The Effects of Triptolide on Cellular Metabolism in Pancreatic Cancer and Fibroblast Cells

Morgan Frank
Pancreatic Cancer Research Lab
Abstract
Background: Pancreatic cancer is the third leading cause of adult cancer death in the United States
and each year approximately 41,800 patients die from the disease. Late diagnosis, limited
treatment options, and low response rates to current standard of care make PC extremely lethal
with devastatingly low survival rates (>94% of patients die within 5 years of diagnosis). One of
the primary reasons for the dismal prognosis is that the tumor microenvironment (TME) in PC is
incredibly dense, fibrotic, and hypo-vascular making it impenetrable to chemotherapeutics.
Triptolide (Minnelide), a compound with anti-tumor activity, is in Phase I clinical trials to treat
various solid tumors and is showing promising activity in several cancer types including PC.
Objective: Targeting the mitochondrial and glycolytic cellular energy pathways is an effective way
to inhibit tumor cell growth. Therefore, this project aimed at elucidating the effect of triptolide on
the cellular energetics of PC (MIAPaca-2) and fibroblast (PS1) cells, which predominate the TME.
Methods: The Seahorse XFe96 Analyzer was employed to probe cellular energy pathways
including glycolysis and oxidative phosphorylation in MIAPaca-2 and PS1. The XF Cell Mito
Stress Test (MST) measures oxygen consumption rate (OCR) to evaluate mitochondrial function
and the XF Cell Glycolysis Stress Test (GST) detects changes in extracellular acidification rate
(ECAR) to evaluate glycolytic function. Dose response and kinetic time course experiments were
performed on triptolide treated MIAPaca-2 and PS1 cells. This was done to determine the
minimal dose that elicits a significantly different response in oxidative phosphorylation and
glycolysis as well as its time course of action when compared to control. Cells pretreated with

different concentrations of triptolide (0.03uM to 3uM) for varying times up to six hours were
subjected to the GST or MST.
Results: Our data suggests that triptolide reduces mitochondrial respiration and glycolysis in a
dose dependent manner in MIAPaCa-2 and PS1 cells within six hours of treatment. Under the
tested conditions, 0.3uM triptolide significantly reduced OCR and ECAR in both cell lines. This
response is evident in key parameters of mitochondrial function (ATP production, maximal
respiration, and spare respiratory capacity) and glycolytic function (glycolytic capacity and
glycolytic reserve).
Conclusions and Future Directions: Triptolide adversely affects glycolytic and mitochondrial
function in pancreatic cancer cells and fibroblasts. Future research should include similar
experiments using other pancreatic cancer cell lines and correlate the effects on mitochondrial and
glycolytic function to anti-tumor activities to provide a better understanding of the drugs
mechanism of action. Probing cellular energy pathways in cancer cells treated with triptolide in
combination with currently used treatment options and/or other new agents may lead to a better
alternative to treat PC.