Adam Al Douri/ Devon Beauchamp

The Effects of TPA and Hemin on the K562 Cell Line
Adam H. Al Douri, Devon Beauchamp
Major(s): Biology, Psychology
Faculty Sponsor: Dr. Tara Allen
The K562 cell line was derived from a fifty-three year old Caucasian female
patient with chronic mylogenous leukemia. This study will examine the ability of the
K562 cell line to differentiate into megakaryocytic cells, when treated with a signaling
Hemin, as well as erythrocytes when treated with Phorbol-12-myristate-13-acetate
(TPA). The cells were grown in RPMI media in an environment that was composed of
5% CO 2 at a temperature of 37 o C. Differentiation of K562 cells was carried out through
treatment of the cells with 50 µM of Hemin, 9.7 nM of TPA, or a combination of 9.7 nM
TPA and a 6.0 µM of Hemin for a period of 24 hours. The incubation time as well as the
concentrations of the chemicals used was set using past experimental data and primary
literature findings. Observations were made at the conclusion of the experiment by
extracting 100 µL of solution from each treatment to undergo analysis via
hemocytometer to estimate the amount of cell proliferation. Further analysis was
conducted using flow cytometery with Propidium iodide (PI) analysis. PI was done to
indicate which phase of the cell cycle (G 1 , G 2 /M, and sub G 1 ) the observed cells are in.
We observed a large amount of cells residing in the sub G 1 phase in all treated flasks
compared to those observed in our control. This presence in the sub G 1 phase is
indicative of cell apoptosis. This cell death could be due to a combination of the toxic
nature of both chemicals and the incubation duration. As for our flow analysis we
expected to see a reduction in size when cells were differentiated into erythrocytes and

an increase in cell size when differentiation into megakaryocytic cells had occurred. To
validate our hypothesis the results of our flow were depicted using two different graphs:
FSC and SSC. The FSC was used to indicate cell size. Here we saw a shift away from
the control group in all tested lines. The SSC was used to indicate cell complexity. Here
we saw a decrease in cell complexity when treated with TPA and a slight increase when
treated with Hemin from the complexity of our control group. These variations from the
control group indicate possible cell differentiation into erythrocytes as well as
megakaryocytic cells. Overall the possible differentiation of the K562 cell line could
open up new avenues for cancer research.