AbstractObjective To observe the optimal time interval of multiple short-term hyperthermia in tongue squamous carcinoma Cal-27 cells, and investigate the role of Notch signaling pathway in hyperthermia-induced apoptosis of Cal-27 cells. Methods Cal-27 cells were placed into a 42℃ incubator for 45 min, and the second hyperthermia was performed at 6 h,12 h, 24 h, 48 h and 72 h later, respectively. Cell proliferation capacity was detected by CCK-8 assay at 0 h, 12 h and 24 h after the end of hyperthermia. After determining the optimal time interval of hyperthermia, multiple cycles of hyperthermia were performed, and the effects of hyperthermia on the proliferation and apoptosis of Cal-27 cells were detected by CCK-8 assay and flow cytometry, respectively. The changes in the expression of Notch1, Jagged1 and hairy and enhancer of split homolog-1 (Hes1) mRNA and proteins were analyzed by real-time reverse transcription PCR (qRT-PCR) and Western blot. All data were expressed as mean±SD. Two-group comparison was performed by one-way ANOVA. Results Hyperthermia at 42℃ for 45 min every 12 h consistently inhibited the proliferation of Cal-27 cells (P<0.05). Compared with the control group, hyperthermia in the experiment group significantly inhibited the proliferation and induced the apoptosis of Cal-27 cells after 7 cycles of hyperthermia (P<0.05), and the expression levels of Notch1, Jagged1 and Hes1 mRNA and proteins in the hyperthermia group were significantly down-regulated (all P<0.05). Conclusions The optimal time interval of hyperthermia to inhibit the proliferation of tongue squamous cells carcinoma Cal-27 cells is 12 h. Hyperthermia might induce the apoptosis of Cal-27 cells by inhibiting the Notch signaling pathway.
Fund:The Scientific and Technological Foundation of Gansu Province (20JR10FA670)
Corresponding Authors:
Wang Jing, Email: lzukqwj@126.com
Cite this article:
Lyu Yurong,Zheng He,Qi Jing et al. A preliminary study of apoptosis in tongue squamous cell carcinoma cells induced by hyperthermia via downregulation of Notch1/Jagged1/Hes1[J]. Chinese Journal of Radiation Oncology, 2023, 32(4): 347-352.
Lyu Yurong,Zheng He,Qi Jing et al. A preliminary study of apoptosis in tongue squamous cell carcinoma cells induced by hyperthermia via downregulation of Notch1/Jagged1/Hes1[J]. Chinese Journal of Radiation Oncology, 2023, 32(4): 347-352.
[1] Shi FZ, Xue DF, Jiang QK, et al.Glaucocalyxin A induces apoptosis and autophagy in tongue squamous cell carcinoma cells by regulating ROS[J]. Cancer Chemother Pharmacol, 2021,88(2):235-246. DOI: 10.1007/s00280- 021-04285-3. [2] Grégoire V, Lefebvre JL, Licitra L, et al. Squamous cell carcinoma of the head and neck: EHNS-ESMO-ESTRO clinical practice guidelines for diagnosis, treatment and follow-up[J]. Ann Oncol, 2010,21 Suppl 5:v184-186. DOI: 10.1093/annonc/mdq185. [3] D'Cruz AK, Vaish R, Kapre N, et al. Elective versus therapeutic neck dissection in node-negative oral cancer[J]. N Engl J Med, 2015,373(6):521-529. DOI: 10.1056/NEJMoa1506007. [4] Nagata M, Nakayama H, Tanaka T, et al.Overexpression of cIAP2 contributes to 5-FU resistance and a poor prognosis in oral squamous cell carcinoma[J]. Br J Cancer, 2011,105(9):1322-1330. DOI: 10.1038/bjc.2011.387. [5] Solomon B, Young RJ, Rischin D.Head and neck squamous cell carcinoma: genomics and emerging biomarkers for immunomodulatory cancer treatments[J]. Semin Cancer Biol, 2018,52(Pt 2):228-240. DOI: 10.1016/j.semcancer. 