Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

Okamoto,Hiroki; Yoshikawa,Kazuhiro; Shimada,Akiko; Sano,Rui; Inukai,Daisuke; Yamanaka,Shunpei; Suzuki,Susumu; Ueda,Ryuzo; Ueda,Hiromi; Fujimoto,Yasushi; Ogawa,Tetsuya

doi:10.3892/or.2023.8591

Artesunate and cisplatin synergistically inhibit HNSCC cell growth and promote apoptosis with artesunate‑induced decreases in Rb and phosphorylated Rb levels

Authors:
- Hiroki Okamoto
- Kazuhiro Yoshikawa
- Akiko Shimada
- Rui Sano
- Daisuke Inukai
- Shunpei Yamanaka
- Susumu Suzuki
- Ryuzo Ueda
- Hiromi Ueda
- Yasushi Fujimoto
- Tetsuya Ogawa
View Affiliations

Affiliations: Department of Otorhinolaryngology‑Head and Neck Surgery, Aichi Medical University School of Medicine, Nagakute, Aichi 480‑1195, Japan, Division of Biobank, Research Creation Support Center, Aichi Medical University, Nagakute, Aichi 480‑1195, Japan, Division of Research Creation, Research Creation Support Center, Aichi Medical University, Nagakute, Aichi 480‑1195, Japan, Department of Tumor Immunology, Aichi Medical University School of Medicine, Nagakute, Aichi 480‑1195, Japan
Published online on: June 19, 2023 https://doi.org/10.3892/or.2023.8591
Article Number: 154
Copyright: © Okamoto 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

