Abstract
The Lin et al. (2012), published in Cell in 2012. The experiments that will be replicated are those reported in Figures 3E and 3F. In these experiments, elevated levels of c-Myc in the P493-6 cell model of Burkitt's lymphoma results in an increase of the total level of RNA using UV/VIS spectrophotometry (Figure 3E; Lin et al., 2012) and on the mRNA levels/cell for a large set of genes using digital gene expression technology (Figure 3F; Lin et al., 2012). The Reproducibility Project: Cancer Biology is a collaboration between the eLife.
Highlights
The proto-oncogene MYC is frequently amplified in human cancers and encodes the transcription factor c-Myc, which is associated with a variety of cellular processes such as cell growth and proliferation (Dang, 2013)
While overexpressed c-Myc is known to contribute to tumorigenesis, an understanding of how this process occurs is complicated by a number of issues, including the large number of binding sites and the diversity between systems
P493-6 cells will be cultured with two separate lots of serum that will be maintained throughout all the experiments to assess if there is variability between different batches of serum
Summary
The proto-oncogene MYC is frequently amplified in human cancers and encodes the transcription factor c-Myc, which is associated with a variety of cellular processes such as cell growth and proliferation (Dang, 2013). An increase in total cellular RNA content was observed in 3T9 fibroblasts following serum stimulation, which was not observed when c-Myc was deleted in these cells (Sabò et al, 2014) This experiment is replicated in Protocol 2. Two papers were published that used primarily RNAseq and ChIP-seq to focus on assessing if the transcriptional effects of c-Myc are direct or indirect (Alderton, 2014) These studies found that RNA amplification and promoter/enhancer invasion by c-Myc were separable events in 3T9 fibroblasts and U2OS cells, suggesting that c-Myc regulates a distinct subset of genes, which indirectly lead to RNA amplification (Sabò et al, 2014; Walz et al, 2014)
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