E2F REPORTER MELANOMA CELLS

摘要

A modified melanoma cell line capable of quantification of the effects of MEK inhibitors and CDK4/6 inhibitors in a quantitative, temporal and non-invasive manner both in vitro and in vivo.

主权项

1. A method for determining the efficacy of a compound of interest to inhibit E2F activity—comprising:
a. applying a compound of interest to a modified cell, wherein said melanoma cells harboring either BRAF/NRAS mutations or neither mutations involving a modified E2F reporter system to monitor the efficacy of CDK4/6 inhibitor-based therapies, wherein tumor cells are transduced with tdTomato fluorescent protein and an EGFP-firefly luciferase fusion gene under the control of E2F transcriptional response elements; b. determining the amount of tdTomato activity and determining the amount of firefly luciferase activity in the modified cell, and c. comparing the amount of tdTomato to the amount of firefly luciferase expression, to quantitate E2F reporter activity to tumor size, wherein tdTomato measures tumor size and firefly luciferase measures pathway activity in the tumor.