The detection of differences between normal and

cancerous stem cells through analysis of

morphology, gene expression, and effects of DCA

Justina Cho, Jimmy Liu

Advisers: Ms. Marie Olszewski,
Dr. Morris Kletzel
Acknowledgements
We would like to thank Ms. Marie Olszewski, Dr.
Morris Kletzel, Wei Huang, and Dr. Judith A.
Scheppler for their continual support and
guidance.
Introduction
• Dichloroacetate as a promising cancer therapy
• Mitochondrial action
• Versatility
• Non-toxic
• Previous studies
Methods
• Culturing cells
• Examining gene expression
▫ RT-PCR
• Testing cell viability
▫ Trypan-Blue
• Applying dichloroacetate (DCA)
Results

Figure 1. Bar graph of the cell concentration as time

increases. K562 was cultured and the cell
concentration of the controls (without DCA) were
recorded at various times in a 14 day period.
Results

Figure 2. Protocol a: The effects of increasing DCA

concentration on the cell concentration of K562. Cell
concentration (K/uL) of K562 cell cultures with no DCA
and cultures with 2.5 mM, 5mM, 10mM, and 20mM
were observed after 168hrs after the addition of DCA.
Results

Figure 3. Protocol b: K562 cell concentrations on day

14 of the culture and 96 hours after adding DCA to
certain cultures. 10mM and 20mM DCA concentrations
were tested and compared to the day 14 control
Results

Figure 4. Protocol c: BM2, a normal stem cell sample,

was treated with various concentrations of DCA. The
resulting cell concentrations after 48 hrs of immersion
in DCA were compared to the control, which had been
cultured for a total of 14 days.
Results

Figure 5. WT1 expression in K562 controls, 10mM, 20

mM, and BM2. WT1 expression is relatively low for
BM2, and K562 cells in 20mM DCA concentrations.
Results

Figure 6. WT1 expression in K562 controls, 10mM and

20mM DCA treated K562 samples, and BM2. The
resulting data was examined through the LightCycler
Data Analysis.
Discussion
• Control group K562 proliferates normally
• Protocol A: increased DCA concentration
correlated with decreased cell concentration
▫ Decreased cell proliferation
• Protocol B: Confirming efficacy of 10mM and
20mM concentrations of DCA after 96 hours
Discussion
• DCA tested on non-cancerous stem cells
▫ No effects detected
• DCA inhibits cell proliferation by restoring
apoptosis (Bonnett et al.)
• WT1 gene controls cell proliferation
• WT1 expression in samples
Conclusion
• Increased DCA concentration correlated with
decreased K562 proliferation
• WT1 gene inhibited in 20mM DCA-treated group
• DCA does not affect non-cancerous cells
• Viable cancer treatment
What the future has in store…
• Further DCA tests to solidify data
• Test cancerous stem cells
• Test DCA with samples of healthy and cancerous
cells combined
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Jørgensen, H., Holyoake, T. (2007). Characterization of cancer stem
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