Dynamic Control of Mitochondrial Ca2+ Levels as a Survival Strategy of Cancer Cells

Abstract

Cancer cells have increased energy requirements due to their enhanced proliferation activity. This energy demand is, among others, met by mitochondrial ATP production. Since the second messenger Ca²⁺ maintains the activity of Krebs cycle dehydrogenases that fuel mitochondrial respiration, proper mitochondrial Ca²⁺ uptake is crucial for a cancer cell survival. However, a mitochondrial Ca²⁺ overload induces mitochondrial dysfunction and, ultimately, apoptotic cell death. Because of the vital importance of balancing mitochondrial Ca²⁺ levels, a highly sophisticated machinery of multiple proteins manages mitochondrial Ca²⁺ homeostasis. Notably, mitochondria sequester Ca²⁺ preferentially at the interaction sites between mitochondria and the endoplasmic reticulum (ER), the largest internal Ca²⁺ store, thus, pointing to mitochondrial-associated membranes (MAMs) as crucial hubs between cancer prosperity and cell death. To investigate potential regulatory mechanisms of the mitochondrial Ca²⁺ uptake routes in cancer cells, we modulated mitochondria-ER tethering and the expression of UCP2 and analyzed mitochondrial Ca²⁺ homeostasis under the various conditions. Hence, the expression of contributors to mitochondrial Ca²⁺ regulation machinery was quantified by qRT-PCR. We further used data from The Cancer Genome Atlas (TCGA) to correlate these in vitro findings with expression patterns in human breast invasive cancer and human prostate adenocarcinoma. ER-mitochondrial linkage was found to support a mitochondrial Ca²⁺ uptake route dependent on uncoupling protein 2 (UCP2) in cancer cells. Notably, combined overexpression of Rab32, a protein kinase A-anchoring protein fostering the ER-mitochondrial tethering, and UCP2 caused a significant drop in cancer cells' viability. Artificially enhanced ER-mitochondrial tethering further initiated a sudden decline in the expression of UCP2, probably as an adaptive response to avoid mitochondrial Ca²⁺ overload. Besides, TCGA analysis revealed an inverse expression correlation between proteins stabilizing mitochondrial-ER linkage and UCP2 in tissues of human breast invasive cancer and prostate adenocarcinoma. Based on these results, we assume that cancer cells successfully manage mitochondrial Ca²⁺ uptake to stimulate Ca²⁺-dependent mitochondrial metabolism while avoiding Ca²⁺-triggered cell death by fine-tuning ER-mitochondrial tethering and the expression of UCP2 in an inversed manner. Disruption of this equilibrium yields cancer cell death and may serve as a treatment strategy to specifically kill cancer cells.

Keywords: ER stress; cancer cells; mitochondrial Ca2+ homeostasis; mitochondrial-ER interaction; uncoupling protein 2.