Ozone Induces BEL7402 Cell Apoptosis by Increasing Reactive Oxygen Species Production and Activating JNK

Overview

Journal Ann Transl Med

Publisher AME Publishing Company

Date 2021 Sep 17

PMID 34532394

Citations 3

Authors

Shuiying Tang

Bihong Xu

Jincheng Li

Meifeng Zhong

Ziyang Hong

Wei Zhao

Tao Zeng

Xiaofeng He

Affiliations

Soon will be listed here.

Abstract

Background: Oxidative stress is an important factor in the modulation of both tumorigenesis and anticancer responses. Ozone (O) is a strong oxidant that causes redox reactions and exerts anticancer effects in various types of cancer cells. However, the pathways involved in O-induced cell death are not well understood.

Methods: human hepatocellular carcinoma (HCC) BEL7402 cells were treated with various O concentrations to evaluate O cytotoxicity by Cell Counting Kit-8 (CCK-8) assay and flow cytometry. The regulatory mechanisms were analyzed by western blot analysis. , an HCC model was established to evaluate the inhibition of HCC with O treatment.

Results: cells treated with O exhibited a round and small morphology with nuclear shrinkage and fragmentation. The CCK-8 assay confirmed the potent cytotoxic activity of O against BEL7402 cells (IC value of 5 µg/mL). Acridine orange/ethidium bromide (AO/EB) staining revealed apoptosis of BEL7402 cells after O treatment. Flow cytometry analysis showed that S phase cell cycle arrest and apoptosis increased with O exposure. In addition, O3 exposure reduced the mitochondrial membrane potential (ΔΨm) and induced reactive oxygen species (ROS) accumulation. Western blot analysis showed that O exposure reduced B-cell lymphoma 2 (BCL-2) expression and increased cleaved poly ADP-ribose polymerase (PARP), cytochrome C (Cyt-C), caspase-3, caspase-9, and p-JNK expression. , treatment with intratumor injection O (20 µg/mL) inhibited HCC growth.

Conclusions: Overall, our findings showed that O induces BEL7402 cell apoptosis via the intrinsic mitochondria-dependent pathway. Therefore, O has therapeutic potential for HCC.

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References

Mondal A, Bennett L . Resveratrol enhances the efficacy of sorafenib mediated apoptosis in human breast cancer MCF7 cells through ROS, cell cycle inhibition, caspase 3 and PARP cleavage. Biomed Pharmacother. 2016; 84:1906-1914. DOI: 10.1016/j.biopha.2016.10.096. View

Schenk R, Strasser A, Dewson G . BCL-2: Long and winding path from discovery to therapeutic target. Biochem Biophys Res Commun. 2017; 482(3):459-469. DOI: 10.1016/j.bbrc.2016.10.100. View

Zhang X, Wang X, Wu T, Li B, Liu T, Wang R . Isoliensinine induces apoptosis in triple-negative human breast cancer cells through ROS generation and p38 MAPK/JNK activation. Sci Rep. 2015; 5:12579. PMC: 4518223. DOI: 10.1038/srep12579. View

Sung H, Ferlay J, Siegel R, Laversanne M, Soerjomataram I, Jemal A . Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021; 71(3):209-249. DOI: 10.3322/caac.21660. View

Huang Y, Zhu D, Chen X, Chen Q, Luo Z, Liu C . Curcumin enhances the effects of irinotecan on colorectal cancer cells through the generation of reactive oxygen species and activation of the endoplasmic reticulum stress pathway. Oncotarget. 2017; 8(25):40264-40275. PMC: 5522238. DOI: 10.18632/oncotarget.16828. View

Chen J, Liu X, Chen K, Hu X, Xiang B, Yuan W . Sorafenib with or without concurrent transarterial chemoembolization in hepatocellular carcinoma: a cautionary comment of STAH trial. Transl Cancer Res. 2022; 9(1):400-401. PMC: 8797615. DOI: 10.21037/tcr.2019.12.51. View

Hsieh C, Kuo P, Hsu Y, Huang Y, Tsai E, Hsu Y . Arctigenin, a dietary phytoestrogen, induces apoptosis of estrogen receptor-negative breast cancer cells through the ROS/p38 MAPK pathway and epigenetic regulation. Free Radic Biol Med. 2013; 67:159-70. DOI: 10.1016/j.freeradbiomed.2013.10.004. View

