Integrated proteomics and metabolomics reveal phytosesquiterpene lactones DET and DETD-35 inhibit triple-negative breast cancer cell activity by depleting ATP synthesis and reprogramming primary metabolism
Description
Phytosesquiterpene lactones DET and DETD-35 are reported to induce oxidative stress towards inhibiting the activities of triple-negative breast cancer (TNBC) cells. However, how DET and DETD-35 affect mitochondrial proteome and associated oxidative phosphorylation in TNBC cells is unclear. In this study, the mechanisms through which DET and DETD-35 inhibit MDA-MB-231 TNBC cells were dissected. DET and DETD-35 promoted mitochondrial superoxide production by upregulating expression of SOD1 and SOD2, and induced permeability transition pore opening, and attenuated intracellular ATP levels. Neither compound interfered with mitochondrial respiration/bioenergetics in normal mammary MCF-10A cells. Comparative mitochondria proteome and bioinformatic analyses showed significant deregulation of proteins related to the oxidative phosphorylation, depolarization of mitochondria, and apoptosis signaling in DET- or DETD-35-treated TNBC cells, and primary metabolomics revealed that both compounds deregulated metabolites dynamics and the corresponding metabolic pathways in TNBC cells. Knockdown of the PRKCA gene/protein involved in inducing mitochondrial toxicity in TNBC cells reversed cytotoxicity and apoptosis and decreased the levels of several metabolites induced by DET or DETD-35 in the cancer cells. Integrated Pearson’s correlation and IPA network analyses of differentially expressed proteins and metabolites revealed the networks of ATP synthesis, energy homeostasis, and mitochondrial activities (respiration, depolarization, and transmembrane potential) highly correlated to the compound effects. Notable, DET/DETD-35 inhibited mitochondrial ATPase activity, and molecular modeling further predicted the binding sites of either compound with ATP synthase at the subunits α/βand c/a interfaces. In summary, the mechanisms through which these phytosesquiterpene lactones disrupt mitochondrial biogenetics to inhibit TNBC cell activities were delineated.