In the treatment of head and neck cancer, cisplatin is often used as a therapeutic agent; however, its efficacy is limited and it can cause renal dysfunction as an adverse effect. For this reason, the use of cisplatin is limited in elderly patients with reduced renal function. Recently, artemisinin, which was developed as an antimalarial drug, was found to have antitumor effects and is effective in combination with other anticancer drugs. In the present study, the antitumor effects of artemisinin and its derivatives as well as their combination with cisplatin and iron on head and neck squamous cell carcinoma cell lines, were investigated. Cell viability was determined by a cell viability assay, the cell cycle was analyzed by flow cytometry, cell death was assessed with annexin V and propidium iodide staining, and western blotting was used to analyze retinoblastoma protein (Rb), phosphorylated (p‑)Rb, and other cell cycle‑associated molecules. A total of four artemisinin compounds were examined and it was found that artesunate and dihydroartemisinin had a significant inhibitory effect on growth. It was also identified that the combination of artesunate, cisplatin, and iron inhibited cell proliferation and caused S/G2‑M cell cycle arrest. In addition, western blotting of Rb, a molecule involved in the cell cycle, showed that artesunate induced the loss of not only Rb but also p‑Rb. These results suggested that artesunate is a useful drug in combination with cisplatin.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Morton RP, Rugman F, Dorman EB, Stoney PJ, Wilson JA, McCormick M, Veevers A and Stell PM: Cisplatinum and bleomycin for advanced or recurrent squamous cell carcinoma of the head and neck: A randomised factorial phase III controlled trial. Cancer Chemother Pharmacol. 15:283–289. 1985. View Article : Google Scholar : PubMed/NCBI
3	Kish JA, Weaver A, Jacobs J, Cummings G and Al-Sarraf M: Cisplatin and 5-fluorouracil infusion in patients with recurrent and disseminated epidermoid cancer of the head and neck. Cancer. 53:1819–1824. 1984. View Article : Google Scholar : PubMed/NCBI
4	Kish J, Drelichman A, Jacobs J, Hoschner J, Kinzie J, Loh J, Weaver A and Al-Sarraf M: Clinical trial of cisplatin and 5-FU infusion as initial treatment for advanced squamous cell carcinoma of the head and neck. Cancer Treat Rep. 66:471–474. 1982.PubMed/NCBI
5	Ogawa T, Niho S, Nagai S, Kojima T, Nishimura Y, Ohe Y, Kondo N, Yamaguchi T, Endo K, Izumi K and Minami H: Moderate renal dysfunction may not require a cisplatin dose reduction: A retrospective study of cancer patients with renal impairment. Int J Clin Oncol. 18:977–982. 2013. View Article : Google Scholar : PubMed/NCBI
6	Rosenthal PJ: Artesunate for the treatment of severe falciparum malaria. N Engl J Med. 358:1829–1836. 2008. View Article : Google Scholar : PubMed/NCBI
7	Ho WE, Peh HY, Chan TK and Wong WS: Artemisinins: Pharmacological actions beyond anti-malarial. Pharmacol Ther. 142:126–139. 2014. View Article : Google Scholar : PubMed/NCBI
8	Sun WC, Han JX, Yang WY, Deng DA and Yue XF: Antitumor activities of 4 derivatives of artemisic acid and artemisinin B in vitro. Acta Pharmacol Sin. 13:541–543. 1992.
9	Efferth T, Dunstan H, Sauerbrey A, Miyachi H and Chitambar CR: The anti-malarial artesunate is also active against cancer. Int J Oncol. 18:767–773. 2001.PubMed/NCBI
10	Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO, Hengstler JG, Halatsch ME, Volm M, Tew KD, et al: Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol. 64:382–394. 2003. View Article : Google Scholar : PubMed/NCBI
11	Våtsveen TK, Myhre MR, Steen CB, Wälchli S, Lingjærde OC, Bai B, Dillard P, Theodossiou TA, Holien T, Sundan A, et al: Artesunate shows potent anti-tumor activity in B-cell lymphoma. J Hematol Oncol. 11:232018. View Article : Google Scholar : PubMed/NCBI
12	Berte N, Lokan S, Eich M, Kim E and Kaina B: Artesunate enhances the therapeutic response of glioma cells to temozolomide by inhibition of homologous recombination and senescence. Oncotarget. 7:67235–67250. 2016. View Article : Google Scholar : PubMed/NCBI
13	Hou J, Wang D, Zhang R and Wang H: Experimental therapy of hepatoma with artemisinin and its derivatives: In vitro and in vivo activity, chemosensitization, and mechanisms of action. Clin Cancer Res. 14:5519–5530. 2008. View Article : Google Scholar : PubMed/NCBI
14	Roh JL, Kim EH, Jang H and Shin D: Nrf2 inhibition reverses the resistance of cisplatin-resistant head and neck cancer cells to artesunate-induced ferroptosis. Redox Biol. 11:254–262. 2017. View Article : Google Scholar : PubMed/NCBI
15	Jia J, Qin Y, Zhang L, Guo C, Wang Y, Yue X and Qian J: Artemisinin inhibits gallbladder cancer cell lines through triggering cell cycle arrest and apoptosis. Mol Med Rep. 13:4461–4468. 2016. View Article : Google Scholar : PubMed/NCBI
