In vitro and in vivo antitumor effects of chloroquine on oral squamous cell carcinoma

Jia,Lihua; Wang,Juan; Wu,Tong; Wu,Jinan; Ling,Junqi; Cheng,Bin

doi:10.3892/mmr.2017.7342

In vitro and in vivo antitumor effects of chloroquine on oral squamous cell carcinoma

Authors:
- Lihua Jia
- Juan Wang
- Tong Wu
- Jinan Wu
- Junqi Ling
- Bin Cheng
View Affiliations

Affiliations: Department of Oral Medicine, Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China, Department of Oral and Maxillofacial Surgery, Affiliated Zhongshan Hospital, Sun Yat‑Sen University, Zhongshan, Guangdong 528403, P.R. China, Department of Endodontics, Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China
Published online on: August 23, 2017 https://doi.org/10.3892/mmr.2017.7342
Pages: 5779-5786
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Chloroquine, which is a widely used antimalarial drug, has been reported to exert anticancer activity in some tumor types; however, its potential effects on oral squamous cell carcinoma (OSCC) remain unclear. The present study aimed to explore the effects and possible underlying mechanisms of chloroquine against OSCC. MTT and clonogenic assays were conducted to evaluate the effects of chloroquine on the human OSCC cell lines SCC25 and CAL27. Cell cycle progression and apoptosis were detected using flow cytometry. Autophagy was monitored using microtubule-associated protein 1A/1B‑light chain 3 as an autophagosomal marker. In order to determine the in vivo antitumor effects of chloroquine on OSCC, a CAL27 xenograft model was used. The results demonstrated that chloroquine markedly inhibited the proliferation and the colony‑forming ability of both OSCC cell lines in a dose‑ and time‑dependent manner in vitro. Chloroquine also disrupted the cell cycle, resulting in the cell cycle arrest of CAL27 and SCC25 cells at G0/G1 phase, via downregulation of cyclin D1. In addition, chloroquine inhibited autophagy, and induced autophagosome and autolysosome accumulation in the cytoplasm, thus interfering with degradation; however, OSCC apoptosis was barely affected by chloroquine. The results of the in vivo study demonstrated that chloroquine effectively inhibited OSCC tumor growth in the CAL27 xenograft model. In conclusion, the present study reported the in vitro and in vivo antitumor effects of chloroquine on OSCC, and the results indicated that chloroquine may be considered a potent therapeutic agent against human OSCC.

