igf1r inhibitor osi 906  (MedChemExpress)

Journal: Theranostics

Article Title: Overexpression of BACH1 mediated by IGF2 facilitates hepatocellular carcinoma growth and metastasis via IGF1R and PTK2

doi: 10.7150/thno.65775

Figure Lengend Snippet: BACH1 regulates cell motility-associated genes IGF1R and PTK2 expression. (A) The overlap of up-regulated genes in PLC/PRF/5-LV-BACH1 cells VS PLC/PRF/5-LV-Control cells and down-regulated genes in MHCC97H-shBACH1 cells VS MHCC97H-shControl cells (fold change ≥ 2.0) were detected by the Human Cell Motility PCR Array and showed by Venn diagrams. (B-C) The effects of BACH1 overexpression or knockdown on IGF1R and PTK2 expression. The mRNA (B) and protein (C) levels of IGF1R and PTK2 in the PLC/PRF/5-LV-BACH1, MHCC97H-shBACH1 and corresponding control cells were determined by RT-qPCR and western blot. (D) Luciferase activities of IGF1R and PTK2 promoter reporter vectors in the PLC/PRF/5 cells cotransfected with pCMV-BACH1 or pCMV-Taq. (E-F) Luciferase activities of serially truncated/mutated IGF1R (E) and PTK2 (F) promoters reporter vectors in the PLC/PRF/5 cells cotransfected with pCMV-BACH1or pCMV-Taq. (G-J) ChIP analysis of BACH1 or RNA polymerase II (Pol II) occupancy in the IGF1R , PTK2 or CXCR4 promoter region in the indicated PLC/PRF/5 cells and HCC tissues (n = 6). An isotype-matched IgG was used as a negative control. CXCR4 promoter region and Pol II were used as positive controls. *P < 0.05, **P < 0.01, ***P < 0.001. Data were shown as Mean ± SD.

Article Snippet: PI3K inhibitor LY294002 (S1105), ERK inhibitor SCH772984 (S7101), IGF1R inhibitor linsitinib (OSI-906) (S1091), PTK2 inhibitor defactinib (VS-6063) (S7654), TAE226 (S2820) were purchased from Selleck Chemicals (Selleck, TX, USA).

Techniques: Expressing, Control, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Luciferase, Negative Control

Journal: Theranostics

Article Title: Overexpression of BACH1 mediated by IGF2 facilitates hepatocellular carcinoma growth and metastasis via IGF1R and PTK2

doi: 10.7150/thno.65775

Figure Lengend Snippet: BACH1 accelerates HCC growth and metastasis via directly upregulating IGF1R and PTK2. (A) IGF1R and PTK2 were knocked down in BACH1-overexpressing PLC/PRF/5 cells or overexpressed in BAHC1-knockdown MHCC-97H cells. The protein levels of BACH1, p-IGF1R, IGF1R, p-PTK2 and PTK2 in the indicated HCC cells were determined by western blot. (B-C) The migratory and invasive capacity of the indicated PLC/PRF/5 and MHCC97H cells were detected by transwell assay. Scale bar, 100 µm. (D-I) HCC orthotopic xenograft models indicated that BACH1 promotes HCC growth and metastasis by upregulating IGF1R and PTK2. The representative bioluminescent imaging in the liver (D), the bioluminescent signals of liver tumors (E), the occurrence of pulmonary metastasis (F), the representative H&E staining images of pulmonary tissues (G), the number of pulmonary metastasis lesions (H) from nude mice after orthotopic transplantation with indicated PLC/PRF/5 and MHCC97H cells were shown. The OS of mice was analyzed in (I). Scale bars, 2 mm (upper), 200 µm (lower). *p < 0.05, **p < 0.01, ***p < 0.001. Data were shown as Mean ± SD.

Article Snippet: PI3K inhibitor LY294002 (S1105), ERK inhibitor SCH772984 (S7101), IGF1R inhibitor linsitinib (OSI-906) (S1091), PTK2 inhibitor defactinib (VS-6063) (S7654), TAE226 (S2820) were purchased from Selleck Chemicals (Selleck, TX, USA).

