Evaluating Apoptosis Induction in Colon Cancer

Human colon carcinoma cell lines were treated, in duplicate, with equimolar concentrations (20 μM) of each agent that inhibits SMO (GDC-0449; JS Research), MEK (AZD6244; Selleckchem) or GLI (GANT61; Calbiochem). Following 72 hr exposure, cells were collected by trypsinization and incubated with Annexin V FITC (BD Biosciences, CA) and propidium iodide (Sigma, MO) prior to analysis using a FACSCalibur flow cytometer. Raw data were analyzed by CellQuest software [36 (link)-39 (link)].

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Agyeman A., Jha B.K., Mazumdar T, & Houghton J.A. (2014). Mode and specificity of binding of the small molecule GANT61 to GLI determines inhibition of GLI-DNA binding. Oncotarget, 5(12), 4492-4503.

Publication 2014

AZD6244 Annexin V FITC propidium iodide FACSCalibur flow cytometer

Annexin v fitc Azd6244 Carcinoma Cell lines Cells Colon Gant61 Gdc 0449 Human Inhibits Propidium iodide

Corresponding Organization :

Other organizations : Cleveland Clinic Lerner College of Medicine

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Variable analysis

independent variables

GDC-0449 (inhibits SMO)
AZD6244 (inhibits MEK)
GANT61 (inhibits GLI)

dependent variables

Annexin V FITC staining (apoptosis)
Propidium iodide staining (cell death)

control variables

Equimolar concentrations (20 μM) of each agent
Exposure time (72 hr)

controls

Duplicate treatments for each agent

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