Screening for Inhibitors of Prostaglandin E Synthase 3 Useful for Treatment of Prostate Cancer
AssigneeThe Regents of the University of California
InventorsLuke Gilbert, Felix Feng, Haolong Li
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Abstract
Methods and compositions are provided for screening candidate agents for inhibition of prostaglandin E synthase 3 (PTGES3) and anti-cancer activity against prostate cancer. In one aspect, a method of screening for a prostaglandin E synthase 3 (PTGES3) inhibitor for treating prostate cancer is provided, the method comprising: a) contacting PTGES3 with a candidate agent; and b) measuring inhibition of PTGES3 activity by the candidate agent. Screening assays may include determining the effectiveness of candidate PTGES3 inhibitors in reducing proliferation, survival, or androgen receptor abundance of prostate cancer cells.
Claims26
1-15. (canceled)
An androgen receptor-fluorescent protein reporter system comprising:
- a prostate cancer cell expressing a split fluorescent protein comprising a first component of a fluorescent protein and a second component of the fluorescent protein, wherein the prostate cancer cell comprises a modified genome comprising a first nucleic acid comprising a coding sequence encoding the first component of the fluorescent protein connected to a 5′ end of a protein coding exon of an androgen receptor (AR) gene such that the prostate cancer cell expresses a fusion protein comprising the first component of the fluorescent protein linked to the N-terminus of the androgen receptor;
- and
- a vector comprising an expression cassette comprising a second nucleic acid comprising a coding sequence encoding the second component of the fluorescent protein, wherein a reconstituted fluorescent protein forms when the first component and the second component of the split fluorescent protein are expressed and interact to produce a detectable fluorescent signal indicative of the level of the androgen receptor in the prostate cancer cell.
A method of determining localization or measuring a level of an androgen receptor in a cell, the method comprising:
- providing a prostate cancer cell expressing a split fluorescent protein comprising a first component of a fluorescent protein and a second component of the fluorescent protein, wherein the cell comprises a modified genome comprising a first nucleic acid comprising a coding sequence encoding the first component of the fluorescent protein connected to a 5′ end of a protein coding exon of an androgen receptor (AR) gene such that the prostate cancer cell expresses a fusion protein comprising the first component of the fluorescent protein linked to the N-terminus of the androgen receptor;
- transfecting the prostate cancer cell with a vector comprising an expression cassette comprising a second nucleic acid comprising a coding sequence encoding the second component of the fluorescent protein, wherein a reconstituted fluorescent protein forms when the first component and the second component of the split fluorescent protein are expressed and interact to produce a detectable fluorescent signal;
- and
- detecting the fluorescent signal, wherein the level or localization of the androgen receptor protein in the cell is determined from the fluorescent signal.
Dependencies are inferred from the claim text.