Techniques: Knockdown, Western Blot, Transwell Assay, Imaging, Staining, Transplantation Assay

Journal: Theranostics

Article Title: Overexpression of BACH1 mediated by IGF2 facilitates hepatocellular carcinoma growth and metastasis via IGF1R and PTK2

doi: 10.7150/thno.65775

Figure Lengend Snippet: IGF2 promotes HCC growth and metastasis depending on BACH1. (A) BACH1 was silenced in PLC/PRF/5 cells. IGF1R and PTK2 were overexpressed in BACH1-silencing PLC/PRF/5 cells. These cells were treated with IGF2 at 100 ng/ml for 24 h. The protein levels of BACH1, p-IGF1R, IGF1R, p-PTK2 and PTK2 in the indicated PLC/PRF/5 cells were detected by western blot. (B-C) The migratory and invasive capacity of the indicated PLC/PRF/5 cells were analyzed by transwell assay. Scale bar, 100 µm. (D) IGF2 was overexpressed in PLC/PRF/5 cells and then BACH1 was silenced in IGF2-overexpressing PLC/PRF/5 cells. IGF1R and PTK2 were overexpressed in BACH1-silencing PLC/PRF/5-LV-IGF2 cells. The protein levels of IGF2, BACH1, p-IGF1R, IGF1R, p-PTK2 and PTK2 in the indicated PLC/PRF/5 cells were detected by western blot. (E-F) The migratory and invasive capacity of the indicated PLC/PRF/5 cells were detected by transwell assay. Scale bar, 100 µm. (G-L) The indicated HCC cells were used to construct HCC orthotopic xenograft models. The representative bioluminescent imaging in the liver (G), the bioluminescent signals of liver tumors (H), the occurrence of pulmonary metastasis (I), the representative H&E staining images of pulmonary tissues (J), the number of pulmonary metastasis lesions (K) from nude mice after orthotopic transplantation with indicated PLC/PRF/5 cells were shown. The OS of mice was analyzed in (L). Scale bars, 2 mm (upper), 200 µm (lower). *p < 0.05, **p < 0.01, ***p < 0.001. Data were shown as Mean ± SD.

Article Snippet: PI3K inhibitor LY294002 (S1105), ERK inhibitor SCH772984 (S7101), IGF1R inhibitor linsitinib (OSI-906) (S1091), PTK2 inhibitor defactinib (VS-6063) (S7654), TAE226 (S2820) were purchased from Selleck Chemicals (Selleck, TX, USA).

Techniques: Western Blot, Transwell Assay, Construct, Imaging, Staining, Transplantation Assay

Journal: Theranostics

Article Title: Overexpression of BACH1 mediated by IGF2 facilitates hepatocellular carcinoma growth and metastasis via IGF1R and PTK2

doi: 10.7150/thno.65775

Figure Lengend Snippet: The levels of BACH1 positively correlates with IGF1R and PTK2 levels in human HCC tissues. (A) Representative IHC staining of BACH1, IGF1R, and PTK2 in primary HCC tissues and matched nonneoplastic tissues were shown. Scale bars, 200 µm (upper), 50 µm (lower). (B) The correlation analysis of BACH1 and IGF1R or PTK2 levels in two HCC cohorts. (C-F) The correlations between IGF1R, PTK2, BACH1/IGF1R, or BACH1/PTK2 expression and OS or recurrence rates were analyzed by Kaplan-Meier. (G) Relative mRNA expression of BACH1 , IGF1R and PTK2 in adjacent nonneoplastic specimens (n = 20), primary HCC specimens (n = 20) and metastatic HCC specimens (n = 20) were analyzed by RT-qPCR. (H-I) Representative IHC staining (H) and IHC scores (I) of BACH1, IGF1R, and PTK2 in adjacent nonneoplastic specimens (n = 20), primary HCC specimens (n = 20), and metastatic HCC specimens (n = 20) were shown. Scale bars, 200 µm (left), 50 µm (right). *p < 0.05, **p < 0.01, ***p < 0.001. Data were shown as Mean ± SD.

