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Resveratrol Specifically Kills Cancer Cells by a Devastating Increase in the Ca 2+ Coupling Between the Greatly Tethered Endoplasmic Reticulum and Mitochondria

View Article: PubMed Central - PubMed

ABSTRACT

Background/aims: Resveratrol and its derivate piceatannol are known to induce cancer cell-specific cell death. While multiple mechanisms of actions have been described including the inhibition of ATP synthase, changes in mitochondrial membrane potential and ROS levels, the exact mechanisms of cancer specificity of these polyphenols remain unclear. This paper is designed to reveal the molecular basis of the cancer-specific initiation of cell death by resveratrol and piceatannol.

Methods: The two cancer cell lines EA.hy926 and HeLa, and somatic short-term cultured HUVEC were used. Cell viability and caspase 3/7 activity were tested. Mitochondrial, cytosolic and endoplasmic reticulum Ca2+ as well as cytosolic and mitochondrial ATP levels were measured using single cell fluorescence microscopy and respective genetically-encoded sensors. Mitochondria-ER junctions were analyzed applying super-resolution SIM and ImageJ-based image analysis.

Results: Resveratrol and piceatannol selectively trigger death in cancer but not somatic cells. Hence, these polyphenols strongly enhanced mitochondrial Ca2+ uptake in cancer exclusively. Resveratrol and piceatannol predominantly affect mitochondrial but not cytosolic ATP content that yields in a reduced SERCA activity. Decreased SERCA activity and the strongly enriched tethering of the ER and mitochondria in cancer cells result in an enhanced MCU/Letm1-dependent mitochondrial Ca2+ uptake upon intracellular Ca2+ release exclusively in cancer cells. Accordingly, resveratrol/piceatannol-induced cancer cell death could be prevented by siRNA-mediated knock-down of MCU and Letm1.

Conclusions: Because their greatly enriched ER-mitochondria tethering, cancer cells are highly susceptible for resveratrol/piceatannol-induced reduction of SERCA activity to yield mitochondrial Ca2+ overload and subsequent cancer cell death.

No MeSH data available.


Related in: MedlinePlus

Cell viability of EA.hy926 and HUVEC cells was measured via Celltiter-Blue assay according to the standard protocol after 36 h of incubation with resveratrol (Resv; 100 µM), piceatannol (Pice, 100 µM) or oligomycin A (oligo, 10 µM) and calculated as percentage of viable cells normalized to control conditions (A). Caspase activity of EA.hy926 and HUVEC cells, normalized to control conditions as percentage of viable cells, was determined with Caspase 3/7-Glo assay following the standard protocol after 36 h of compound incubation (B).
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Figure 1: Cell viability of EA.hy926 and HUVEC cells was measured via Celltiter-Blue assay according to the standard protocol after 36 h of incubation with resveratrol (Resv; 100 µM), piceatannol (Pice, 100 µM) or oligomycin A (oligo, 10 µM) and calculated as percentage of viable cells normalized to control conditions (A). Caspase activity of EA.hy926 and HUVEC cells, normalized to control conditions as percentage of viable cells, was determined with Caspase 3/7-Glo assay following the standard protocol after 36 h of compound incubation (B).

Mentions: The effects of resveratrol and its derivate piceatannol on cell survival and apoptosis were compared in somatic short-cultured human umbilical vein endothelial cells (HUVEC) with the endothelial/epithelial cancer cell hybrid EA.hy926. Resveratrol and piceatannol had only a small effect on cell viability and caspase 3/7 activity in somatic HUVEC cells (Fig. 1A). In contrast, a 36 h treatment of the cancerous EA.hy926 cells with resveratrol or piceatannol decreased cell viability by more than 60 % and around 70%, respectively (Fig. 1A). Consistently, the activity of apoptotic caspases 3/7 upon treatment with either resveratrol or piceatannol remained unchanged in HUVEC while was increased by more than 7- and 8-fold in EA.hy926 cells (Fig. 1B).


