Oxidative Stress-Induced Unscheduled CDK1-Cyclin B1 Activity Impairs ER-Mitochondria-Mediated Bioenergetic Metabolism

Jan-Gowth Chang; Ni Tien; Yi-Chih Chang; Meng-Liang Lin; Shih-Shun Chen

doi:10.3390/cells10061280

Oxidative Stress-Induced Unscheduled CDK1-Cyclin B1 Activity Impairs ER-Mitochondria-Mediated Bioenergetic Metabolism

Cells. 2021 May 21;10(6):1280. doi: 10.3390/cells10061280.

Authors

Jan-Gowth Chang¹, Ni Tien¹, Yi-Chih Chang², Meng-Liang Lin³, Shih-Shun Chen^{2

4}

Affiliations

¹ Department of Laboratory Medicine, China Medical University Hospital, Taichung 404394, Taiwan.
² Department of Medical Laboratory Science and Biotechnology, College of Medical and Health Science, Asia University, Taichung 41354, Taiwan.
³ Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404394, Taiwan.
⁴ Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung 406053, Taiwan.

Abstract

Targeting the activities of endoplasmic reticulum (ER)-mitochondrial-dependent metabolic reprogramming is considered one of the most promising strategies for cancer treatment. Here, we present biochemical subcellular fractionation, coimmunoprecipitation, gene manipulation, and pharmacologic evidence that induction of mitochondria-localized phospho (p)-cyclin dependent kinase 1 (CDK1) (Thr 161)-cyclin B1 complexes by apigenin in nasopharyngeal carcinoma (NPC) cells impairs the ER-mitochondrial bioenergetics and redox regulation of calcium (Ca⁺⁺) homeostasis through suppressing the B cell lymphoma 2 (BCL-2)/BCL-2/B-cell lymphoma-extra large (BCL-x_L)-modulated anti-apoptotic and metabolic functions. Using a specific inducer, inhibitor, or short hairpin RNA for acid sphingomyelinase (ASM) demonstrated that enhanced lipid raft-associated ASM activity confers alteration of the lipid composition of lipid raft membranes, which leads to perturbation of protein trafficking, and induces formation of p110α free p85α-unphosphorylated phosphatase and tensin homolog deleted from chromosome 10 complexes in the lipid raft membranes, causing disruption of phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt)-GTP-ras-related C3 botulinum toxin substrate 1 (Rac1)-mediated signaling, thus triggering the p-CDK1 (Thr 161))-cyclin B1-mediated BCL-2 (Thr 69/Ser 87)/BCL-x_L (Ser 62) phosphorylation and accompanying impairment of ER-mitochondria-regulated bioenergetic, redox, and Ca⁺⁺ homeostasis. Inhibition of apigenin-induced reactive oxygen species (ROS) generation by a ROS scavenger N-acetyl-L-cysteine blocked the lipid raft membrane localization and activation of ASM and formation of ceramide-enriched lipid raft membranes, returned PI3K-Akt-GTP-Rac1-modulated CDK1-cyclin B1 activity, and subsequently restored the BCL-2/BCL-x_L-regulated ER-mitochondrial bioenergetic activity. Thus, this study reveals a novel molecular mechanism of the pro-apoptotic activity of ASM controlled by oxidative stress to modulate the ER-mitochondrial bioenergetic metabolism, as well as suggests the disruption of CDK1-cyclin B1-mediated BCL-2/BCL-x_L oncogenic activity by triggering oxidative stress-ASM-induced PI3K-Akt-GTP-Rac1 inactivation as a therapeutic approach for NPC.

Keywords: ASM; BCL-2/BCL-xL; ER–mitochondria; apigenin; cyclin B1–CDK1; lipid raft; oxidative stress.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
CDC2 Protein Kinase / physiology*
Cell Line, Tumor
Cyclin B1 / physiology*
Endoplasmic Reticulum / metabolism*
Endoplasmic Reticulum / pathology
Female
Humans
Male
Middle Aged
Mitochondria* / metabolism
Mitochondria* / pathology
Nasopharyngeal Carcinoma / metabolism*
Oxidative Stress

Substances

CCNB1 protein, human
Cyclin B1
CDC2 Protein Kinase
CDK1 protein, human