- •Chemoresistance is the reason for the limited efficacy of breast cancer therapeutics.
- •Receptor activator of nuclear factor-kappa B ligand (RANKL) mediates chemoresistance in breast cancer cells through autophagy induction.
- •RANKL induces autophagy by activating the Janus protein tyrosine kinase-signal transducer and activator of transcription 3 (JAK-STAT3) signaling pathway.
- •Targeting RANKL/receptor activator of nuclear factor-kappa B (RANK) may be a potential strategy to overcome chemoresistance in breast cancer patients.
This study was to investigate the functional role and mechanism of receptor activator of nuclear factor-kappa B ligand (RANKL) associated autophagy and chemoresistance in breast cancer.
Materials and Methods
Cell Counting Kit-8 (CCK-8) assay was used to detect the cell viability. Real-time polymerase chain reaction (PCR) was used for determining the relative mRNA levels of key genes and protein expression was assessed by Western blotting. Immunofluorescence was performed to evaluate the changes in the autophagy flux. Short hairpin (shRNA) was used to knockdown the expression of the target genes in breast cancer cells. Based on The Cancer Genome Atlas (TCGA) database, we explored the expression of receptor activator of nuclear factor-kappa B (RANK), autophagy and signal transducer and activator of transcription 3 (STAT3) signaling associated genes and analyzed their correlation with the prognosis of breast cancer patients.
The findings showed that receptor activator of nuclear factor-kappa B ligand (RANKL), the ligand of RANK, could effectively enhance the chemoresistance potential of breast cancer cells. Our results showed that RANKL induced autophagy and enhanced the expression of autophagy associated genes in breast cancer cells. The knockdown of RANK suppressed RANKL mediated autophagy induction in these cells. Furthermore, the inhibition of autophagy suppressed RANKL mediated chemoresistance in breast cancer cells. We found STAT3 signaling pathway was involved in RANKL-induced autophagy. Analysis of the expression of RANK, and autophagy and STAT3 signaling associated genes in breast cancer tissues showed that the expression of autophagy and STAT3 signaling associated genes was correlated with the prognosis of breast cancer patients.
The present study suggests that the RANKL/RANK axis may potentially mediate chemoresistance in breast cancer cells by inducing autophagy through the STAT3 signaling pathway.
Abbreviations:RANKL (receptor activator of nuclear factor-kappa B ligand), RANK (receptor activator of nuclear factor-kappa B), STAT3 (signal transducer and activator of transcription 3), LC3-II (Microtubule-associated protein 1 light chain 3 isoform II), ATG5 (autophagy-related 5), TNBC (triple-negative breast cancer), DMEM (Dulbecco's modified Eagle's medium), FBS (fetal bovine serum), GADPH (glyceraldehyde-3-phosphate dehydrogenase), PBS (phosphate-buffered saline), ADM (Adriamycin), 5-FU (5-fluorouracil), CQ (chloroquine), CCK8 (cell counting kit-8), OD (optical density), qRT-PCR (Quantitative real-time polymerase chain reaction), GFP (green fluorescent protein), ECL (enhanced chemiluminescence), shRNA (short hairpin RNA), TCGA (The Cancer Genome Atlas), MAP1LC3 (microtubule-associated protein 1 light chain 3), CSCs (Cancer stem cells), JAK (Janus protein tyrosine kinase), SOCS (suppressor of cytokine signaling), PTPN (protein tyrosine phosphatase non-receptor type)
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- Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CA Cancer J Clin. 2021; 71: 209-249
- Breast cancer stem-like cells in drug resistance: a review of mechanisms and novel therapeutic strategies to overcome drug resistance.Front Oncol. 2022; 12856974
- Recent advances in understanding tumor stroma-mediated chemoresistance in breast cancer.Mol Cancer. 2019; 18: 67