Resveratrol Specifically Kills Cancer Cells by a Devastating Increase in the Ca 2+ Coupling Between the Greatly Tethered Endoplasmic Reticulum and Mitochondria
Cell viability of EA.hy926 and HUVEC cells was measured via Celltiter-Blue assay according to the standard protocol after 36 h of incubation with resveratrol (Resv; 100 µM), piceatannol (Pice, 100 µM) or oligomycin A (oligo, 10 µM) and calculated as percentage of viable cells normalized to control conditions (A). Caspase activity of EA.hy926 and HUVEC cells, normalized to control conditions as percentage of viable cells, was determined with Caspase 3/7-Glo assay following the standard protocol after 36 h of compound incubation (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5382978&req=5

Figure 1: Cell viability of EA.hy926 and HUVEC cells was measured via Celltiter-Blue assay according to the standard protocol after 36 h of incubation with resveratrol (Resv; 100 µM), piceatannol (Pice, 100 µM) or oligomycin A (oligo, 10 µM) and calculated as percentage of viable cells normalized to control conditions (A). Caspase activity of EA.hy926 and HUVEC cells, normalized to control conditions as percentage of viable cells, was determined with Caspase 3/7-Glo assay following the standard protocol after 36 h of compound incubation (B).
Mentions: The effects of resveratrol and its derivate piceatannol on cell survival and apoptosis were compared in somatic short-cultured human umbilical vein endothelial cells (HUVEC) with the endothelial/epithelial cancer cell hybrid EA.hy926. Resveratrol and piceatannol had only a small effect on cell viability and caspase 3/7 activity in somatic HUVEC cells (Fig. 1A). In contrast, a 36 h treatment of the cancerous EA.hy926 cells with resveratrol or piceatannol decreased cell viability by more than 60 % and around 70%, respectively (Fig. 1A). Consistently, the activity of apoptotic caspases 3/7 upon treatment with either resveratrol or piceatannol remained unchanged in HUVEC while was increased by more than 7- and 8-fold in EA.hy926 cells (Fig. 1B).

View Article: PubMed Central - PubMed

ABSTRACT

Background/aims: Resveratrol and its derivate piceatannol are known to induce cancer cell-specific cell death. While multiple mechanisms of actions have been described including the inhibition of ATP synthase, changes in mitochondrial membrane potential and ROS levels, the exact mechanisms of cancer specificity of these polyphenols remain unclear. This paper is designed to reveal the molecular basis of the cancer-specific initiation of cell death by resveratrol and piceatannol.

Methods: The two cancer cell lines EA.hy926 and HeLa, and somatic short-term cultured HUVEC were used. Cell viability and caspase 3/7 activity were tested. Mitochondrial, cytosolic and endoplasmic reticulum Ca2+ as well as cytosolic and mitochondrial ATP levels were measured using single cell fluorescence microscopy and respective genetically-encoded sensors. Mitochondria-ER junctions were analyzed applying super-resolution SIM and ImageJ-based image analysis.

Results: Resveratrol and piceatannol selectively trigger death in cancer but not somatic cells. Hence, these polyphenols strongly enhanced mitochondrial Ca2+ uptake in cancer exclusively. Resveratrol and piceatannol predominantly affect mitochondrial but not cytosolic ATP content that yields in a reduced SERCA activity. Decreased SERCA activity and the strongly enriched tethering of the ER and mitochondria in cancer cells result in an enhanced MCU/Letm1-dependent mitochondrial Ca2+ uptake upon intracellular Ca2+ release exclusively in cancer cells. Accordingly, resveratrol/piceatannol-induced cancer cell death could be prevented by siRNA-mediated knock-down of MCU and Letm1.

Conclusions: Because their greatly enriched ER-mitochondria tethering, cancer cells are highly susceptible for resveratrol/piceatannol-induced reduction of SERCA activity to yield mitochondrial Ca2+ overload and subsequent cancer cell death.

No MeSH data available.


Related in: MedlinePlus