Microwave hyperthermia enhances the sensitivity of lung cancer cells to gemcitabine through reactive oxygen species‑induced autophagic death

Yang,Yang; Yang,Chun‑Li; Zhao,Zhen‑Jiang; Zuo,Xiao‑Xiao; Liang,Tian‑Song; Yang,Ya; Ma,Sheng‑Lin; Yang,Dao‑Ke

doi:10.3892/or.2019.7085

Microwave hyperthermia enhances the sensitivity of lung cancer cells to gemcitabine through reactive oxygen species‑induced autophagic death

Authors:
- Yang Yang
- Chun‑Li Yang
- Zhen‑Jiang Zhao
- Xiao‑Xiao Zuo
- Tian‑Song Liang
- Ya Yang
- Sheng‑Lin Ma
- Dao‑Ke Yang
View Affiliations

Affiliations: Department of Radiation Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China, Gynecology and Obstetrics, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Emergency Department, Children's Hospital Affiliated to Zhengzhou University, Zhengzhou, Henan 450053, P.R. China, Center for Translational Medicine, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China
Published online on: March 22, 2019 https://doi.org/10.3892/or.2019.7085
Pages: 3100-3110

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The pleiotropic effects of hyperthermia on cancer cells have been well documented, and microwave hyperthermia (MWHT) has been widely applied for multifarious cancer treatment. However, the mechanisms underlying the anticancer effect of MWHT combined with gemcitabine (GEM) remain poorly understood. The aim of the present study was to investigate the role of autophagy in the thermo‑chemotherapy of human squamous cell lung carcinoma cells. It was observed that MWHT combined with GEM potently suppressed the viability of NCI‑H2170 and NCI‑H1703 cells, and induced G0/G1 cell cycle arrest. Notably, MWHT with GEM induced autophagy, as indicated by the formation of autophagic vacuoles, downregulation of p62 and upregulation of light chain 3‑II. It was further demonstrated that the autophagy was due to the production of reactive oxygen species (ROS), whereas N‑acetyl cysteine, an ROS scavenger, attenuated the level of autophagy. However, when the autophagy inhibitor 3‑methyladenine was used, there was no significant change in the production of ROS. Furthermore, it was observed that MWHT combined with GEM downregulated the protein expression levels of phosphoinositide 3‑kinase (PI3K), phosphorylated (p)‑PI3K, protein kinase B (AKT), p‑AKT, mammalian target of rapamycin (mTOR), p‑mTOR, phosphorylated S6 (pS6) and p70 S6 kinase, which are associated with autophagy. In addition, the results demonstrated that ROS served as an upstream mediator of PI3K/AKT/mTOR signaling. In light of these findings, the present study provides original insights into the molecular mechanisms underlying the cell death induced by MWHT combined with GEM, and this may be a promising approach for the treatment of human squamous cell lung carcinoma.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2015. CA Cancer J Clin. 65:5–29. 2015. View Article : Google Scholar : PubMed/NCBI
2	Heist RS, Mino-Kenudson M, Sequist LV, Tammireddy S, Morrissey L, Christiani DC, Engelman JA and Iafrate AJ: FGFR1 amplification in squamous cell carcinoma of the lung. J Thorac Oncol. 7:1775–1780. 2012. View Article : Google Scholar : PubMed/NCBI
3	Derman BA, Mileham KF, Bonomi PD, Batus M and Fidler MJ: Treatment of advanced squamous cell carcinoma of the lung: A review. Transl Lung Cancer Res. 4:524–532. 2015.PubMed/NCBI
4	Sun Y, Wu YL, Zhou CC, Zhang L, Zhang L, Liu XY, Yu SY, Jiang GL, Li K, Qin SK, et al: Second-line pemetrexed versus docetaxel in Chinese patients with locally advanced or metastatic non-small cell lung cancer: A randomized, open-label study. Lung Cancer. 79:143–150. 2013. View Article : Google Scholar : PubMed/NCBI
