Cantharidin inhibits cell proliferation and induces apoptosis through G2/M phase cell cycle arrest in hepatocellular carcinoma stem cells

Le,Ai-Ping; Zhang,Lun-Li; Liu,Wei; Shi,Yu-Fei

doi:10.3892/or.2016.4684

Cantharidin inhibits cell proliferation and induces apoptosis through G2/M phase cell cycle arrest in hepatocellular carcinoma stem cells

Authors:
- Ai-Ping Le
- Lun-Li Zhang
- Wei Liu
- Yu-Fei Shi
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Affiliations: Department of Blood Transfusion, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Infectious Diseases, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Published online on: March 17, 2016 https://doi.org/10.3892/or.2016.4684
Pages: 2970-2976

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Abstract

The present study was designed to investigate the effect of cantharidin on cell proliferation, ability of selfrenewal, cell cycle arrest and induction of apoptosis in HepG2 hepatocellular carcinoma stem cells (HCSCs). It was observed that cantharidin treatment exhibited dose- and time-dependent inhibitory effect on the viability of HCSCs. The inhibition of cell viability by cantharidin in HepG2 CD133+ and parental cells was significant at the concentration 5 and 15 µM, respectively after 48 h. Cantharidin treatment inhibited the self-renewal ability of the HCSCs and the expression of β-catenin and cyclin D1. Flow cytometry revealed that cantharidin treatment at 5 µM concentration significantly increased the cell population in G2/M phase and decreased the population in the G1 phase. Cantharidin treatment in the HCSCs for 48 h increased expression of histone H2AX, Myt1, cyclin A2, cyclin B1, p53 and cdc2 (Tyr15) phosphorylation significantly compared to the parental cells. Exposure of the HCSCs to cantharidin for 48 h at a concentration of 5 µM caused a significant increase in the proportion of apoptotic cells. Therefore, cantharidin is a promising agent for the hepatocellular carcinoma treatment.

