DMSO Treatment of K562 Cells to Induce Erythrocytic Differentiation Increases γ-Globin Expression and Inhibits Apoptosis

Abstract

The overall objective of this study was to test if a 210mM concentration of DMSO can induce erythrocytic differentiation in K562 leukemia cells. Verification of erythrocytic differentiation was attempted using multiple experiments that compare characteristic differences found in induced versus uninduced K562 cells, including a growth curve measuring cell growth, benzidine staining to measure hemoglobin expression, an agarose gel electrophoresis to detect levels of gamma-globin RNA expression, a western blot to detect levels of Hbγ and Hbα protein expression, and a caspase-3 colorimetric apoptosis assay to detect levels of apoptosis. The growth curve and agarose gel did not indicate significant differences in DMSO-treated or untreated K562 cells in terms of growth or γ-globin RNA expression. However, the western blot and benzidine staining experiments did indicate greater expression of Hbγ + Hbα and hemoglobin, respectively, in the DMSO-treated K562 cells versus the untreated K562 cells. Unexpectedly though, the capase-3 assay showed a greater colorimetric response from untreated K562 cells versus DMSO-treated K562 cells. The results from these experiments provided evidence at the protein level that a 210 mM DMSO treatment of K562 cells was effective in inducing erythrocytic differentiation, while also inhibiting apoptosis of those treated cells.

Faculty Mentors: Nina Bernstein and Kimberley Harcombe

Department: Biological Science