2018.01.008. [6] Hegyi G, Szigeti GP, Szász A.Hyperthermia versus oncothermia: cellular effects in complementary cancer therapy[J]. Evid Based Complement Alternat Med, 2013,2013:672873. DOI: 10.1155/2013/672873. [7] Raut CP, Evans DB, Crane CH, et al. Neoadjuvant therapy for respectable pancreatic cancer[J]. Surg Oncol Clin North Am, 2004, 13(4):639-661. DOI: 10.1016/j.soc. 2004.06.007. [8] Mouratidis PX, Rivens I, Ter Haar G.A study of thermal dose-induced autophagy, apoptosis and necroptosis in colon cancer cells[J]. Int J Hyperthermia, 2015,31(5):476-488. DOI: 10.3109/02656736.2015.1029995. [9] Reichrath J, Reichrath S.Notch pathway and inherited diseases: challenge and promise[J]. Adv Exp Med Biol, 2020,1218:159-187. DOI: 10.1007/978-3-030-34436- 8_9. [10] Zhu JN, Jiang L, Jiang JH, et al.Hepatocyte nuclear factor-1beta enhances the stemness of hepatocellular carcinoma cells through activation of the Notch pathway[J]. Sci Rep, 2017,7(1):4793. DOI: 10.1038/s41598-017- 04116-7. [11] Huang T, Zhou Y, Cheng AS, et al.NOTCH receptors in gastric and other gastrointestinal cancers: oncogenes or tumor suppressors?[J]. Mol Cancer, 2016,15(1):80. DOI: 10.1186/s12943-016-0566-7. [12] Scutigliani EM, Liang Y, Crezee H, et al. Modulating the heat stress response to improve hyperthermia-based anticancer treatments[J]. Cancers (Basel), 2021,13(6)DOI: 10.3390/cancers13061243. [13] Tyrrell D, Barrow I, Arthur J.Local hyperthermia benefits natural and experimental common colds[J]. BMJ, 1989,298(6683):1280-1283. DOI: 10.1136/bmj.298.6683.1280. [14] Robinson V, Brosseau L, Casimiro L, et al. Thermotherapy for treating rheumatoid arthritis[J]. Cochrane Database Syst Rev, 2002,(2):CD002826. DOI: 10.1002/14651858.CD002826. [15] Gao XH, Gao D, Sun XP, et al.Non-ablative controlled local hyperthermia for common warts[J]. Chin Med J (Engl), 2009,122(17):2061-2063. [16] Sheng LM, Ji YL, Wu QE, et al.Regional hyperthermia combined with radiotherapy for esophageal squamous cell carcinoma with supraclavicular lymph node metastasis[J]. Oncotarget, 2017,8(3):5339-5348. DOI: 10. 18632/oncotarget.14148. [17] Klimanov MY, Syvak LA, Orel VE, et al.Efficacy of combined regional inductive moderate hyperthermia and chemotherapy in patients with multiple liver metastases from breast cancer[J]. Technol Cancer Res Treat, 2018,17:1533033818806003. DOI: 10.1177/1533 033818806003. [18] Mallory M, Gogineni E, Jones GC, et al.Therapeutic hyperthermia: the old, the new, and the upcoming[J]. Crit Rev Oncol Hematol, 2016,97:56-64. DOI: 10.1016/j.critrevonc.2015.08.003. [19] 中国临床肿瘤学会肿瘤热疗专家委员会, 中日医学科技交流协会热疗专家委员会, 中华医学会放疗分会热疗学组. 肿瘤热疗中国专家共识[J].实用肿瘤杂志,2020,35(1):1-10. DOI: 10.13267/j.cnki.syzlzz.2020.01.001. Guidelines Working Committee of Chinese Society of Clinical Oncology, Sino Japan Science and Technology Association, Chinese Medical Association on Radiotherapy Branch Thermotherapy Specialized Committee. Chinese expert consensus on clinical application of tumor hyperthermia[J]. J of Pract Oncology,2020,35(1):1-10. DOI: 10.13267/j.cnki.syzlzz.2020. 01.001. [20] Li GC, Hahn GM.A proposed operational model of thermotolerance based on effects of nutrients and the initial treatment temperature[J]. Cancer Res, 1980,40(12):4501-4508. [21] Amorim FT, Fonseca IT, Machado-Moreira CA, et al.Insights into the role of heat shock protein 72 to whole-body heat acclimation in humans[J]. Temperature (Austin), 2015,2(4):499-505. DOI: 10.1080/23328940. 2015.1110655. [22] Braune EB, Lendahl U.Notch -- a goldilocks signaling pathway in disease and cancer therapy[J]. Discov Med, 2016,21(115):189-196. [23] Huang QF, Li JH, Zheng JH, et al.The carcinogenic role of the Notch signaling pathway in the development of hepatocellular carcinoma[J]. J Cancer, 2019,10(6):1570-1579. DOI: 10.7150/jca.26847. [24] Monticone G, Miele L.Notch pathway: a journey from Notching phenotypes to cancer immunotherapy[J]. Adv Exp Med Biol, 2021,1287:201-222. DOI: 10.1007/978-3-030-55031-8_13. [25] Wu-Chou YH, Hsieh CH, Liao CT, et al.NOTCH1 mutations as prognostic marker in oral squamous cell carcinoma[J]. Pathol Res Pract, 2021,223:153474. DOI: 10.1016/j.prp.2021.153474.
2023, Vol. 32 
