Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: The inhibitory activity of Jac-A on tumour cells via MTT Assay (IC 50 , μ M, n = 4, mean ± SD)

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Activity Assay, MTT Assay

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A caused K562 cell apoptosis. 10 6 of K562 cells were treated with different concentrations of Jac-A for 48 h. Cells were then stained with Annexin V-FITC and propidium iodide, and analysed using flow cytometry. Four different cell populations marked as the following: live cell population (PI - AV-), early apoptosis (PI - AV+), late apoptosis (PI + AV+) and dead cells (PI + AV-). Cells were treated with no Jac-A (A) , 0.5% DMSO (B) , or 0.1, 1, 5, 10 μM/L Jac-A (C, D, E and F) . The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Staining, Flow Cytometry

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A induced caspase-dependent apoptosis in K562 cells. (A) Cyt c release in Jac-A-induced apoptosis of K562 cells. 10 6 of K562 cells were treated with different concentration of Jac-A for 48 h. Cytosol and mitochondrial heavy membrane samples were prepared and subjected to immunoblot with anti-Cyt c specific antibodie as described in Materials and methods. The results are one representative of three independent experiments. (B) Western blot showing conspicuous cleavage of caspase-3, caspase-9, and PARP in K562 cells treated with Jac-A. K562 cells subjected to protein extract preparation after treated with 0 (control), 3, 6, 12 μM/L Jac-A for 48 h. Then, fifty micrograms extracted protein subjected to Western blot using anti-PARP, PARP, cleaved caspase-9, caspase-9, cleaved caspase-3, caspase-3, and β-actin. The results are one representative of three independent experiments. (C) Jac-A-induced apoptosis was inhibited in a concentration dependent manner by the Z-VAD-fmk. K562 cells were first treated with or without different concentrations of Z-VAD-fmk for 4 h, followed by the treatment of Jac-A (10 μM) for 48 h.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Concentration Assay, Membrane, Western Blot, Control

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A inhibited the heterodimerization of antiapoptotic proteins with pro-apoptotic proteins. Jac-A inhibited the binding between Bax with Bcl-x L or Bcl-2 (A) and Bak with Mcl-1 (B) . K562 cells were treated with indicated concentrations of Jac-A for 48 h. 150 μg of K562 cell lysates were subjected to co-immunoprecipitation using anti-Bax and anti-Bak antibodies, respectively, further immunoblot with anti- Bcl-x L , Bcl-2, Mcl-1, Bax, or Bak antibody, as described in Materials and methods. The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Binding Assay, Immunoprecipitation, Western Blot

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Therapeutic study of Jac-A in the K562-bearing mice (n = 10/group). (A) Tumour volume plot of K562-bearing mice treated with vehicle or Jac-A at 2, 10, or 50 mg/kg by oral gavage for 21 days. The tumours were measured twice per week. The data are represented as the mean ± SEM. Tumour growth was inhibited significantly after treatment with Jac-A compared with the control group. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group. (B) Selected nude mice models of different groups treated with Jac-A or vehicle at day 14 after therapy. (C) Sizes of selected tumours harvested from dead nude mice bearing K562 cells from different groups treated with the vehicle or Jac-A. (D) Kaplan-Meier survival plot of the K562-bearing nude mice. The survival of the K562-bearing nude mice was prolonged in the Jac-A treated groups compared with control group. (E) Body weight plot of the K562-bearing nude mice. The data are represented as the mean ± SEM. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Control