Autophagy inhibition induces the repolarisation of tumour-associated macrophages and enhances chemosensitivity of laryngeal cancer cells to cisplatin in mice
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Tumour-associated macrophages (TAMs) are the key components in the tumour microenvironment. TAMs have two major subtypes, M1 and M2. M1 macrophages are tumour inhibitory, while M2 macrophages are tumour promotive. Repolarising TAMs from M2 to M1 is a promising strategy in cancer treatment. M1 and M2 macrophages were generated from murine bone marrow-derived macrophages (BMDMs). We found that chloroquine (CQ), an autophagy inhibitor, was able to repolarise M2 macrophages to the anti-tumour M1 phenotype. The repolarised macrophages demonstrated higher phagocytotic activity towards Hep-2 laryngeal tumour cells and re-sensitised Hep-2 cells to cisplatin (CDDP) treatment in vitro. While CQ did not demonstrate cytotoxicity to Hep-2 cells in vitro, CQ treatment reduced Hep-2 laryngeal tumour growth in vivo and improved CDDP treatment outcomes. It seems that CQ-induced M2-to-M1 macrophage repolarisation played an important role in tumour growth inhibition, and the CQ/CDDP combined therapy might have clinical potential in laryngeal cancer treatment.
KeywordsChloroquine (CQ) Tumour-associated macrophages (TAMs) Autophagy inhibition Macrophage repolarisation Hep-2 laryngeal cancer
American type culture collection
Bone marrow-derived macrophage
Dulbecco’s modified Eagle medium
Fetal bovine serum
Microtubule-associated protein 1 light chain 3
Recombinant human macrophage colony stimulating factor
Nitric oxide synthase
Phosphate buffer saline
Quantitative real-time polymerase chain reaction
YG, YF, XC, QW conducted the experiments, YG and YF analysed the data. YG and XP wrote the manuscript. XP conceived the study.
This study was funded by the Project of Medical and Health Technology Development Program in Shandong (No. 2017WSB04024).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Animal studies on C57BL/6 mice and nu/nu nude mice complied with the Guidelines for Animal Studies in Shandong University. Ethical Committee in Shandong Provincial Qianfoshan Hospital reviewed and approved this study (#SDQFSH-2017048).
C57BL/6 mice and nu/nu nude mice were purchased from Nanjing Model Animal Institute (Nanjing, China).
Cell line authentication
Human laryngeal cancer Hep-2 cell line and GFP-labelled Hep-2-GFP cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and cells were verified by A Short Tandem Repeat analysis.
- 3.Ni Y-H, Ding L, Huang X-F, Dong Y-c, Hu Q-G, Hou Y-Y (2015) Microlocalization of CD68(+) tumor-associated macrophages in tumor stroma correlated with poor clinical outcomes in oral squamous cell carcinoma patients. Tumor Biol 36:5291–5298. https://doi.org/10.1007/s13277-015-3189-5 CrossRefGoogle Scholar
- 5.Jinushi M, Chiba S, Yoshiyama H, Masutomi K, Kinoshita I, Dosaka-Akita H, Yagita H, Takaoka A, Tahara H (2011) Tumor-associated macrophages regulate tumorigenicity and anticancer drug responses of cancer stem/initiating cells. Proc Natl Acad Sci USA 108:12425–12430. https://doi.org/10.1073/pnas.1106645108 CrossRefPubMedGoogle Scholar
- 6.Mitchem JB, Brennan DJ, Knolhoff BL, Belt BA, Zhu Y, Sanford DE, Belaygorod L, Carpenter D, Collins L, Piwnica-Worms D, Hewitt S, Udupi GM, Gallagher WM, Wegner C, West BL, Wang-Gillam A, Goedegebuure P, Linehan DC, DeNardo DG (2013) Targeting tumor-infiltrating macrophages decreases tumor-initiating cells, relieves immunosuppression, and improves chemotherapeutic responses. Can Res 73:1128–1141. https://doi.org/10.1158/0008-5472.Can-12-2731 CrossRefGoogle Scholar
