Effect of Nitazoxanide on Human Osteosarcoma Cells and Its Underlying Mechanism
YE Cai-hong1, WEI Meng-qi1, WANG Yu-ping2, LUO Jin-yong1*
1. College of Medical Laboratory Science, Chongqing Medical University, Key Laboratory of Clinical Laboratory Diagnostics,Ministry of Education, Chongqing 400016, China; 2. Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
Abstract：OBJECTIVE To investigate the effect of nitazoxanide(NTZ) on osteosarcoma cells 143B and explore its underlying mechanism based on network pharmacology. METHODS The Swiss Target Prediction database and CTD database were used to obtain the drug target of NTZ, and the target genes of osteosarcoma were collected from Gene Cards database, then the drug-disease intersection genes were obtained by the intersection. The interaction network was constructed using STRING database and visualized with Cytoscape to obtain the drug-osteosarcoma-target protein regulatory network. Through calculating the degree value of each node, the first 20 genes were selected as key genes. Through GO and KEGG signaling pathway enrichment analysis in David database, the potential action pathway of the drug was predicted. In vitro experiments were validated. Human osteosarcoma cells were treated with 0,10,20,30,40 μmol·L-1 NTZ and 0. 05% DMSO,respectively. The effect of NTZ on the proliferation of osteosarcoma 143B cells was detected by crystal violet staining and MTT assay. Wound-healing assay and Transwell invasion assay were used to observe the migration and invasion ability of 143B cells after NTZ treatment. Apoptosis was detected by flow cytometry and Hoechst 33258 staining. The Protein level was detected by Western blot. RESULTS There are 78 potential targets of NTZ in osteosarcoma. GO enrichment analysis involved 306 items,and KEGG enrichment analysis obtained 80 items,involving tumor pathways,cell cycle signaling pathways,ErbB signaling pathways,etc. The results showed that NTZ could inhibit the proliferation,migration and invasion of osteosarcoma cells. Meanwhile,apoptosis was induced by NTZ,and significantly inhibit the phosphorylation of ERK1/2,a key molecule of ERK1/2 signaling pathway. CONCLUSION The effect of NTZ on osteosarcoma involves multi-target and multi-process,which has an inhibitory effect on osteosarcoma,and can inhibit the activation of ERK1/2 signaling pathway.
叶彩红, 魏梦琪, 王雨平, 罗进勇. 硝唑尼特对骨肉瘤的影响及机制研究[J]. 中国药学杂志, 2022, 57(10): 808-816.
YE Cai-hong, WEI Meng-qi, WANG Yu-ping, LUO Jin-yong. Effect of Nitazoxanide on Human Osteosarcoma Cells and Its Underlying Mechanism. Chinese Pharmaceutical Journal, 2022, 57(10): 808-816.
YOSHIDA A. Osteosarcoma: old and new challenges[J]. Surg Pathol Clin, 2021, 14(4):567-583.
[2]
GILL J, GORLICK R. Advancing therapy for osteosarcoma[J]. Nat Rev Clin Oncol, 2021, 18(10):609-624.
[3]
GOORIN A M, SCHWARTZENTRUBER D J, DEVIDAS M, et al. Presurgical chemotherapy compared with immediate surgery and adjuvant chemotherapy for nonmetastatic osteosarcoma: Pediatric Oncology Group Study POG-8651[J]. J Clin Oncol, 2003, 21(8):1574-1580.
[4]
KEMPF-BIELACK B, BIELACK S S, JÜRGENS H, et al. Osteosarcoma relapse after combined modality therapy: an analysis of unselected patients in the cooperative osteosarcoma study group(COSS)[J]. J Clin Oncol, 2005, 23(3):559-568.
[5]
ROSSIGNOL J F, BRÉCHOT C. A pilot clinical trial of nitazoxanide in the treatment of chronic hepatitis B[J]. Hepatol Commun, 2019, 3(6): 744-747.
[6]
WHITE C A, JR. Nitazoxanide: a new broad spectrum antiparasitic agent[J]. Expert Rev Anti Infect Ther, 2004, 2(1):43-49.
[7]
DI SANTO N, EHRISMAN J. A functional perspective of nitazoxanide as a potential anticancer drug[J]. Mutat Res, 2014, 768: 16-21. Doi:10.1016.j.mrfmmm.2014.05.005
[8]
QU Y, OLSEN J R, YUAN X, et al. Small molecule promotes β-catenin citrullination and inhibits Wnt signaling in cancer[J]. Nat Chem Biol, 2018, 14(1): 94-101.
[9]
GUO Y J, PAN W W, LIU S B, et al. ERK/MAPK signalling pathway and tumorigenesis[J]. Exp Ther Med, 2020, 19(3):1997-2007.
[10]
DE SOUZA A, TORRES L R, LIMA L R P, et al. Inhibition of Brazilian ZIKV strain replication in primary human placental chorionic cells and cervical cells treated with nitazoxanide[J]. Braz J Infect Dis, 2020, 24(6): 505-516.
[11]
HANG C C, WU Q, SHI J S, et al. Component analysis of dendrobium nobile lindl. alkaloids and its inhibitory effect on the apoptosis of pc12 cell induced by aβ1-42[J]. Chin Pharm J (中国药学杂志), 2021, 56(13): 1059-1067.
[12]
BASSIOUNI W, ALI M A M, SCHULZ R. Multifunctional intracellular matrix metalloproteinases: implications in disease[J]. Febs J, 2021, 288(24):7162-7182.
[13]
YANG Y Q, WANG H H, XIE K X, et al. MAPK/ERK inhibits proliferation, apoptosis, migration and related signal transduction of pituitary tumor cells by inducing fibrin a phosphorylation[J]. Pract Pharm Clin Reme(实用药物与临床), 2021, 24(1): 6-10.
[14]
WU J, ZHANG C, CHEN L. MiR-511 mimic transfection inhibits the proliferation, invasion of osteosarcoma cells and reduces metastatic osteosarcoma tumor burden in nude mice via targeting MAPK1[J]. Cancer Biomark, 2019, 26(3): 343-351.
[15]
ZHANG W, LIU Y X, WAN M, et al. The inhibition mechanism of chrysin on the metastasis of human colon cancer hct-116 cells by regulating yap-mediated EMT process[J]. Chin Pharm J(中国药学杂志), 2020, 55(23): 1948-1954.
[16]
KUNZ P, SÄHR H, LEHNER B, et al. Elevated ratio of MMP2/MMP9 activity is associated with poor response to chemotherapy in osteosarcoma[J]. BMC Cancer, 2016, 16:223.
[17]
SI H, PENG C, LI J, et al. RNAi-mediated knockdown of ERK1/2 inhibits cell proliferation and invasion and increases chemosensitivity to cisplatin in human osteosarcoma U2-OS cells in vitro[J]. Int J Oncol, 2012, 40(4): 1291-1297.
2022, Vol. 57 