Nie P, Hu W, Zhang T, Yang Y, Hou B, Zou Z . Synergistic Induction of Erlotinib-Mediated Apoptosis by Resveratrol in Human Non-Small-Cell Lung Cancer Cells by Down-Regulating Survivin and Up-Regulating PUMA. Cell Physiol Biochem. 2015; 35(6):2255-71. DOI: 10.1159/000374030. View

Bocci V, Borrelli E, Travagli V, Zanardi I . The ozone paradox: ozone is a strong oxidant as well as a medical drug. Med Res Rev. 2009; 29(4):646-82. DOI: 10.1002/med.20150. View

10.

Cannizzaro A, Verga Falzacappa C, Martinelli M, Misiti S, Brunetti E, Bucci B . O(2/3) exposure inhibits cell progression affecting cyclin B1/cdk1 activity in SK-N-SH while induces apoptosis in SK-N-DZ neuroblastoma cells. J Cell Physiol. 2007; 213(1):115-25. DOI: 10.1002/jcp.21097. View

11.

Llovet J, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc J . Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008; 359(4):378-90. DOI: 10.1056/NEJMoa0708857. View

12.

Simonetti V, Quagliariello V, Giustetto P, Franzini M, Iaffaioli R . Association of Ozone with 5-Fluorouracil and Cisplatin in Regulation of Human Colon Cancer Cell Viability: In Vitro Anti-Inflammatory Properties of Ozone in Colon Cancer Cells Exposed to Lipopolysaccharides. Evid Based Complement Alternat Med. 2018; 2017:7414083. PMC: 5868048. DOI: 10.1155/2017/7414083. View

13.

Gorrini C, Harris I, Mak T . Modulation of oxidative stress as an anticancer strategy. Nat Rev Drug Discov. 2013; 12(12):931-47. DOI: 10.1038/nrd4002. View

14.

Wei C, Chen P, Chau G, Lee R, Chen P, Huo T . Comparison of prognosis between surgical resection and transarterial chemoembolization for patients with solitary huge hepatocellular carcinoma. Ann Transl Med. 2020; 8(5):238. PMC: 7154415. DOI: 10.21037/atm.2019.12.157. View

15.

Yang J, Tang J, Liu Y, Wang H, Lee S, Yen C . Roe Protein Hydrolysates of Giant Grouper (Epinephelus lanceolatus) Inhibit Cell Proliferation of Oral Cancer Cells Involving Apoptosis and Oxidative Stress. Biomed Res Int. 2016; 2016:8305073. PMC: 4852358. DOI: 10.1155/2016/8305073. View

16.

Schmidt S, Follmann M, Malek N, Manns M, Greten T . Critical appraisal of clinical practice guidelines for diagnosis and treatment of hepatocellular carcinoma. J Gastroenterol Hepatol. 2011; 26(12):1779-86. PMC: 8369833. DOI: 10.1111/j.1440-1746.2011.06891.x. View

17.

Di Mauro R, Cantarella G, Bernardini R, Di Rosa M, Barbagallo I, Distefano A . The Biochemical and Pharmacological Properties of Ozone: The Smell of Protection in Acute and Chronic Diseases. Int J Mol Sci. 2019; 20(3). PMC: 6387239. DOI: 10.3390/ijms20030634. View

18.

Scassellati C, Carlo Galoforo A, Bonvicini C, Esposito C, Ricevuti G . Ozone: a natural bioactive molecule with antioxidant property as potential new strategy in aging and in neurodegenerative disorders. Ageing Res Rev. 2020; 63:101138. PMC: 7428719. DOI: 10.1016/j.arr.2020.101138. View

19.

Karlic H, Kucera H, Metka M, Schonbauer M, Soregi G . [Effect of ozone and ionizing radiation on an in vitro model--a pilot study of 4 gynecologic tumors]. Strahlenther Onkol. 1987; 163(1):37-42. View

20.

Zaman S, Wang R, Gandhi V . Targeting the apoptosis pathway in hematologic malignancies. Leuk Lymphoma. 2013; 55(9):1980-92. PMC: 4152229. DOI: 10.3109/10428194.2013.855307. View