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1	Ma S, Chan KW and Guan XY: In search of liver cancer stem cells. Stem Cell Rev. 4:179–192. 2008. View Article : Google Scholar : PubMed/NCBI
2	Tomuleasa C, Soritau O, Rus-Ciuca D, Pop T, Todea D, Mosteanu O, Pintea B, Foris V, Susman S, Kacsó G, et al: Isolation and characterization of hepatic cancer cells with stem-like properties from hepatocellular carcinoma. J Gastrointestin Liver Dis. 19:61–67. 2010.PubMed/NCBI
3	Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C and De Maria R: Identification and expansion of human colon-cancer-initiating cells. Nature. 445:1111152007. View Article : Google Scholar
4	Lee TK, Castilho A, Ma S and Ng IO: Liver cancer stem cells: Implications for a new therapeutic target. Liver Int. 29:955–965. 2009. View Article : Google Scholar : PubMed/NCBI
5	Mackillop WJ, Ciampi A, Till JE and Buick RN: A stem cell model of human tumor growth: Implications for tumor cell clonogenic assays. J Natl Cancer Inst. 70:9–16. 1983.PubMed/NCBI
6	Gupta PB, Chaffer CL and Weinberg RA: Cancer stem cells: Mirage or reality? Nat Med. 15:1010–1012. 2009. View Article : Google Scholar : PubMed/NCBI
7	Chiba T, Kita K, Zheng YW, Yokosuka O, Saisho H, Iwama A, Nakauchi H and Taniguchi H: Side population purified from hepatocellular carcinoma cells harbors cancer stem cell-like properties. Hepatology. 44:240–251. 2006. View Article : Google Scholar : PubMed/NCBI
8	Curry SC, Carlton MW and Raschke RA: Prevention of fetal and maternal cyanide toxicity from nitroprusside with coinfusion of sodium thiosulfate in gravid ewes. Anesth Analg. 84:1121–1126. 1997.PubMed/NCBI
9	Honkanen RE: Cantharidin, another natural toxin that inhibits the activity of serine/threonine protein phosphatases types 1 and 2A. FEBS Lett. 330:283–286. 1993. View Article : Google Scholar : PubMed/NCBI
10	Clarke PR, Hoffmann I, Draetta G and Karsenti E: Dephosphorylation of cdc25-C by a type-2A protein phosphatase: Specific regulation during the cell cycle in Xenopus egg extracts. Mol Biol Cell. 4:397–411. 1993. View Article : Google Scholar : PubMed/NCBI
11	Chen YN, Chen JC, Yin SC, Wang GS, Tsauer W, Hsu SF and Hsu SL: Effector mechanisms of norcantharidin-induced mitotic arrest and apoptosis in human hepatoma cells. Int J Cancer. 100:158–165. 2002. View Article : Google Scholar : PubMed/NCBI
12	Kok SH, Cheng SJ, Hong CY, Lee JJ, Lin SK, Kuo YS, Chiang CP and Kuo MY: Norcantharidin-induced apoptosis in oral cancer cells is associated with an increase of proapoptotic to antiapoptotic protein ratio. Cancer Lett. 217:43–52. 2005. View Article : Google Scholar
13	Huan SK, Lee HH, Liu DZ, Wu CC and Wang CC: Cantharidin-induced cytotoxicity and cyclooxygenase 2 expression in human bladder carcinoma cell line. Toxicology. 223:136–143. 2006. View Article : Google Scholar : PubMed/NCBI
14	Williams LA, Möller W, Merisor E, Kraus W and Rösner H: In vitro anti-proliferation/cytotoxic activity of cantharidin (Spanish Fly) and related derivatives. West Indian Med J. 52:10–13. 2003.PubMed/NCBI
15	Yi SN, Wass J, Vincent P and Iland H: Inhibitory effect of norcantharidin on K562 human myeloid leukemia cells in vitro. Leuk Res. 15:883–886. 1991. View Article : Google Scholar : PubMed/NCBI
16	Naujokat C and Steinhart R: Salinomycin as a drug for targeting human cancer stem cells. J Biomed Biotechnol. 2012:9506582012. View Article : Google Scholar : PubMed/NCBI
17	Wang Z, Li Y, Ahmad A, Azmi AS, Kong D, Banerjee S and Sarkar FH: Targeting miRNAs involved in cancer stem cell and EMT regulation: An emerging concept in overcoming drug resistance. Drug Resist Updat. 13:109–118. 2010. View Article : Google Scholar : PubMed/NCBI
18	Liu J, Kopecková P, Bühler P, Wolf P, Pan H, Bauer H, Elsässer-Beile U and Kopeček J: Biorecognition and subcellular trafficking of HPMA copolymeranti PSMA antibody conjugates by prostate cancer cells. Mol Pharm. 6:9599702009. View Article : Google Scholar
19	Mizrak D, Brittan M and Alison M: CD133: Molecule of the moment. J Pathol. 214:3–9. 2008. View Article : Google Scholar
20	Li L, Ying J, Li H, Zhang Y, Shu X, Fan Y, Tan J, Cao Y, Tsao SW, Srivastava G, et al: The human cadherin 11 is a proapoptotic tumor suppressor modulating cell stemness through Wnt/betacatenin signaling and silenced in common carcinomas. Oncogene. 31:390139122012. View Article : Google Scholar
21	Booher RN, Holman PS and Fattaey A: Human Myt1 is a cell cycle-regulated kinase that inhibits Cdc2 but not Cdk2 activity. J Biol Chem. 272:22300–22306. 1997. View Article : Google Scholar : PubMed/NCBI
22	Liu F, Stanton JJ, Wu Z and PiwnicaWorms H: The human Myt1 kinase preferentially phosphorylates Cdc2 on threonine 14 and localizes to the endoplasmic reticulum and Golgi complex. Mol Cell Biol. 17:5715831997. View Article : Google Scholar
23	Parker LL and Piwnica-Worms H: Inactivation of the p34cdc2-cyclin B complex by the human WEE1 tyrosine kinase. Science. 257:195519571992. View Article : Google Scholar
24	Raff MC, Barres BA, Burne JF, Coles HS, Ishizaki Y and Jacobson MD: Programmed cell death and the control of cell survival: Lessons from the nervous system. Science. 257:1955–1957. 1992.