- 7.Nakasone ES, Askautrud HA, Kees T, Park J-H, Plaks V, Ewald AJ, Fein M, Rasch MG, Tan Y-X, Qiu J, Park J, Sinha P, Bissell MJ, Frengen E, Werb Z, Egeblad M (2012) Imaging tumor-stroma interactions during chemotherapy reveals contributions of the microenvironment to resistance. Cancer Cell 21:488–503. https://doi.org/10.1016/j.ccr.2012.02.017 CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Colegio OR, Ngoc-Quynh C, Szabo AL, Chu T, Rhebergen AM, Jairam V, Cyrus N, Brokowski CE, Eisenbarth SC, Phillips GM, Cline GW, Phillips AJ, Medzhitov R (2014) Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 513:559. https://doi.org/10.1038/nature13490 CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Rodell CB, Arlauckas SP, Cuccarese MF, Garris CS, Ahmed RLMS, Kohler RH, Pittet MJ, Weissleder R (2018) TLR7/8-agonist-loaded nanoparticles promote the polarization of tumour-associated macrophages to enhance cancer immunotherapy. Nat Biomed Eng 2:578. https://doi.org/10.1038/s41551-018-0236-8 CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Yang S, Wang X, Contino G, Liesa M, Sahin E, Ying H, Bause A, Li Y, Stommel JM, Dell’Antonio G, Mautner J, Tonon G, Haigis M, Shirihai OS, Doglioni C, Bardeesy N, Kimmelman AC (2011) Pancreatic cancers require autophagy for tumor growth. Genes Dev 25:717–729. https://doi.org/10.1101/gad.2016111 CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Kulkarni A, Chandrasekar V, Natarajan SK, Ramesh A, Pandey P, Nirgud J, Bhatnagar H, Ashok D, Ajay AK, Sengupta S (2018) A designer self-assembled supramolecule amplifies macrophage immune responses against aggressive cancer. Nat Biomed Eng 2:589. https://doi.org/10.1038/s41551-018-0254-6 CrossRefPubMedPubMedCentralGoogle Scholar
- 23.Li R, Zhou R, Wang H, Li W, Pan M, Yao X, Zhan W, Yang S, Xu L, Ding Y, Zhao L (2019) Gut microbiota-stimulated cathepsin K secretion mediates TLR4-dependent M2 macrophage polarization and promotes tumor metastasis in colorectal cancer. Cell Death Differ. https://doi.org/10.1038/s41418-019-0312-y CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Kurahara H, Takao S, Maemura K, Mataki Y, Kuwahata T, Maeda K, Sakoda M, Iino S, Ishigami S, Ueno S, Shinchi H, Natsugoe S (2013) M2-polarized tumor-associated macrophage infiltration of regional lymph nodes is associated with nodal lymphangiogenesis and occult nodal involvement in pN0 pancreatic cancer. Pancreas 42:155–159. https://doi.org/10.1097/MPA.0b013e318254f2d1 CrossRefPubMedGoogle Scholar
- 25.Chen Y, Wen H, Zhou C, Su Q, Lin Y, Xie Y, Huang Y, Qiu Q, Lin J, Huang X, Tan W, Min C, Wang C (2019) TNF-alpha derived from M2 tumor-associated macrophages promotes epithelial-mesenchymal transition and cancer stemness through the Wnt/beta-catenin pathway in SMMC-7721 hepatocellular carcinoma cells. Exp Cell Res. https://doi.org/10.1016/j.yexcr.2019.03.005 CrossRefPubMedGoogle Scholar
- 28.Pyonteck SM, Akkari L, Schuhmacher AJ, Bowman RL, Sevenich L, Quail DF, Olson OC, Quick ML, Huse JT, Teijeiro V, Setty M, Leslie CS, Oei Y, Pedraza A, Zhang J, Brennan CW, Sutton JC, Holland EC, Daniel D, Joyce JA (2013) CSF-1R inhibition alters macrophage polarization and blocks glioma progression. Nat Med 19:1264. https://doi.org/10.1038/nm.3337 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Guo B, Li L, Guo J, Liu A, Wu J, Wang H, Shi J, Pang D, Cao Q (2017) M2 tumor-associated macrophages produce interleukin-17 to suppress oxaliplatin-induced apoptosis in hepatocellular carcinoma. Oncotarget 8:44465–44476. https://doi.org/10.18632/oncotarget.17973 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Degtyarev M, De Maziere A, Orr C, Lin J, Lee BB, Tien JY, Prior WW, van Dijk S, Wu H, Gray DC, Davis DP, Stern HM, Murray LJ, Hoeflich KP, Klumperman J, Friedman LS, Lin K (2008) Akt inhibition promotes autophagy and sensitizes PTEN-null tumors to lysosomotropic agents. J Cell Biol 183:101–116. https://doi.org/10.1083/jcb.200801099 CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Hamed HA, Yacoub A, Park MA, Eulitt P, Sarkar D, Dimitriev IP, Chen C-S, Grant S, Curiel DT, Fisher PB, Dent P (2010) OSU-03012 enhances Ad.mda-7-induced GBM cell killing via ER stress and autophagy and by decreasing expression of mitochondrial protective proteins. Cancer Biol Ther 9:526–536. https://doi.org/10.4161/cbt.9.7.11116 CrossRefPubMedPubMedCentralGoogle Scholar