16	Chen S, Gan S, Han L, Li X, Xie X, Zou D and Sun H: Artesunate induces apoptosis and inhibits the proliferation, stemness, and tumorigenesis of leukemia. Ann Transl Med. 8:7672020. View Article : Google Scholar : PubMed/NCBI
17	Dell'Eva R, Pfeffer U, Vené R, Anfosso L, Forlani A, Albini A and Efferth T: Inhibition of angiogenesis in vivo and growth of Kaposi's sarcoma xenograft tumors by the anti-malarial artesunate. Biochem Pharmacol. 68:2359–2366. 2004. View Article : Google Scholar : PubMed/NCBI
18	Kim SJ, Kim MS, Lee JW, Lee CH, Yoo H, Shin SH, Park MJ and Lee SH: Dihydroartemisinin enhances radiosensitivity of human glioma cells in vitro. J Cancer Res Clin Oncol. 132:129–135. 2006. View Article : Google Scholar : PubMed/NCBI
19	Efferth T: From ancient herb to modern drug: Artemisia annua and artemisinin for cancer therapy. Semin Cancer Biol. 46:65–83. 2017. View Article : Google Scholar : PubMed/NCBI
20	Wong YK, Xu C, Kalesh KA, He Y, Lin Q, Wong WSF, Shen HM and Wang J: Artemisinin as an anticancer drug: Recent advances in target profiling and mechanisms of action. Med Res Rev. 37:1492–1517. 2017. View Article : Google Scholar : PubMed/NCBI
21	Xu C, Zhang H, Mu L and Yang X: Artemisinins as anticancer drugs: Novel therapeutic approaches, molecular mechanisms, and clinical trials. Front Pharmacol. 11:5298812020. View Article : Google Scholar : PubMed/NCBI
22	Nishimura K, Tsuchiya Y, Okamoto H, Ijichi K, Gosho M, Fukayama M, Yoshikawa K, Ueda H, Bradford CR, Carey TE and Ogawa T: Identification of chemoresistant factors by protein expression analysis with iTRAQ for head and neck carcinoma. Br J Cancer. 111:799–806. 2014. View Article : Google Scholar : PubMed/NCBI
23	Muramatsu H, Sumitomo M, Morinaga S, Kajikawa K, Kobayashi I, Nishikawa G, Kato Y, Watanabe M, Zennami K, Kanao K, et al: Targeting lactate dehydrogenase-A promotes docetaxel-induced cytotoxicity predominantly in castration-resistant prostate cancer cells. Oncol Rep. 42:224–230. 2019.PubMed/NCBI
24	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
25	Zhang JL, Wang Z, Hu W, Chen SS, Lou XE and Zhou HJ: DHA regulates angiogenesis and improves the efficiency of CDDP for the treatment of lung carcinoma. Microvasc Res. 87:14–24. 2013. View Article : Google Scholar : PubMed/NCBI
26	Razavi A, Nouri HR, Mehrabian F and Mirshafiey A: Treatment of experimental nephrotic syndrome with artesunate. Int J Toxicol. 26:373–380. 2007. View Article : Google Scholar : PubMed/NCBI
27	Wang SJ, Gao Y, Chen H, Kong R, Jiang HC, Pan SH, Xue DB, Bai XW and Sun B: Dihydroartemisinin inactivates NF-kappaB and potentiates the anti-tumor effect of gemcitabine on pancreatic cancer both in vitro and in vivo. Cancer Lett. 293:99–108. 2010. View Article : Google Scholar : PubMed/NCBI
28	Zhao F, Vakhrusheva O, Markowitsch SD, Slade KS, Tsaur I, Cinatl J Jr, Michaelis M, Efferth T, Haferkamp A and Juengel E: Artesunate impairs growth in cisplatin-resistant bladder cancer cells by cell cycle arrest, apoptosis and autophagy induction. Cells. 9:26432020. View Article : Google Scholar : PubMed/NCBI
29	Tran KQ, Tin AS and Firestone GL: Artemisinin triggers a G1 cell cycle arrest of human Ishikawa endometrial cancer cells and inhibits cyclin-dependent kinase-4 promoter activity and expression by disrupting nuclear factor-κB transcriptional signaling. Anticancer Drugs. 25:270–281. 2014. View Article : Google Scholar : PubMed/NCBI
30	Ji Y, Zhang YC, Pei LB, Shi LL, Yan JL and Ma XH: Anti-tumor effects of dihydroartemisinin on human osteosarcoma. Mol and Cell Biochem. 351:99–108. 2011. View Article : Google Scholar : PubMed/NCBI
31	Chen K, Shou LM, Lin F, Duan WM, Wu MY, Xie X, Xie YF, Li W and Tao M: Artesunate induces G2/M cell cycle arrest through autophagy induction in breast cancer cells. Anticancer Drugs. 25:652–662. 2014. View Article : Google Scholar : PubMed/NCBI
32	An B and Dou QP: Cleavage of retinoblastoma protein during apoptosis: An interleukin 1 beta-converting enzyme-like protease as candidate. Cancer Res. 56:438–442. 1996.PubMed/NCBI
33	Chen WD, Otterson GA, Lipkowitz S, Khleif SN, Coxon AB and Kaye FJ: Apoptosis is associated with cleavage of a 5 kDa fragment from RB which mimics dephosphorylation and modulates E2F binding. Oncogene. 14:1243–1248. 1997. View Article : Google Scholar : PubMed/NCBI
34	Tan X and Wang JY: The caspase-RB connection in cell death. Trends Cell Biol. 8:116–120. 1998. View Article : Google Scholar : PubMed/NCBI
35	Fan HN, Zhu MY, Peng SQ, Zhu JS, Zhang J and Qu GQ: Dihydroartemisinin inhibits the growth and invasion of gastric cancer cells by regulating cyclin D1-CDK4-Rb signaling. Pathol Res Pract. 216:1527952020. View Article : Google Scholar : PubMed/NCBI
36	Almasan A, Yin Y, Kelly RE, Lee EY, Bradley A, Li W, Bertino JR and Wahl GM: Deficiency of retinoblastoma protein leads to inappropriate S-phase entry, activation of E2F-responsive genes, and apoptosis. Proc Natl Acad Sci USA. 92:5436–5440. 1995. View Article : Google Scholar : PubMed/NCBI