View Figures

View References

1	Kimura T, Takabatake Y, Takahashi A and Isaka Y: Chloroquine in cancer therapy: A double-edged sword of autophagy. Cancer Res. 73:3–7. 2013. View Article : Google Scholar : PubMed/NCBI
2	Al-Bari MA: Chloroquine analogues in drug discovery: New directions of uses, mechanisms of actions and toxic manifestations from malaria to multifarious diseases. J Antimicrob Chemother. 70:1608–1621. 2015.PubMed/NCBI
3	Yang ZJ, Takabatake Y, Takahashi A and Isaka Y: The role of autophagy in cancer: Therapeutic implications. Mol Cancer Ther. 10:1533–1541. 2011. View Article : Google Scholar : PubMed/NCBI
4	Johnson NW, Jayasekara P and Amarasinghe AA: Squamous cell carcinoma and precursor lesions of the oral cavity: Epidemiology and aetiology. Periodontol 2000. 57:19–37. 2011. View Article : Google Scholar : PubMed/NCBI
5	Pignon JP, Bourhis J, Domenge C and Designé L: Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: Three meta-analyses of updated individual data. MACH-NC collaborative group. Meta-analysis of chemotherapy on head and neck cancer. Lancet. 355:949–955. 2000. View Article : Google Scholar : PubMed/NCBI
6	Sturgis EM, Moore BA, Glisson BS, Kies MS, Shin DM and Byers RM: Neoadjuvant chemotherapy for squamous cell carcinoma of the oral tongue in young adults: A case series. Head Neck. 27:748–756. 2005. View Article : Google Scholar : PubMed/NCBI
7	Freier K, Engel M, Lindel K, Flechtenmacher C, Mühling J, Hassfeld S and Hofele C: Neoadjuvant concurrent radiochemotherapy followed by surgery in advanced oral squamous cell carcinoma (OSCC): A retrospective analysis of 207 patients. Oral Oncol. 44:116–123. 2008. View Article : Google Scholar : PubMed/NCBI
8	Wedemeyer I, Kreppel M, Scheer M, Zöller JE, Büttner R and Drebber U: Histopathological assessment of tumour regression, nodal stage and status of resection margins determines prognosis in patients with oral squamous cell carcinoma treated with neoadjuvant radiochemotherapy. Oral Dis. 20:e81–e89. 2014. View Article : Google Scholar : PubMed/NCBI
9	Jiang PD, Zhao YL, Shi W, Deng XQ, Xie G, Mao YQ, Li ZG, Zheng YZ, Yang SY and Wei YQ: Cell growth inhibition, G2/M cell cycle arrest and apoptosis induced by chloroquine in human breast cancer cell line Bcap-37. Cell Physiol Biochem. 22:431–440. 2008. View Article : Google Scholar : PubMed/NCBI
10	Zheng Y, Zhao YL, Deng X, Yang S, Mao Y, Li Z, Jiang P, Zhao X and Wei Y: Chloroquine inhibits colon cancer cell growth in vitro and tumor growth in vivo via induction of apoptosis. Cancer Invest. 27:286–292. 2009. View Article : Google Scholar : PubMed/NCBI
11	Hu T, Li P, Luo Z, Chen X, Zhang J, Wang C, Chen P and Dong Z: Chloroquine inhibits hepatocellular carcinoma cell growth in vitro and in vivo. Oncol Rep. 35:43–49. 2016. View Article : Google Scholar : PubMed/NCBI
12	Fan C, Wang W, Zhao B, Zhang S and Miao J: Chloroquine inhibits cell growth and induces cell death in A549 lung cancer cells. Bioorg Med Chem. 14:3218–3222. 2006. View Article : Google Scholar : PubMed/NCBI
13	Geng Y, Kohli L, Klocke BJ and Roth KA: Chloroquine-induced autophagic vacuole accumulation and cell death in glioma cells is p53 independent. Neuro Oncol. 12:473–481. 2010.PubMed/NCBI
14	Lakhter AJ, Sahu RP, Sun Y, Kaufmann WK, Androphy EJ, Travers JB and Naidu SR: Chloroquine promotes apoptosis in melanoma cells by inhibiting BH3 domain-mediated PUMA degradation. J Invest Dermatol. 133:2247–2254. 2013. View Article : Google Scholar : PubMed/NCBI
15	Wang J, Jia L, Kuang Z, Wu T, Hong Y, Chen X, Leung WK, Xia J and Cheng B: The in vitro and in vivo antitumor effects of clotrimazole on oral squamous cell carcinoma. PLoS One. 9:e988852014. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Tanida I, Ueno T and Kominami E: LC3 conjugation system in mammalian autophagy. Int J Biochem Cell Biol. 36:2503–2518. 2004. View Article : Google Scholar : PubMed/NCBI
