(171d) 3D Reporter Gene Assay for High-Throughput Drug Screening

Traditional cell-based assay has high probability of failure to identify effective and safe compounds in early drug discovery. Approaches to address this problem include engineering of cancer cell lines to indicate the drug mechanism of action, and applying of 3D cancer models to simulate the cancer tissue physiology. In this research, cancer cell lines MCF7, Panc-1 and A549 are engineered as reporter cells and are cultured on polyethylene terephthalate (PET)-based 3D scaffold for high throughput drug screening. The assay reports the regulation of two prominent cancer biomarkers, survivin and human telomerase reverse transcriptase (hTERT), which are silenced in normal adult cells but are overexpressed in more than 85% tumor tissues. Fluorescent proteins as the reporter were used to achieve a non-invasive and real-time monitoring of biomarker expression. Fluorescence intensity reduction is observed after testing the biomarker inhibitors, such as YM155 and doxorubicin for survivin, and several natural products for hTERT. This assay offers both molecular and physiological predictions of tissue-specific responses to antitumor agents, thus facilitating early-stage cancer drug discovery.