(190ab) Engineering Cancer Cells for Cancer Research

The use of engineered cancer cells can greatly expedite and facilitate cancer research. Unfortunately, the arsenal of commercially available recombinant cancer cell lines is limited, and effective transfection of cancer cell lines is challenging, as transfection protocols often need customizing for each cell line.

Here, we present a set of simple lipotransfection protocols for genetic engineering of three different commercial cancer cell lines widely used in cancer research. We have used this technique for the successful transfection of vectors of different sizes into DU145 (prostate), PC3 (prostate), and MCF7 (breast) cancer cells. For example, we produced pools of fluorescent cancer cells by lipotransfection of constructs encoding the expression of green fluorescent protein (GFP), red fluorescent protein (RFP), and blue fluorescent protein (BFP). These pools maintained expression for at least 5 months, allowing long-term culture experiments and facilitating fluorescence microscopy visualization without using cell dyes. We also demonstrated the transfection of DU145 cancer cells with green and red fluorescent proteins (GFP and RFP) as reporters encoded by the GAV and Z-NEP vectors. These vectors enable the “on-demand” emission of fluorescence upon exposure to blue light (λ=450 nm).

We envision multiple applications of these cancer engineer cells in biomedical settings. For example, the co-culture of healthy and cancerous cells expressing different fluorescent proteins might be useful research in cancer-on-chip studies. The on-demand expression of recombinant proteins in cancer cells can be a powerful therapeutic tool (i.e., controlled release of anti-cancer proteins mediated through light or other stimuli).