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

2. 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.

3. 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

4. GATA2

5. 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

6. 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

7. 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

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

9. 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

10. References
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