Article Snippet: PI3K inhibitor LY294002 (S1105), ERK inhibitor SCH772984 (S7101), IGF1R inhibitor linsitinib (OSI-906) (S1091), PTK2 inhibitor defactinib (VS-6063) (S7654), TAE226 (S2820) were purchased from Selleck Chemicals (Selleck, TX, USA).

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR

Journal: Theranostics

Article Title: Overexpression of BACH1 mediated by IGF2 facilitates hepatocellular carcinoma growth and metastasis via IGF1R and PTK2

doi: 10.7150/thno.65775

Figure Lengend Snippet: IGF1R blockade combined with PTK2 inhibition suppresses BACH1-mediated HCC growth and metastasis. (A) The protein levels of BACH1, p-IGF1R, IGF1R, p-PTK2, PTK2 in PLC/PRF/5-LV-BACH1 cells treated with linsitinib alone or defactinib alone or a combination of both. (B-C) Transwell assay was performed to analyze the migratory and invasive capacity of PLC/PRF/5-LV-BACH1 cells treated with linsitinib alone or defactinib alone or a combination of both. Scale bar, 100 µm. (D) Schematic diagram of drug treatment for nude mice. (E-J) HCC orthotopic xenograft models indicated that the combination of linsitinib and defactinib suppressed BACH1-mediated HCC growth and metastasis. The representative bioluminescent imaging in the liver (E), the bioluminescent signals of liver tumors (F), the occurrence of pulmonary metastasis (G), the representative H&E staining images of pulmonary tissues (H), the number of pulmonary metastasis lesions (I) from nude mice after orthotopic transplantation with PLC/PRF/5-LV-BACH1 cells treated with linsitinib alone or defactinib alone or a combination of both were shown. The OS of mice was analyzed in (J). Scale bars, 2 mm (upper), 200 µm (lower). *p < 0.05, **p < 0.01, ***p < 0.001. Data were shown as Mean ± SD. (K) A schematic diagram showing how BACH1 facilitates HCC growth and metastasis and the combination strategy for HCC. BACH1 upregulates IGF1R and PTK2 to promote HCC growth and metastasis. IGF2, the ligand of IGF1R, in turn upregulates BACH1 expression through the IGF1R-ERK1/2-ETS1 cascades, thus forming a positive feedback loop to stimulate HCC cells continuously. IGF1R blockade combined with PTK2 inhibition significantly suppresses BACH1-mediated HCC malignant progression.

Article Snippet: PI3K inhibitor LY294002 (S1105), ERK inhibitor SCH772984 (S7101), IGF1R inhibitor linsitinib (OSI-906) (S1091), PTK2 inhibitor defactinib (VS-6063) (S7654), TAE226 (S2820) were purchased from Selleck Chemicals (Selleck, TX, USA).

Techniques: Inhibition, Transwell Assay, Imaging, Staining, Transplantation Assay, Expressing

Journal: Cells

Article Title: High Glucose Concentrations Negatively Regulate the IGF1R/Src/ERK Axis through the MicroRNA-9 in Colorectal Cancer

doi: 10.3390/cells8040326

Figure Lengend Snippet: High glucose (HG) concentrations induced epithelial-to-mesenchymal transition protein expression and enhanced migration activity in colorectal cancer (CRC) cells. SW480 (low metastatic potential) and SW620 (high metastatic potential) cells were cultured in different concentrations of glucose (normal: NG; HG; and osmotic control: NG + l -glucose). ( A ) Morphological change occurred from epithelial to mesenchymal type in the HG-concentration group. ( B ) HG concentration caused downregulation of E-cadherin and upregulation of N-cadherin, βCTN, and vimentin, but c-myc was unchanged, as detected using Western blotting. β-actin was evaluated as an internal control. ( C , D ) Wound healing assay showed that HG concentration promoted cell motility in SW480 and SW620 CRC cells after 48 and 72 h of culture, compared with the NG and NG + l -glucose groups. ( E ) In a Transwell migration assay, 3.5 × 10 5 SW480 and SW620 CRC cells were plated onto a 24-well plate and cultured in NG and HG-concentration medium for 96 h. HG concentration promoted cell motility in SW480 and SW620 cells. NG + l -glucose cells were evaluated as ostomic controls. ( F ) These data show that HG concentration caused upregulation of p-IGF1R in CRC. In addition, HG concentration promoted IGF1R downstream signaling, including p-Src and p-ERK; these proteins were increased when CRC cells were cultured in HG-concentration medium. Levels of β-actin were evaluated as loading controls. Statistically significant differences between the two groups were judged using Student’s t -tests; * p < 0.05, ** p < 0.005, *** p < 0.001; n.s. = nonsignificant.

