Rational Design of Human Prolactin Receptor Antagonists for Breast Cancer Therapy.

摘要

There were three tasks proposed for the second year of this project. Majority of the tasks have been accomplished. We were able to produce and purify large amount of hPRl and hPRL-G129R using E. coli expression system. The purified proteins were used to carry out in vitro as well as in vivo characterization of the hPRL antagonist (four manuscripts and six abstracts were published) . We further confirmed that the antagonistic effects of hPRL-Gl29R in breast cancer cells are probably through the inhibition of STATs phosphorylation, caspase activation, or TGFs modulation. Based upon our initial studies, we have found that that proposed multi-cell receptor level comparison using conventional binding assay was not sensitive enough for differentiate the difference. We carried out real time RT-PCR studies to quantify the PRL-receptor level (manuscript published) . One negative result is also included in this report. We were unable to produce hPRL-BP in E.coli expression system. We are trying to use a new strain of E. coli from Novogen to overcome this problem in the coming year.