Journal: Iranian Journal of Basic Medical Sciences

Article Title: Quinazoline derivative compound (11d) as a novel angiogenesis inhibitor inhibiting VEGFR2 and blocking VEGFR2-mediated Akt/mTOR /p70s6k signaling pathway

doi:

Figure Lengend Snippet: Inhibition of VEGFR2 kinase activity by quinazoline derivative 11d and SU6668 was analyzed using an in vitro HTScan® VEGF receptor 2 kinase kit (Cell Signaling Technology, Danvers, MA, USA) combined with colorimetric ELISA detection according to the manufacturer’s instructions. Values are mean ± SEM (n = 6) of three independent experiments

Article Snippet: In vitro VEGFR2 kinase inhibition assay was performed using recombinant human VEGFR2 (Sino Biological Inc., USA) and VEGFR2 kinase assay Kit (GENMED SCIENTIFICS INC., USA).

Techniques: Inhibition, Activity Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Quinazoline derivative compound (11d) as a novel angiogenesis inhibitor inhibiting VEGFR2 and blocking VEGFR2-mediated Akt/mTOR /p70s6k signaling pathway

doi:

Figure Lengend Snippet: mRNA expression of VEGF and VEGFR2. Compound 11d reduced the mRNA expression of VEGFA and VEGFR2 in a dosedependent manner. HUVECs were treated with increasing concentrations of compound 11d for 24 hr

Article Snippet: In vitro VEGFR2 kinase inhibition assay was performed using recombinant human VEGFR2 (Sino Biological Inc., USA) and VEGFR2 kinase assay Kit (GENMED SCIENTIFICS INC., USA).

Techniques: Expressing

Journal: Iranian Journal of Basic Medical Sciences

Article Title: Quinazoline derivative compound (11d) as a novel angiogenesis inhibitor inhibiting VEGFR2 and blocking VEGFR2-mediated Akt/mTOR /p70s6k signaling pathway

doi:

Figure Lengend Snippet: (A) Compound 11d inhibited HepG-2 cell growth. Values are expressed as mean ± SEM ( n = 6) of three independent experiments; P < 0.05 versus vehicle control. (B) Compound 11d inhibited the VEGFR2-mediated AKT/mTOR/P70S6K pathway in HCC cells

Article Snippet: In vitro VEGFR2 kinase inhibition assay was performed using recombinant human VEGFR2 (Sino Biological Inc., USA) and VEGFR2 kinase assay Kit (GENMED SCIENTIFICS INC., USA).

Techniques:

Journal: Cell Death & Disease

Article Title: A novel angiogenesis inhibitor impairs lovo cell survival via targeting against human VEGFR and its signaling pathway of phosphorylation

doi: 10.1038/cddis.2012.145

Figure Lengend Snippet: HMQ18–22 inhibited cell viability and decreased phosphorylation of VEGFR2, VEGFR1, Akt, PKC α and PLC γ -1 involved in angiogenesis. ( a ) HMQ18–22 decreased cell survival in lovo and HUVEC cells. ( b ) The AlphaScreen signal indicated HMQ18–22 decreased VEGFR phosphorylation. ( c ) Cell were treated with VEGF (50 ng/ml) for 30 min before extracting proteins with RIPA lysis buffer. HMQ18–22 decreased the phosphorylation of VEGFR2(Tyr 1214 ), VEGFR1(Tyr 1333 ), Akt(Tyr 326 ), PKC α (Tyr 657 ) and PLC γ -1(Tyr 771 ) by western blot analysis. On the contrary, the Raf1(Tyr 341 ) phosphorylation was not altered by HMQ18–22. Results were quantified by densitometry analysis of the bands form and then normalization to GAPDH protein. ( d ) Effect of HMQ18–22 on cells transfected with siRNAs targeting of VEGFR2, VEGFR1, Akt, PKC α or PLC γ -1. Quantification of RT-PCR data showed knockdown of VEGFR2, VEGFR1, Akt, PKC α and PLC γ -1; the bottom right panel showed the effect of HMQ18–22 on cells proliferation was attenuated in knockdown cells. Data were expressed as mean values±S.D. ( n =3). * P <0.05, ** P <0.01 versus the untreated control group.

Article Snippet: HTRF VEGFR2 kinase kit was purchased from Cisbio (Codolet, France).

Techniques: Amplified Luminescent Proximity Homogenous Assay, Lysis, Western Blot, Transfection, Reverse Transcription Polymerase Chain Reaction

Journal: Cell Death & Disease

Article Title: A novel angiogenesis inhibitor impairs lovo cell survival via targeting against human VEGFR and its signaling pathway of phosphorylation

doi: 10.1038/cddis.2012.145

Figure Lengend Snippet: HMQ18–22 inhibited tumor growth in nude mice bearing human colon cancer xenografts. ( a ) The representative xenografts of lovo human colon cancer in mice. ( b ) HMQ18–22 decreased the phosphorylation of VEGFR2(Tyr 1214 ), VEGFR1(Tyr 1333 ), Akt(Tyr 326 ), PKC α (Tyr 657 ) and PLC γ -1(Tyr 771 ) in the tumor tissues by western blot analysis. ( c ) Quantitation data of ( b ). Data were expressed as mean values±S.D. ( n =3). ** P <0.01 versus the untreated control

Article Snippet: HTRF VEGFR2 kinase kit was purchased from Cisbio (Codolet, France).

Techniques: Western Blot, Quantitation Assay