5	Palumbo R, Sottotetti F, Trifirò G, Piazza E, Ferzi A, Gambaro A, Spinapolice EG, Pozzi E, Tagliaferri B, Teragni C, et al: Nanoparticle albumin-bound paclitaxel (nab-paclitaxel) as second-line chemotherapy in HER2-negative, taxane-pretreated metastatic breast cancer patients: Prospective evaluation of activity, safety, and quality of life. Drug Des Devel Ther. 9:2189–2199. 2015. View Article : Google Scholar : PubMed/NCBI
6	Duan J, Wang Z, Bai H, An T, Zhuo M, Wu M, Wang Y, Yang L and Wang J: Epidermal growth factor receptor variant III mutation in Chinese patients with squamous cell cancer of the lung. Thorac Cancer. 6:319–326. 2015. View Article : Google Scholar : PubMed/NCBI
7	Hurwitz M and Stauffer P: Hyperthermia, radiation and chemotherapy: The role of heat in multidisciplinary cancer care. Semin Oncol. 41:714–729. 2014. View Article : Google Scholar : PubMed/NCBI
8	Salahi S, Maccarini PF, Rodrigues DB, Etienne W, Landon CD, Inman BA, Dewhirst MW and Stauffer PR: Miniature microwave applicator for murine bladder hyperthermia studies. Int J Hyperthermia. 28:456–465. 2012. View Article : Google Scholar : PubMed/NCBI
9	Asano M, Tanaka S, Sakaguchi M, Matsumura H, Yamaguchi T, Fujita Y and Tabuse K: Normothermic microwave irradiation induces death of HL-60 cells through Heat-independent apoptosis. Sci Rep. 7:114062017. View Article : Google Scholar : PubMed/NCBI
10	Li C, Li C, Ge H, Liang M, Ma G, Ling L, Pan H, Gong H, Xie H, Ding Q, et al: Technical analysis of US imaging for precise microwave ablation for benign breast tumours. Int J Hyperthermia. 34:1179–1185. 2018. View Article : Google Scholar : PubMed/NCBI
11	Han JB, Kong FW, Ding H, Zhang YF, Liu JM, Wei Q, Hu L, Zhao L, Xu CJ and Yi YX: Hepatectomy combined with microwave ablation of the spleen for treatment of hepatocellular carcinoma complicated with splenomegaly: A retrospective study. Mol Clin Oncol. 6:204–208. 2017. View Article : Google Scholar : PubMed/NCBI
12	Kouloulias V, Triantopoulou S, Vrouvas J, Gennatas K, Oozounoglou N, Kouvaris J, Karaiskos P, Aggelakis P, Antypas C, Zygogianni A, et al: Combined chemoradiotherapy with local microwave hyperthermia for treatment of T3N0 laryngeal carcinoma: A retrospective study with long-term follow-up. Acta Otorhinolaryngol Ital. 34:167–173. 2014.PubMed/NCBI
13	Othman T, Goto S, Lee JB, Taimura A, Matsumoto T and Kosaka M: Hyperthermic enhancement of the apoptotic and antiproliferative activities of paclitaxel. Pharmacology. 62:208–212. 2001. View Article : Google Scholar : PubMed/NCBI
14	Adachi S, Kokura S, Okayama T, Ishikawa T, Takagi T, Handa O, Naito Y and Yoshikawa T: Effect of hyperthermia combined with gemcitabine on apoptotic cell death in cultured human pancreatic cancer cell lines. Int J Hyperthermia. 25:210–219. 2009. View Article : Google Scholar : PubMed/NCBI
15	Zhao YY, Wu Q, Wu ZB, Zhang JJ, Zhu LC, Yang Y, Ma SL and Zhang SR: Microwave hyperthermia promotes caspase-3-dependent apoptosis and induces G₂/M checkpoint arrest via the ATM pathway in non-small cell lung cancer cells. Int J Oncol. 53:539–550. 2018.PubMed/NCBI
16	Yang Y, Zhao Y, Ma S and Yang D: Microwave hyperthermia combined with gemcitabine inhibits proliferation and induces apoptosis of human lung squamous carcinoma cells. Zhongguo Fei Ai Za Zhi. 21:805–814. 2018.(In Chinese). PubMed/NCBI
17	White EJ, Martin V, Liu JL, Klein SR, Piya S, Gomez-Manzano C, Fueyo J and Jiang H: Autophagy regulation in cancer development and therapy. Am J Cancer Res. 1:362–372. 2011.PubMed/NCBI
18	Rosenfeldt MT and Ryan KM: The multiple roles of autophagy in cancer. Carcinogenesis. 32:955–963. 2011. View Article : Google Scholar : PubMed/NCBI
19	Wang H, Zhang T, Sun W, Wang Z, Zuo D, Zhou Z, Li S, Xu J, Yin F, Hua Y, et al: Erianin induces G2/M-phase arrest, apoptosis, and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells in vitro and in vivo. Cell Death Dis. 7:e22472016. View Article : Google Scholar : PubMed/NCBI
20	Ba MC, Long H, Wang S, Wu YB, Zhang BH, Yan ZF, Yu FH and Cui SZ: Hyperthermia enhances radiosensitivity of colorectal cancer cells through ROS inducing autophagic cell death. J Cell Biochem. 119:3763–3774. 2018. View Article : Google Scholar : PubMed/NCBI
