Loss of protein kinase Cδ/HuR interaction is necessary to doxorubicin resistance in breast cancer cell lines.
The protein kinase Cδ (PKCδ) interacts with and phosphorylates HuR, dictating its functionality. We show here that the genotoxic stimulus induced by doxorubicin triggers PKCδ interaction with HuR and leads to HuR phosphorylation on serines 221 and 318 and cytoplasmic translocation. This series of events is crucial to elicit the death pathway triggered by doxorubicin and is necessary to promote HuR function in post-transcriptional regulation of gene expression, because genetic ablation of PKCδ caused the inability of HuR to bind its target mRNAs, topoisomerase IIα (TOP2A) included. In in vitro select doxorubicin-resistant human breast cancer cell lines upregulating the multidrug resistance marker ABCG2, PKCδ, and HuR proteins were coordinately downregulated together with the doxorubicin target TOP2A protein whose mRNA was HuR-regulated. Therefore, we show here that PKCδ, HuR, and TOP2A constitute a network mediating doxorubicin efficacy in breast cancer cells. The importance of these molecular events in cancer therapy is suggested by their being profoundly suppressed in cells selected for doxorubicin resistance.
Laboratory of Genomic Screening (E.L., I.C., A.P.) and Laboratory of Translational Genomics (P.G., A.Q.), Centre for Integrative Biology, University of Trento, Italy; and Pharmacology Laboratory, Health Sciences Department, University of Milan, Italy (E.L., S.C.).
Elisa
E
Latorre
Ilaria
I
Castiglioni
Pamela
P
Gatto
Stephana
S
Carelli
Alessandro
A
Quattrone
Alessandro
A
Provenzani
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001943:Breast Neoplasms / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004317:Doxorubicin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D051959:ELAV Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D051745:Protein Kinase C-delta / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics
D000903:Antibiotics, Antineoplastic; D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D051959:ELAV Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D034741:RNA, Small Interfering; D004317:Doxorubicin; D051745:Protein Kinase C-delta; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human