Agents and methods for treating and preventing seborrheic keratosis
Provided herein are methods and assays for isolating and culturing seborrheic keratosis cells ex vivo. Also provided herein are screening assays using cultured seborrheic keratosis cells and methods for treating seborrheic keratosis in a subject.
1. A method for screening a candidate agent for inducing apoptosis, the method comprising:
(a) contacting a seborrheic keratosis cell or population of seborrheic keratosis cells with a candidate agent, and
(b) measuring apoptosis in the cell or population of cells, wherein an increase in apoptosis in the cell or population of cells indicates that the candidate agent induces apoptosis.
2. The method of claim 1 , wherein the candidate agent comprises an Akt signaling pathway inhibitor.
3. The method of claim 1 , wherein the seborrheic keratosis cell or population of seborrheic keratosis cells are cultured using a method comprising the steps of:
(a) contacting a biological sample comprising seborrheic keratosis cells obtained from a subject with a solution comprising a dispase enzyme at a temperature and for a time sufficient to initiate dissociation of the seborrheic keratosis cells from the biological sample, and
(b) culturing the dissociated seborrheic keratosis cells.
4. The method of claim 1 , wherein apoptosis is measured using sulforhodamine B (SRB) assay, MTT tetrazolium dye, TUNEL staining, Annexin V staining, propidium iodide staining, DNA laddering, PARP cleavage, caspase activation, and/or assessment of cellular and nuclear morphology.
5. The method of claim 1 , wherein the candidate agent is a small molecule, a peptide inhibitor, or an RNAi molecule.
6. An assay comprising:
(a) contacting a population of dissociated seborrheic keratosis cells with a candidate agent,
(b) contacting the cells of step (a) with a detectable antibody specific for an apoptotic protein,
(c) measuring the intensity of the signal from the bound, detectable antibody,
(d) comparing the measured intensity of the signal with a reference value and if the measured intensity is increased relative to the reference value,
(e) identifying the candidate agent as an inducer of apoptosis in the cell.
7. The assay of claim 6 , wherein the candidate agent comprises an Akt signaling pathway inhibitor.
8. The assay of claim 6 , wherein the population of seborrheic keratosis cells is cultured using a method comprising the steps of:
(a) contacting a biological sample comprising seborrheic keratosis cells obtained from a subject with a solution comprising a dispase enzyme at a temperature and for a time sufficient to initiate dissociation of the seborrheic keratosis cells from the biological sample, and
(b) culturing the dissociated seborrheic keratosis cells.
9. The assay of claim 6 , wherein the apoptotic protein is a caspase protein, a PARP protein, or a cleavage product thereof.