21	Chen F, Wang CC, Kim E and Harrison LE: Hyperthermia in combination with oxidative stress induces autophagic cell death in HT-29 colon cancer cells. Cell Biol Int. 32:715–723. 2008. View Article : Google Scholar : PubMed/NCBI
22	Wang Z, Cai F, Chen X, Luo M, Hu L and Lu Y: The role of mitochondria-derived reactive oxygen species in hyperthermia-induced platelet apoptosis. PLoS One. 8:e750442013. View Article : Google Scholar : PubMed/NCBI
23	Hou CH, Lin FL, Hou SM and Liu JF: Hyperthermia induces apoptosis through endoplasmic reticulum and reactive oxygen species in human osteosarcoma cells. Int J Mol Sci. 15:17380–17395. 2014. View Article : Google Scholar : PubMed/NCBI
24	Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, Agholme L, Agnello M, Agostinis P, Aguirre-Ghiso JA, et al: Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 8:445–544. 2012. View Article : Google Scholar : PubMed/NCBI
25	Martinotti S, Ranzato E, Parodi M, Vitale M and Burlando B: Combination of ascorbate/epigallocatechin-3-gallate/gemcitabine synergistically induces cell cycle deregulation and apoptosis in mesothelioma cells. Toxicol Appl Pharmacol. 274:35–41. 2014. View Article : Google Scholar : PubMed/NCBI
26	Furusawa Y, Iizumi T, Fujiwara Y, Zhao QL, Tabuchi Y, Nomura T and Kondo T: Inhibition of checkpoint kinase 1 abrogates G2/M checkpoint activation and promotes apoptosis under heat stress. Apoptosis. 17:102–112. 2012. View Article : Google Scholar : PubMed/NCBI
27	Michalakis J, Georgatos SD, Romanos J, Koutala H, Georgoulias V, Tsiftsis D and Theodoropoulos PA: Micromolar taxol, with or without hyperthermia induces mitotic catastrophe and cell necrosis in HeLa cells. Cancer Chemother Pharmacol. 56:615–622. 2005. View Article : Google Scholar : PubMed/NCBI
28	Haveman J, Rietbroek RC, Geerdink A, Van Rijn J and Bakker PJ: Effect of hyperthermia on the cytotoxicity of 2′,2′-difluorodeoxycytidine (gemcitabine) in cultured SW1573 cells. Int J Cancer. 62:627–630. 1995. View Article : Google Scholar : PubMed/NCBI
29	Ba MC, Long H, Cui SZ, Gong YF, Yan ZF, Wang S and Wu YB: Mild hyperthermia enhances sensitivity of gastric cancer cells to chemotherapy through reactive oxygen species-induced autophagic death. Tumor Biol. 39:10104283177119522017. View Article : Google Scholar
30	Zhang L, Wang H, Xu J, Zhu J and Ding K: Inhibition of cathepsin S induces autophagy and apoptosis in human glioblastoma cell lines through ROS-mediated PI3K/AKT/mTOR/p70S6K and JNK signaling pathways. Toxico Lett. 228:248–259. 2014. View Article : Google Scholar
31	Dewaele M, Maes H and Agostinis P: ROS-mediated mechanisms of autophagy stimulation and their relevance in cancer therapy. Autophagy. 6:838–854. 2010. View Article : Google Scholar : PubMed/NCBI
32	Xiong Y, Yepuri G, Forbiteh M, Yu Y, Montani JP, Yang Z and Ming XF: ARG2 impairs endothelial autophagy through regulation of MTOR and PRKAA/AMPK signaling in advanced atherosclerosis. Autophagy. 10:2223–2238. 2014. View Article : Google Scholar : PubMed/NCBI
33	Zhang Q, Zhu H, Xu X, Li L, Tan H and Cai X: Inactivated Sendai virus induces apoptosis and autophagy via the PI3K/Akt/mTOR/p70S6K pathway in human nonsmall cell lung cancer cells. Biochem Biophys Res Commun. 465:64–70. 2015. View Article : Google Scholar : PubMed/NCBI
34	Papadimitrakopoulou V: Development of PI3K/AKT/mTOR pathway inhibitors and their application in personalized therapy for non-small-cell lung cancer. J Thorac Oncol. 7:1315–1326. 2012. View Article : Google Scholar : PubMed/NCBI
35	Zhou M, Shen S, Zhao X and Gong X: Luteoloside induces G0/G1 arrest and pro-death autophagy through the ROS-mediated AKT/mTOR/p70S6K signalling pathway in human non-small cell lung cancer cell lines. Biochem Biophys Res Commun. 494:263–269. 2017. View Article : Google Scholar : PubMed/NCBI