissues. β-actin was used as internal control. (B) Average ING4 expression in 40 paired tumor and normal renal tissues samples. (C) Analysis of the expression level of ING4 protein (compared with normal renal tissue) according to tumor nuclear grade (grade I, n=7; grade II, n=16; grade III, n=13; grade IV, n=4), lymphatic metastasis (n0, n=29; n1, n=7; n2, n=4), tumor clinical stage (stage I, n=9; stage II, n=12; stage III, n=14; stage IV n=5). ***P<0.001 by t-test. CCRC, clear cell renal carcinoma; ING4, inhibitor of growth family member 4; NT, normal tissue.

Article Snippet: An enhanced chemiluminescence detection system (Shanghai Jiapeng Technology Co., Ltd., Shanghai, China) was used to visualize the protein bands. β-actin (1:1,000; cat no. ab8226; Abcam, Cambridge, MA, USA) was used as an internal control.

Techniques: Expressing, Western Blot

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 1. Expression of NF1 and dopamine receptors in MPNST cells by RT-PCR analysis. Amplification products obtained using specific primers which recognized NF1, APP, D1R, D2R, D3R, D4R and D5R generated bands of the expected length (Table I). Ribosomal protein S18 was used as a control in each PCR amplification. A 100-bp DNA ladder is shown on the left side of each gel (lane M) with bands labelled in bp units.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Generated, Control

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 2. Dose-response effects of 7-OH-PIPAT treatment on cell viability. Induction of cell viability in serum-starved MPNST cells treated with increasing concentrations of the D3R agonist after 24 (◆) and 48 h (∫). Cells cultured in presence of 10% FBS (control), in total absence of serum (SS), additioned with 0.1% DMSO (SS + vehicle) or 7-OH-PIPAT (SS + D3R agonist) were used for MTT measurements as described in Materials and methods. Values are expressed as mean ODs (n=6) ± SEM. Results are representative of at least three independent experiments. #p<0.05, ##p<0.01 or ###p<0.001 as compared to serum-starved cells treated with vehicle.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Cell Culture, Control

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 3. Dose-response effects of 7-OH-PIPAT treatment on oligonucleo- some formation. Inhibition of apoptosis in serum-starved MPNST cells treated with increasing concentrations of the D3R agonist after 24 (A) and 48 h (B). Cells cultured in presence of 10% FBS (control), in total absence of serum (SS), added with 0.1% DMSO (SS + vehicle) or 7-OH-PIPAT (SS + D3R agonist) were used for oligonucleosome formation measurements as described in Materials and methods. Values are expressed as mean ODs (n=6) ± SEM. The bar graph shows the results of three independent experiments. #p<0.05 or ###p<0.001 as compared to serum-starved cells treated with vehicle. ***p<0.001 as compared to control groups.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Inhibition, Cell Culture, Control

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 4. Hoechst 33258 nuclear staining of serum-starved MPNST cells treated with 7-OH-PIPAT. Representative photomicrographs of MPNST cells cultured with 10% serum (control), serum-starved (SS), 0.1% DMSO (SS + vehicle) or additioned with the indicated concentrations of 7-OH- PIPAT (SS + D3R agonist) after 48 h exposure. Cells were stained with the fluorescent nuclear dye Hoechst 33258 and viewed at x40 magnification. Hoechst 33258 binds to nuclear DNA and emits an intense fluorescence in apoptotic cells due to nuclear condensation and/or chromatin frag- mentation. Each condition was reproduced in three dishes per experiment. Representative photomicrographs of both apoptotic and normal cells were taken from three fields per dish in a fixed pattern.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Staining, Cell Culture, Control, Fluorescence

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 5. Quantitative real-time PCR and Western blot analyses of NF1 and neurofibromin expression in serum-starved MPNST cells treated with 7-OH- PIPAT. Results are presented as mean fold-changes of 10% FBS-cultured MPNST cells (control, n=4) ± SEM. Relative fold-changes of the NF1 gene after treatment with the indicated concentrations of the D3R agonist after 24 (A) and 48 h (B) were normalized to the endogenous ribosomal protein S18 (house-keeping gene) and then calculated using the comparative ΔCt method (20). Baseline expression levels of the control groups were set to 1. (##p<0.01 or ###p<0.001 as compared to serum-starved cells treated with vehicle; ***p<0.001 as compared to control groups). Representative neurofibromin immunoblots (C and D) obtained using 50 μg of homogenates from MPNST cells cultured under the same experimental conditions for mRNA measurements showed comparable changes in neurofibromin expression both after 24 (C) and 48 h (D). ß-tubulin was used as loading control in each experiment. The diagrams show the results of three independent experiments.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Cell Culture, Control

Journal: International journal of oncology

Article Title: Protective effect of the dopamine D(3) receptor agonist (7-OH-PIPAT) against apoptosis in malignant peripheral nerve sheath tumor (MPNST) cells.

doi: 10.3892/ijo_00000743

Figure Lengend Snippet: Figure 6. Immunofluorescence analysis of neurofibromin in serum-starved MPNST cells treated with 7-OH-PIPAT. Representative photomicrographs showing neurofibromin immunolocalization in MPNST cells cultured with 10% serum (control), serum-starved (SS), 0.1% DMSO (SS + vehicle) or additioned with the maximum concentration used (10-5 M) of the D3R agonist (SS + D3R agonist) after 48 h exposure. Photomicrographs shown are representative results taken from ten different fields from randomly selected slides. Magnification x20.

Article Snippet: ––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––– sodium dodecyl sulphate (SDS) protein gel loading solution (Invitrogen), boiled for 5 min, separated on 4-12% Bis-tris gel (Invitrogen) by electrophoresis and processed as previously described (22).

Techniques: Immunofluorescence, Cell Culture, Control, Concentration Assay