Article Snippet: The IGF1R inhibitor OSI-906 was purchased from Selleckchem ® (#S1091), and the Src inhibitor PP1 was obtained from Calbiochem ® (#567809).

Techniques: Expressing, Migration, Activity Assay, Cell Culture, Control, Concentration Assay, Western Blot, Wound Healing Assay, Transwell Migration Assay

Journal: Cells

Article Title: High Glucose Concentrations Negatively Regulate the IGF1R/Src/ERK Axis through the MicroRNA-9 in Colorectal Cancer

doi: 10.3390/cells8040326

Figure Lengend Snippet: High glucose (HG) concentrations regulated IGF1R and Src and promoted the downstream signaling pathway in colorectal cancer (CRC) cells. ( A , B ) OSI-906 (IGF1R inhibitor) or ( C , D ) PP1 (Src inhibitor) affected proliferation in a dose-dependent manner in CRC cells. First, 3.5 × 10 5 SW480 and SW620 cells were seeded onto a 24-well plate. After incubation overnight, they were treated with OSI-906 (1.0 μM and 2.5 μM) or PP1 (2.0 μM and 4.0 μM). These data show that OSI-906 and PP1 significantly inhibited proliferation induced by HG concentration in SW480 and SW620 cells at 1.0 μM and 2.5 μM doses or 2.0 μM and 4.0 μM doses compared with the control group (dimethyl sulfoxide, DMSO). ( E – H ) Metastatic activities of CRC cells treated with OSI-906 or PP1 were detected using a Transwell assay; 3.5 × 10 5 SW480 and SW620 cells were plated onto a 24-well plate and incubated overnight after treatment with OSI-906 2.5 μM or PP1 2.0 μM for 96 h. These data show that OSI-906 (2.5 μM) and PP1 (2 μM) significantly inhibited the migration viability of SW480 and SW620 cells, which was promoted by HG concentration, compared with the HG-concentration group evaluated as positive controls. In addition, 3.5 × 10 5 SW480 or SW620 cells were cultured in basement membrane matrix-coated 24-well plates with HG-concentration medium and then treated with OSI-906 or PP1 for 168 h. These data show that OSI-906 and PP1 significantly inhibited the invasion viability of SW480 and SW620 cells. ( I , J ) Western blot analysis data suggest that OSI-906 or PP1 treatment reduced p-IGF1R or p-Src downstream signaling, reduced the expression of cell-cycle-regulated proteins, and induced and reduced expression of epithelial-to-mesenchymal transition proteins in HG-concentration medium compared with the control group (DMSO). HG concentration was evaluated as a positive control and levels of β-actin were evaluated as loading controls. These data are expressed as mean ± SEM and are representative of two independent experiments according to Student’s t -tests; * p < 0.05, ** p < 0.005; n.s. = nonsignificant.

Article Snippet: The IGF1R inhibitor OSI-906 was purchased from Selleckchem ® (#S1091), and the Src inhibitor PP1 was obtained from Calbiochem ® (#567809).

