Journal: Oncotarget

Article Title: The novel combination of dual mTOR inhibitor AZD2014 and pan-PIM inhibitor AZD1208 inhibits growth in acute myeloid leukemia via HSF pathway suppression

doi:

Figure Lengend Snippet: A. PIM1 mRNA levels were analyzed in AML patient samples with FLT 3 wild-type (WT) or FLT 3 mutant (MT) from 2 studies (GSE14468 and GSE1159) by the Oncomine Platform. Error bars indicate SEM. B. PIM1, PIM2, and PIM3 protein levels in AML cell lines with FLT3 -WT (OCI-AML3, MOLM-16) or with FLT3 -ITD (MV4;11, MOLM-13, MOLM-14) were detected by Western blotting. The intensities compared to those of α-tubulin levels after background subtraction were obtained using ImageJ software.

Article Snippet: For immunoblotting, the following antibodies were used: alpha-tubulin and beta-actin (Sigma-Aldrich), PIM1 (Abcam, Cambridge, MA), PIM3 (Santa Cruz Biotechnology, Santa Cruz, CA), PIM2, p70 S6 Kinase, phospho-(p-) p70 S6 Kinase (Thr389), 4EBP1, phospho-(p-)4EBP1 (Thr37/46), AKT, p-AKT (Ser473), AMPK-α, pAMPK-α (Thr172), c-myc, GAPDH, and horseradish peroxidase-linked anti-mouse and anti-rabbit IgG (all from Cell Signaling Technology). iTRAQ sample labeling, mass spectrometry analysis, and peptide identification The iTRAQ chemical labeling mass spectrometry (MS) method was performed by following the manufacturer's protocol [ ].

Techniques: Mutagenesis, Western Blot, Software

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 1. Induction of c-myc and PIM1 expression in TCL cells (A) qRT-PCR was conducted to investigate PIM1 transcription in TCL cells. (B) qRT-PCR was con- ducted to investigate c-myc transcription in TCL cells. Results are presented as the mean ± SEM of three independent experiments. **P < 0.01 vs. PBMCs.

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Expressing, Quantitative RT-PCR

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 3. Doxorubicin-triggered apoptosis in TCL cells following PIM1 knockdown HUT-78 cells were transfected for 24 h with PIM1 siRNA and subsequently treated with 10 nM doxorubicin for 24 h. (A) Representative immunoblots and the quantitative evaluation of (B) PIM1 and (C) c-myc in HUT-78 cells. (D) Survival was evaluated by CCK-8 assay. (E) Quantification of cell apoptosis in each group by flow cytometry. Results are presented as the mean ± SEM of three inde- pendent experiments. *P < 0.05 and **P < 0.01 vs. control (Con).

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Knockdown, Transfection, Western Blot, CCK-8 Assay, Cytometry, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 2. Doxorubicin induces apoptosis via inhibition of PIM1-c-myc expression through the JAK-STAT3 pathway HUT-78 cells were treated with 10 nM doxorubicin for 24 h. (A) Representative immunoblots and the quantitative evaluation of (B) PIM1, (C) c-myc, (D) p-STAT3, and (E) p-JAK in HUT-78 cells. (F) Survival was evaluated by CCK-8 assay. (G) Quantification of cell apoptosis in each group by flow cytometry. Results are presented as the mean ± SEM of three independent experiments. *P < 0.05 and **P < 0.01 vs. control (Con).

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Inhibition, Expressing, Western Blot, CCK-8 Assay, Cytometry, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 4. Suppression of doxorubicin-triggered apoptosis in TCL cells by PIM1 overexpression HUT-78 cells were transfected for 24 h with PIM1 cDNA and subsequently treated with 10 nM doxorubicin for 24 h. (A) Representative immunoblots and the quantitative evaluation of (B) PIM1 and (C) c-myc in HUT-78 cells. (D) Survival was evaluated by CCK-8 assay. (E) Quantification of cell apoptosis in each group by flow cytometry. Results are presented as the mean ± SEM of three independent experiments. *P < 0.05 and **P < 0.01 vs. control (Con).

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Over Expression, Transfection, Western Blot, CCK-8 Assay, Cytometry, Control

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 6. Overview of the contribution of PIM1 to doxorubicin resistance via c-myc In TCL cells, the over-activated JAK-STAT3 pathway increased PIM1 expression and upregulated the expression of c-myc, which suppressed apoptosis and eventually led to doxorubicin resistance.

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: PIM1 overexpression in T-cell lymphomas protects tumor cells from apoptosis and confers doxorubicin resistance by upregulating c-myc expression.

doi: 10.1093/abbs/gmy076

Figure Lengend Snippet: Figure 5. Doxorubicin resistance is acquired through upregulation of c-myc expression in TCL cells via PIM1 overexpression HUT-78 cells were transfected for 24 h with c-myc siRNA and PIM1 cDNA and subsequently treated with 10 nM of doxorubicin for 24 h. (A) Representative immunoblots and the quantitative evaluation of (B) c-myc in HUT-78 cells. (C) Quantification of cell apoptosis in each group by flow cytometry. Results are presented as the mean ± SEM of three independent experiments. **P < 0.01 vs. control (Con). (D) Survival was evaluated by CCK-8 assay. (E) Quantification of cell apoptosis in each group by flow cytometry. Results are presented as the mean ± SEM of three independent experiments. *P < 0.05 and #P < 0.05.

Article Snippet: Knockdown of PIM1 or c-myc via siRNA PIM1 siRNA (target sequence: 5′-GGUGUGUGGAGAUAUUCCU TT-3′) and control siRNA (target sequence: 5′-UUCUCCGAACGU GUCACGUTT-3′), and c-Myc siRNA (target sequence: 5′-AACGUUA GCUUCACCAACAUU-3′) and control siRNA (target sequence: 5′-AAUUCUCCGAACGUGUCACGU-3′) (100 nM; Santa Cruz Biotechnology, Paso Robles, USA) were transiently transfected with RNAiFect Transfection Reagent (Qiagen, Hilden, Germany).

Techniques: Expressing, Over Expression, Transfection, Western Blot, Cytometry, Control, CCK-8 Assay