Determining the Methylation Status of Chordoma Cell Lines Using Methylation Specific PCR (MSP)
Chordomas are slow growing tumors associated with poor clinical outcomes due to their difficulty to treat. One gene that may play a role in chordoma development is the LYST gene, which is frequently affected by loss-of-function mutations. Preliminary work by the Venteicher lab has indicated that this gene may be methylated in certain chordoma subtypes, an epigenetic change that would have a similar effect as a loss-of-function mutation. To verify this hypothesis, methylation-specific PCR (MSP) was utilized to analyze the methylation status of the following chordoma cell lines: CH1, UM-Chor1, CH7, JCH7, CH22, U-CH2, and MUG-CC1. Discovery of a chordoma cell line containing methylation of the LYST gene would allow further testing of effect on tumor proliferation. If physicians can differentiate between more and less severe subtypes of chordoma using simple methylation analysis, then they may be able to avoid harsher treatments (e.g. radiotherapy) for relatively milder cases, improving patient outcomes. None of the cell lines were methylated in the region of interest; future research should focus on testing more cell lines with primers targeting different regions. Additionally, quantitative measures could be used to detect if there is some degree of methylation, rather than simply viewing the binary output that MSP provides.