- Exosomes and breast cancer drug resistance.Cell Death Dis. 2020; 11: 987
- Targeting autophagy in cancer.Nat Rev Cancer. 2017; 17: 528-542
- The prospects of therapeutic potential and drug development targeting autophagy in cancer.Adv Exp Med Biol. 2020; 1207: 663-679
- Autophagy inducers in cancer.Biochem Pharmacol,. 2018; 153: 51-61
- RANK-ligand (RANKL) expression in young breast cancer patients and during pregnancy.Breast Cancer Res. 2015; 17: 24
- Poor prognosis of patients with triple-negative breast cancer can be stratified by RANK and RANKL dual expression.Breast Cancer Res Treat. 2017; 164: 57-67
- RANKL/RANK pathway abrogates cetuximab sensitivity in gastric cancer cells via activation of EGFR and c-Src.Onco Targets Ther. 2017; 10: 73-83
- RANKL-induced c-Src activation contributes to conventional anti-cancer drug resistance and dasatinib overcomes this resistance in RANK-expressing multiple myeloma cells.Clin Exp Med. 2019; 19: 133-141
- RANK-RANKL interactions are involved in cell adhesion-mediated drug resistance in multiple myeloma cell lines.Tumour Biol. 2016; 37: 9099-9110
- RANK ligand modulation of autophagy in oral squamous cell carcinoma tumor cells.J Cell Biochem. 2016; 117: 118-125
- Fluorouracil and dose-dense chemotherapy in adjuvant treatment of patients with early-stage breast cancer: an open-label, 2 x 2 factorial, randomised phase 3 trial.Lancet. 2015; 385: 1863-1872
- CircFBXL5 promotes the 5-FU resistance of breast cancer via modulating miR-216b/HMGA2 axis.Cancer Cell Int. 2021; 21: 384
- PLAC8 promotes adriamycin resistance via blocking autophagy in breast cancer.J Cell Mol Med. 2021; 25: 6948-6962
- Chromatin regulators mediate anthracycline sensitivity in breast cancer.Nat Med. 2019; 25: 1721-1727
- Tumor microenvironment and epithelial mesenchymal transition as targets to overcome tumor multidrug resistance.Drug Resist Updat. 2020; 53100715
- Interferon regulatory factor-8 regulates bone metabolism by suppressing osteoclastogenesis.Nat Med. 2009; 15: 1066-1071
- RANKL/RANK system-based mechanism for breast cancer bone metastasis and related therapeutic strategies.Front Cell Dev Biol. 2020; 8: 76
- Regulation of cancer cell migration and bone metastasis by RANKL.Nature. 2006; 440: 692-696
- Expression of receptor activator of NFkB (RANK) drives stemness and resistance to therapy in ER+HER2- breast cancer.Oncotarget. 2020; 11: 1714-1728
- A protein conjugation system essential for autophagy.Nature. 1998; 395: 395-398
- Autophagy inhibition enhances vorinostat-induced apoptosis via ubiquitinated protein accumulation.J Cell Mol Med. 2010; 14: 2448-2459
- Autophagy supports breast cancer stem cell maintenance by regulating IL6 secretion.Mol Cancer Res. 2015; 13: 651-658
- STAT3-mediated autophagy dependence identifies subtypes of breast cancer where autophagy inhibition can be efficacious.Cancer Res. 2014; 74: 2579-2590
- The interleukin-6/signal transducer and activator of transcription-3/cystathionine gamma-lyase axis deciphers the transformation between the sensitive and resistant phenotypes of breast cancer cells.Drug Metab Dispos. 2021; 49: 985-994
- Inhibition of STAT3-ferroptosis negative regulatory axis suppresses tumor growth and alleviates chemoresistance in gastric cancer.Redox Biol. 2022; 52102317
- JAK-STAT signalling controls cancer stem cell properties including chemotherapy resistance in myxoid liposarcoma.Int J Cancer. 2019; 145: 435-449
- Stattic inhibits RANKL-mediated osteoclastogenesis by suppressing activation of STAT3 and NF-kappaB pathways.Int Immunopharmacol. 2018; 58: 136-144
Published online: February 06, 2023
Accepted: January 31, 2023
Received in revised form: January 30, 2023
Received: August 3, 2022
Publication stageIn Press Journal Pre-Proof
© 2023 Published by Elsevier Inc.