What type of study is this?

This is a In Vitro Study study (also classified as: Animal Study).

September 10, 2025Open Access

Figure 5 from PPARδ Orchestrates a Prometastatic Metabolic Response to Microenvironmental Cues in Pancreatic Cancer

Key Points

Targeting PPARδ impacts both glycolysis and OXPHOS, linking metabolic regulation to metastasis.
Evidence from assays shows that inhibition of PPARδ alters cell invasion patterns in PDAC cells.
The methodology involved in vitro and in vivo approaches using pancreatic cancer cell lines and mouse models.
Findings support further investigation into metabolic therapies targeting PPARδ in pancreatic cancer.

Abstract

PPARδ controls the balance between OXPHOS and glycolysis, linked to EMT and metastasis. A,In vitro invasion in PDAC-215, 253, and 354 cells stably transduced with inducible lentiviral vectors expressing either a nontargeting short hairpin RNA (NT shRNA) or three different shRNAs against PPARD (sh#1, sh#2, and sh#3). Transduced cells were pretreated with doxycycline for 24 hours, then incubated with MCM, etomoxir (Eto), or L-165 for 48 hours, and finally plated in modified Boyden chambers for 16 hours (n = 7). B, Top, ZsGreen expression by RT-qPCR in liver homogenates from an in vivo metastasis assay of PDAC-354 cells stably expressing either the NT or the sh#1 against PPARD. Cells were pretreated with doxycycline and/or 20 μmol/L etomoxir for 48 hours. After intrasplenic implantation, mice were treated with oral doxycycline (2 mg/mL; drinking water) and etomoxir (15 mg/kg, i.p. daily) for 7 days, when splenectomies were performed. Bottom, numbers indicate the percentage and total number of micrometastases in each experimental group. C, PDAC-215, 253, and 354 transduced cells as in A were pretreated with doxycycline for 24 hours, then incubated with MCM, etomoxir, or L-165, and then tested for ATP-linked respiration (top) and glycolytic capacity (bottom) after additional 24 hours (n = 8). D, Mitochondrial stress test (top row) and glycolysis test (bottom row) following treatment with control (Ctrl) or the PPARδ agonists L-165 or GW0742. Left column, representative OCR and extracellular acidification rate (ECAR) profiles for PDAC-253. Right column, pooled data for PDAC-215, 253, and 354 cells (n = 6–9). Glyco, glycolytic; Max res, maximum respiration; O, ATP synthase inhibitor oligomycin; F, mitochondrial OXPHOS uncoupler FCCP carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone; A+R, complex III inhibitor antimycin A + electron transport change inhibitor rotenone. G, glucose; 2DG, glycolysis inhibitor 2-deoxy-glucose. E, ATP-linked respiration (left) and maximal respiration (right) for control vs. GW0742-treated cells following treatment with or without palmitate-BSA (FAO assay). PDAC-354 cells were treated with 10 μmol/L GW0742 for 48 hours prior to the assay (n = 5). In A, C, and D, the bars represent pooled data from PDAC-215, 253, and 354, showing individual data points corresponding to each PDX. All data are represented as the mean ± SEM. *, P P P P

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