GRIP: A Functional Genomics Strategy to Identify and Validate Metastasis-promoting Gene Networks in Breast Cancer
Abstract: Though survival rates have improved over the past decade, breast cancer continues to be the second leading cause of cancer-related deaths in women. Metastasis causes 90% of all cancer-related deaths and is the reason for the drastic drop in breast cancer survival rate from stage three (79%) to stage four (22%). Conventional methods such as expression array and siRNA screens require considerable effort, and fail to identify specific genes that promote metastasis. To remedy these issues, we have developed a novel genome-wide random insertion of promoter (GRIP) that selects and upregulates a single gene in a random fashion. Therefore, a library of GRIP cells could be screened to identify and select cells that gained metastatic ability. The GRIP-activated genes in selected cells could then easily be identified using PCR techniques. In a proof of concept experiment, To create a breast cancer cell library, GRIP was incorporated into the genome of MCF-7 cells, an estrogen receptor positive with low metastatic potential, and clones were selected. Screening of MCF-7GRIP clones using Boyden-Chamber invasion assay identified two clones with two-fold higher invasiveness than the mock transfected control cells. Moreover, identified clones exhibited migrated faster in wound closing assays, suggesting GRIP is viable functional genomic strategy to identify metastasis-promoting genes. By identifying the individual gene upregulated in each of these clones, we can provide a therapeutic target for breast cancer metastasis.
Monekah Ammouri*, Zara Barati, and Venugopalan Cheriyath, PhD
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