18	Pascolo S: Time to use a dose of Chloroquine as an adjuvant to anti-cancer chemotherapies. Eur J Pharmacol. 771:139–144. 2016. View Article : Google Scholar : PubMed/NCBI
19	Lefort S, Joffre C, Kieffer Y, Givel AM, Bourachot B, Zago G, Bieche I, Dubois T, Meseure D, Vincent-Salomon A, et al: Inhibition of autophagy as a new means of improving chemotherapy efficiency in high-LC3B triple-negative breast cancers. Autophagy. 10:2122–2142. 2014. View Article : Google Scholar : PubMed/NCBI
20	Liang DH, Choi DS, Ensor JE, Kaipparettu BA, Bass BL and Chang JC: The autophagy inhibitor chloroquine targets cancer stem cells in triple negative breast cancer by inducing mitochondrial damage and impairing DNA break repair. Cancer Lett. 376:249–258. 2016. View Article : Google Scholar : PubMed/NCBI
21	Choi DS, Blanco E, Kim YS, Rodriguez AA, Zhao H, Huang TH, Chen CL, Jin G, Landis MD, Burey LA, et al: Chloroquine eliminates cancer stem cells through deregulation of Jak2 and DNMT1. Stem Cells. 32:2309–2323. 2014. View Article : Google Scholar : PubMed/NCBI
22	Angadi PV and Krishnapillai R: Cyclin D1 expression in oral squamous cell carcinoma and verrucous carcinoma: Correlation with histological differentiation. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 103:e30–e35. 2007. View Article : Google Scholar : PubMed/NCBI
23	Zhong LP, Zhu DW, William WN Jr, Liu Y, Ma J, Yang CZ, Yang X, Wang LZ, Li J, Myers JN, et al: Elevated cyclin D1 expression is predictive for a benefit from TPF induction chemotherapy in oral squamous cell carcinoma patients with advanced nodal disease. Mol Cancer Ther. 12:1112–1121. 2013. View Article : Google Scholar : PubMed/NCBI
24	Zhao Y, Yu D, Li H, Nie P, Zhu Y, Liu S, Zhu M and Fang B: Cyclin D1 overexpression is associated with poor clinicopathological outcome and survival in oral squamous cell carcinoma in Asian populations: Insights from a meta-analysis. PLoS One. 9:e932102014. View Article : Google Scholar : PubMed/NCBI
25	Myo K, Uzawa N, Miyamoto R, Sonoda I, Yuki Y and Amagasa T: Cyclin D1 gene numerical aberration is a predictive marker for occult cervical lymph node metastasis in TNM Stage I and II squamous cell carcinoma of the oral cavity. Cancer. 104:2709–2716. 2005. View Article : Google Scholar : PubMed/NCBI
26	Zhao XG, Sun RJ, Yang XY, Liu DY, Lei DP, Jin T and Pan XL: Chloroquine-enhanced efficacy of cisplatin in the treatment of hypopharyngeal carcinoma in xenograft mice. PLoS One. 10:e01261472015. View Article : Google Scholar : PubMed/NCBI
27	Verschooten L, Barrette K, Van Kelst S, Romero N Rubio, Proby C, De Vos R, Agostinis P and Garmyn M: Autophagy inhibitor chloroquine enhanced the cell death inducing effect of the flavonoid luteolin in metastatic squamous cell carcinoma cells. PLoS One. 7:e482642012. View Article : Google Scholar : PubMed/NCBI
28	Quan HY, Quan HY, Zhou LJ, Li AD and Zhang ZB: Mechanism of chloroquine in promoting sensitivity of chemotherapeutics in oral squamous cell carcinoma CAL-27 cell line to cisplatin. Shanghai Kou Qiang Yi Xue. 24:30–36. 2015.(In Chinese). PubMed/NCBI
29	Amaravadi RK, Lippincott-Schwartz J, Yin XM, Weiss WA, Takebe N, Timmer W, DiPaola RS, Lotze MT and White E: Principles and current strategies for targeting autophagy for cancer treatment. Clin Cancer Res. 17:654–666. 2011. View Article : Google Scholar : PubMed/NCBI
30	Kapoor V, Paliwal D, Singh S Baskar, Mohanti BK and Das SN: Deregulation of Beclin 1 in patients with tobacco-related oral squamous cell carcinoma. Biochem Biophys Res Commun. 422:764–769. 2012. View Article : Google Scholar : PubMed/NCBI
31	Nomura H, Uzawa K, Yamano Y, Fushimi K, Ishigami T, Kouzu Y, Koike H, Siiba M, Bukawa H, Yokoe H, et al: Overexpression and altered subcellular localization of autophagy-related 16-like 1 in human oral squamous-cell carcinoma: Correlation with lymphovascular invasion and lymph-node metastasis. Hum Pathol. 40:83–91. 2009. View Article : Google Scholar : PubMed/NCBI