c-MYC’s role on methylation of the GATA2 gene in non-small cell lung carcinoma Jonathan Hajduk

Cancer Caused through random mutation of the Genome Improper cell division, ultraviolet light, natural and man-made mutagenic chemicals and reactive oxygen species generated by ionizing radiation Cancer prevention methods Cell inviability after mutation Cytotoxic T cells Natural killer cells Lung cancer 225,000 new cases a year in the U.S.

GATA2 GATA family is a group of transcription factors regulate gene expression of hematopoietic cells this includes: platelets, red blood cells, mast cells, basophils, neutrophils, eosinophils, macrophages, B cells, and T cells GATA2 is expressed in Natural Killer cells Innate immune system looking for improperly regulated MHC-1 molecules

GATA2

Key factors Fhit gene Methylation RAS pathway Wnt Pathway Methylated by c-Myc Methylation RAS pathway cell proliferation, cell differentiation, cell adhesion, apoptosis, and cell migration Wnt Pathway regulation of gene transcription, calcium levels

c-MYC’s role in the cell cell proliferation, metabolism, differentiation, and apoptosis Known effects on miR-29b and Fhit 85% of NSCLC involves this pathway Affects genes through mRNA

Methods NCI-H522 cell line c-MYC expression “knocked down” using siRNA Cells split weekly to prevent overgrowth c-MYC expression “knocked down” using siRNA DNA isolated from the cells using Promega Protocol Bisulfite sequencing on isolated DNA PCR reaction and DNA clean-up

Where are we? siRNA knock down complete DNA isolation complete Bisulfite treatment complete Next Steps: PCR analysis to be completed.

Further Work Repetition of the experiment Evidence of pathway factors on GATA2 How does a lack of Ras or Wnt impact GATA2? How does Ras, Wnt, and c-MYC impact GATA2 in vivo? Other potential means of mutating or preventing GATA2

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