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A Screen of shRNAs Targeting Tumor Suppressor Genes to Identify Factors Involved in Paclitaxel Sensitivity

atients with tumors containing high levels of some genes, such as p53, would be expected to respond less well to paclitaxel, perhaps falling into the group of 80% non-responders, while patients with low levels of other genes, such as VHL, would be expected to respond better to this drug (perhaps falling into the 20% clinical responder population).

An advantage to the use of shRNA is that it allows an investigator to rapidly validate results in multiple model systems. We are encouraged by our findings in this one non-small cell lung carcinoma line and want to attempt to validate the results in additional lines. We are similarly interested in determining whether these genes play a role only in NSCLC, or if they also affect response rates in cell lines derived from other tumor types. We can do this fairly simply with lentiviral delivery of shRNA with little, or no, additional optimization of conditions necessary. Unlike the more traditional siRNA approach, we do not need to optimize transfection or reaction conditions for each new cell line transduction is usually as simple as growing cells and adding lentivirus to the media. Also unlike siRNA, shRNA allows one to have a long-term down-regulation of a gene. This will facilitate our moving from an in vitro setting to an in vivo experiment. After confirmation of our results in mouse xenograft models, we hope to examine the clinical setting to see if patients who have been treated with paclitaxel in the past have a gene expression profile correlating with our findings. If so, these elucidated genes will have exciting possibilities, either as targets or biomarkers for paclitaxel sensitivity.

For more information on the MISSION TRC shRNA collections, visit sigma-aldrich.com/rnai.

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