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Hyperthermia-induced apoptosis in Tca8113 cells is inhibited by heat shock protein 27 through blocking phospholipid
1The Affiliated Stomatological Hospital of Kunming Medical College, Kunming, China.
Purpose:
Hyperthermia induces tumour cell apoptosis through the mitochondrial apoptotic pathway; however, the signal transduction mechanism underlying this process still needs to be fully elucidated. Phospholipid scramblase 3 (PLS3), a target of protein kinase C-delta (PKC-delta), resides in mitochondria and plays pivotal roles in regulating apoptotic response. Activated PLS3 facilitates cardiolipin (CL) translocation from the mitochondrial inner membrane to the outer leaflet of the mitochondrial outer membrane and triggers apoptosis.
Materials And Methods:
The tongue squamous cell carcinoma Tca8113 cells were transfected or co-transfected using Lipofectamine 2000 with plasmids pCMV-6xHis-PLS3, pCMV-6xHis-PLS3 (T21A), pHA-PKC-delta, pHA-PKC-delta-KD (K376R), pHA-Hsp27, and empty control plasmid pcDNA3.1. The transfected cells were heated in water bath at 43 degrees C for 20 min, 40 min and 60 min. Assessments of apoptosis and redistribution of mitochondrial cardiolipin were performed by flow cytometry. PLS3, PKC-delta, Hsp27, phosphorylation of PLS3 and PLS3/PKC-delta interaction were detected by western blotting.
Results:
In our study the results show that elevated levels of the wild-type PLS3, but not the PLS3 (T21A) mutant, is able to increase hyperthermia-induced CL translocation and apoptosis. Wild-type PKC-delta facilitates PLS3 phosphorylation, PKC-delta/PLS3 interaction, and CL translocation, which consequently promote apoptosis. In contrast, heat shock protein 27 (Hsp27) blocks PKC-delta-induced PLS3 phosphorylation, suppresses PKC-delta/PLS3 interaction and CL translocation, and inhibits apoptosis.
Conclusions:
Our findings suggest that phosphorylation of PLS3 by PKC-delta is involved in the hyperthermia-induced apoptotic signal transduction pathway in Tca8113 cells, and that Hsp27 blocks this pathway to suppress hyperthermia-induced apoptosis.
Insights
Hyperthermia triggers cancer cell death via mitochondrial pathways. Protein kinase C-delta (PKC-delta) phosphorylates phospholipid scramblase 3 (PLS3), promoting apoptosis, while heat shock protein 27 (Hsp27) inhibits this process.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Hyperthermia induces cancer cell apoptosis via mitochondrial pathways.
- The precise signaling mechanisms require further elucidation.
- Phospholipid scramblase 3 (PLS3) is a key regulator of apoptosis, influenced by protein kinase C-delta (PKC-delta).
Purpose of the Study:
- To investigate the role of PLS3 phosphorylation by PKC-delta in hyperthermia-induced apoptosis.
- To determine the effect of heat shock protein 27 (Hsp27) on this signaling pathway.
Main Methods:
- Tca8113 tongue cancer cells were transfected with plasmids encoding wild-type PLS3, a PLS3 mutant (T21A), wild-type PKC-delta, a kinase-dead PKC-delta mutant, and Hsp27.
- Cells were exposed to hyperthermia (43°C).
- Apoptosis, cardiolipin translocation, protein levels, phosphorylation, and protein interactions were assessed using flow cytometry and western blotting.
Main Results:
- Wild-type PLS3, but not the T21A mutant, enhanced hyperthermia-induced cardiolipin translocation and apoptosis.
- PKC-delta promoted PLS3 phosphorylation, interaction, and cardiolipin translocation, leading to apoptosis.
- Hsp27 inhibited PKC-delta-mediated PLS3 phosphorylation and interaction, thereby suppressing apoptosis.
Conclusions:
- PKC-delta-mediated phosphorylation of PLS3 is crucial for hyperthermia-induced apoptosis signaling in Tca8113 cells.
- Hsp27 acts as an inhibitor of this pathway, suppressing hyperthermia-induced apoptosis.
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