Techniques: Incubation, Concentration Assay, Control, Transwell Assay, Migration, Cell Culture, Membrane, Western Blot, Expressing, Positive Control

Journal: Cells

Article Title: High Glucose Concentrations Negatively Regulate the IGF1R/Src/ERK Axis through the MicroRNA-9 in Colorectal Cancer

doi: 10.3390/cells8040326

Figure Lengend Snippet: Expression and regulation of miR-9 in colorectal cancer (CRC) cell lines by high glucose (HG) concentration. TaqMan quantitative real-time polymerase chain reaction analysis was performed on miR-9 in SW480 and SW620 CRC cell lines that were cultured in different concentrations of glucose: Namely NG (5.5 mM d -glucose) and HG (25 mM d -glucose). In HG-concentration medium, miR-9 was decreased in both ( A ) SW480 ( p < 0.005) and ( B ) SW620 ( p < 0.05). All data were analyzed using a relative quantification method (2 −∆∆Ct ) with RNU6B small RNA as an internal control. ( C ) SW480 and ( D ) SW620 cells were transfected with pre-miR-9 at different doses (15 and 30 nM) or pre-miR negative control (NC) for 48 h in HG-concentration medium. ( E ) Western blotting validated the downregulation of p-IGF1R, cyclin B1, and N-cadherin as well as the upregulation of E-cadherin expression through pre-miR-9 overexpression, compared with pre-miR NC. β-actin was evaluated as an internal control. Statistically significant differences between the two groups were judged using Student’s t -tests, * p < 0.05, ** p < 0.005, *** p < 0.001; n.s. = nonsignificant.

Article Snippet: The IGF1R inhibitor OSI-906 was purchased from Selleckchem ® (#S1091), and the Src inhibitor PP1 was obtained from Calbiochem ® (#567809).

Techniques: Expressing, Concentration Assay, Real-time Polymerase Chain Reaction, Cell Culture, Quantitative Proteomics, Control, Transfection, Negative Control, Western Blot, Over Expression

Journal: Cells

Article Title: High Glucose Concentrations Negatively Regulate the IGF1R/Src/ERK Axis through the MicroRNA-9 in Colorectal Cancer

doi: 10.3390/cells8040326

Figure Lengend Snippet: Molecular mechanism through which high glucose (HG) concentration promotes proliferation and migration in colorectal cancer (CRC) cells. HG concentration activated pIGF1R and p-Src expression and increased downstream signaling by mediating the downregulation of miR-9 expression. Moreover, OSI-906 decreased the expression of the EMT protein N-cadherin and reduced the expression of the cell-cycle-regulated protein cyclin B1, as determined through Western blotting, but only cyclin B1 and E-cadherin were unchanged in SW620 cells ( I). Similarly, PP1 decreased the expression of the EMT protein N-cadherin and reduced the expression of the cell-cycle-regulated protein cyclin B1, as determined through Western blotting ( J), compared with the control group (dimethyl sulfoxide) cultured in HG-concentration medium. These data demonstrate that HG concentration promoted CRC cell proliferation, modulated EMT protein expression and morphology, and promoted cell migration and invasion ability through the IGF1R/Src/ERK pathway. In addition, miR-9-transfected cells expressed lower levels of p-IGF1R, cyclin B1, and N-cadherin, but E-cadherin was more upregulated compared with the negative control-transfected SW480 and SW620 cells, as determined through Western blotting. Thus, miR-9 is a tumor-suppressive microRNA that may regulate through the IGF1R/Src/ERK pathway the targeting of cyclin B1, N-cadherin, and E-cadherin in CRC cells in an HG-concentration environment ( E).

Article Snippet: The IGF1R inhibitor OSI-906 was purchased from Selleckchem ® (#S1091), and the Src inhibitor PP1 was obtained from Calbiochem ® (#567809).

Techniques: Concentration Assay, Migration, Expressing, Western Blot, Control, Cell Culture, Transfection, Negative Control

Journal: BMC Cancer

Article Title: No preclinical rationale for IGF1R directed therapy in chondrosarcoma of bone

doi: 10.1186/s12885-016-2522-8

Figure Lengend Snippet: a-b qRT-PCR and western blot analyses, respectively, reveal heterogeneous expression of IGF pathway members in chondrosarcoma cell lines. c Immunoprecipitation with IRS1 followed by a western blot for phospho-IRS1 reveals pathway activity in two out of three chondrosarcoma cell lines tested. d Evaluation of IGF1R downstream targets reveals an effect of OSI-906 on IGF1R and pAkt but not on pS6 and disphosphorylated ERK-1&2. Cell lines were treated for 72 hours with DMSO, 1 μM OSI-906 and/or 50 ng/ml IGF1

