Exploration of Epigenetic Modifications in Taxus Cell Culture using Global Demethylating Agents

Whittles, Emily A.; Gamache, Michaela F.

Student Work

Exploration of Epigenetic Modifications in Taxus Cell Culture using Global Demethylating Agents

Public

Paclitaxel is a widely used chemotherapeutic drug derived from yew trees (Taxus species). Meeting world demand for paclitaxel has historically been difficult, as it is produced in very low concentrations within the bark of the yew tree. Plant cell culture is one sustainable method for paclitaxel production, but as cell lines get older and are continuously subcultured, production decreases. This decreased production is correlated with increased DNA methylation, which is an epigenetic mechanism that downregulates gene expression. This MQP sought to find a global demethylating agent that could reverse this to maintain high paclitaxel yields, even in cultures that have been subcultured for many years. Two cell lines were treated with either 5-azacytidine or zebularine (two global demethylating agents with different mechanisms of action) at either 100μM or 200μM. Half of the flasks were elicited with methyl jasmonate, and half were mock-elicited. Levels of paclitaxel and its precursors, as well as flavonoids and phenolics, were sampled and measured weekly over the course of the 21-day long experiment. Treatment, while visually looking to positively correlate with paclitaxel production, was found to have no statistically significant effect on paclitaxel production. There was also no effect on the production of paclitaxel precursors 10-deacetylbaccatin and baccatin. It was further found that there was no difference between 5-azacytidine and zebularine or between the different concentrations. Notably, there was also no difference between the elicited and mock-elicited flasks. In addition, treatment had no discernable effect on flavonoid and phenolic production.

This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

Creator

Subject

Publisher