Article Snippet: The IGF1R inhibitors OSI-906, NVP-ADW742 and GSK1838705A were purchased from Selleck Chemicals LCC (see Additional file : Table S1 for properties) and dissolved in DMSO in a concentration of 10 mM.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunoprecipitation, Activity Assay

Journal: BMC Cancer

Article Title: No preclinical rationale for IGF1R directed therapy in chondrosarcoma of bone

doi: 10.1186/s12885-016-2522-8

Figure Lengend Snippet: Relative cell viability of 72 hours of treatment with IGF1R/IR inhibitors. a - b OSI-906 does not inhibit chondrosarcoma cell viability, even in the presence of IGF1. c - d IGF1R inhibitors NVP-ADW742 and GSK1838705A do not inhibit chondrosarcoma cell viability

Article Snippet: The IGF1R inhibitors OSI-906, NVP-ADW742 and GSK1838705A were purchased from Selleck Chemicals LCC (see Additional file : Table S1 for properties) and dissolved in DMSO in a concentration of 10 mM.

Techniques:

Journal: BMC Cancer

Article Title: No preclinical rationale for IGF1R directed therapy in chondrosarcoma of bone

doi: 10.1186/s12885-016-2522-8

Figure Lengend Snippet: IGF1R expression is high in chondrosarcoma cell lines but low in primary tumours. a IGF1R expression in chondrosarcoma cell line JJ012. b and d IGF1R expression in L835 cell line and primary tumour, respectively. c a conventional chondrosarcoma sample that was classified as weak IGF1R staining. Black bars represent 50 μm

Article Snippet: The IGF1R inhibitors OSI-906, NVP-ADW742 and GSK1838705A were purchased from Selleck Chemicals LCC (see Additional file : Table S1 for properties) and dissolved in DMSO in a concentration of 10 mM.

Techniques: Expressing, Staining

Journal: BMC Cancer

Article Title: No preclinical rationale for IGF1R directed therapy in chondrosarcoma of bone

doi: 10.1186/s12885-016-2522-8

Figure Lengend Snippet: Immunohistochemistry demonstrates only weak IGF1R expression in uncultured cartilage tumours

Article Snippet: The IGF1R inhibitors OSI-906, NVP-ADW742 and GSK1838705A were purchased from Selleck Chemicals LCC (see Additional file : Table S1 for properties) and dissolved in DMSO in a concentration of 10 mM.

Techniques: Immunohistochemistry, Expressing

Journal: Frontiers in Oncology

Article Title: Succinate Dehydrogenase Deficiency in Pediatric and Adult Gastrointestinal Stromal Tumors

doi: 10.3389/fonc.2013.00117

Figure Lengend Snippet: Immunohistochemical analysis of KIT, IGF1R, SDHB, and SDHA expression in WT and KIT mutant GISTs . Primary antibodies used include KIT (Dako), IGF1R (Cell Signaling), SDHB (Abcam), and SDHA (Abcam). Positive KIT staining is evident throughout tumor tissue in all cases (A,E,I,M) . Strong staining for IGF1R is seen in the WT GISTs (B,F,J) but not in the KIT mutant GIST (N) . SDHB staining is evident in the mutant GIST (O) and in the adjacent normal tissue and epithelial cells in the WT cases, but absent in the tumor tissue (C,G,K) . SDHA staining is absent in an SDHB-deficient GIST with a truncating SDHA mutation (D) . Positive SDHA staining is evident in an SDHB-deficient GIST harboring compound heterozygous missense SDHA mutations (H) , and in an SDHB-deficient GIST with no identified SDH mutations (L) as well as in the mutant GIST (P) . See text for more detailed mutation descriptions. GIST cases have been previously reported (Belinsky et al., ): (A–D) , case 2; (E–H) , case 1; (I–L) , case 10; (M–P) , case 21.

Article Snippet: An ongoing phase II trial is testing the IGF1R inhibitor linsitinib (OSI-906, Astellas, NCT01560260) in GIST patients that lack mutations in KIT , PDGFRA , and BRAF .

Techniques: Immunohistochemical staining, Expressing, Mutagenesis, Staining