Bufadienolides induce p53-mediated apoptosis in esophageal squamous cell carcinoma cells in vitro and in vivo

Lin,Shaohuan; Lv,Junhong; Peng,Panli; Cai,Changqing; Deng,Jianming; Deng,Haihong; Li,Xuejun; Tang,Xinyue

doi:10.3892/ol.2017.7457

Bufadienolides induce p53-mediated apoptosis in esophageal squamous cell carcinoma cells in vitro and in vivo

Authors:
- Shaohuan Lin
- Junhong Lv
- Panli Peng
- Changqing Cai
- Jianming Deng
- Haihong Deng
- Xuejun Li
- Xinyue Tang
View Affiliations

Affiliations: Thoracic Surgeons Department, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong 510317, P.R. China, Oncology No. 2 Department, Guangdong Second Provincial General Hospital, Guangzhou, Guangdong 510317, P.R. China
Published online on: November 21, 2017 https://doi.org/10.3892/ol.2017.7457
Pages: 1566-1572
Copyright: © Lin 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

Bufadienolides are a type of cardiotonic steroids isolated from the skin and parotid venom glands of the toad Bufo bufo gargarizans Cantor, and exhibit wide‑spectrum anticancer activities. However, the effects and mechanisms of bufadienolides on esophageal squamous cell carcinoma (ESCC) cells remain unknown. In the present study, the anticancer activities of two bufadienolides, bufotalin and bufalin, were examined in vitro and in vivo. The results demonstrated that bufotalin and bufalin effectively inhibited the viability of ESCC cells, with half‑maximal inhibitory concentration (IC50) values of 0.8‑3.6 µM. However, bufotalin and bufalin exhibited lower toxicity towards Het‑1A human esophageal squamous cells, indicating their high selectivity towards cancer cells. Mechanistic studies revealed that bufotalin effectively induced ESCC cell apoptosis, as characterized by DNA fragmentation and nuclear condensation, which was primarily mediated through activation of caspase family members. In addition, treatment of ESCC cells with bufotalin markedly activated tumor protein p53 (p53) phosphorylation. Transfection of cells with p53 small interfering RNA markedly inhibited bufotalin‑induced p53 phosphorylation and significantly inhibited bufotalin‑induced cell apoptosis. Furthermore, bufotalin demonstrated in vivo anticancer efficacy in a tumor‑bearing nude mice model, where bufotalin effectively inhibited Eca‑109 xenograft tumor growth in a time‑ and dose‑dependent manner, through activation of the p53 signaling pathway. Collectively, the results from the present study suggested that bufadienolides exert anticancer effects against ESCC by regulating the p53 signaling pathway.

View Figures

View References

1	Song G, Cheng L, Chao Y, Yang K and Liu Z: Emerging nanotechnology and advanced materials for cancer radiation therapy. Adv Mater. 29:2017. View Article : Google Scholar
2	Humbert PO, Verona R, Trimarchi JM, Rogers C, Dandapani S and Lees JA: E2f3 is critical for normal cellular proliferation. Genes Dev. 14:690–703. 2000.PubMed/NCBI
3	Chang Y, He L, Li Z, Zeng L, Song Z, Li P, Chan L, You Y, Yu XF, Chu PK and Chen T: Designing core-shell gold and selenium nanocomposites for cancer radiochemotherapy. Acs Nano. 11:4848–4858. 2017. View Article : Google Scholar : PubMed/NCBI
4	Singh SK, Banerjee S, Acosta EP, Lillard JW and Singh R: Resveratrol induces cell cycle arrest and apoptosis with docetaxel in prostate cancer cells via a p53/p21(WAF1/CIP1) and p27(KIP1) pathway. Oncotarget. 8:17216–17228. 2017.PubMed/NCBI
5	Liong M, Lu J, Kovochich M, Xia T, Ruehm SG, Nel AE, Tamanoi F and Zink JI: Multifunctional inorganic nanoparticles for imaging, targeting, and drug delivery. ACS Nano. 2:889–896. 2008. View Article : Google Scholar : PubMed/NCBI
6	Oelkrug C, Hartke M and Schubert A: Mode of action of anticancer peptides (ACPs) from amphibian origin. Anticancer Res. 35:635–643. 2015.PubMed/NCBI
7	Chen T and Wong YS: Selenocystine induces apoptosis of A375 human melanoma cells by activating ROS-mediated mitochondrial pathway and p53 phosphorylation. Cell Mol Life Sci. 65:2763–2775. 2008. View Article : Google Scholar : PubMed/NCBI
8	Chen T and Wong YS: Selenocystine induces caspase-independent apoptosis in MCF-7 human breast carcinoma cells with involvement of p53 phosphorylation and reactive oxygen species generation. Int J Biochem Cell Biol. 41:666–676. 2009. View Article : Google Scholar : PubMed/NCBI
9	Qi FH, Li AY, Lv H, Zhao L, Li JJ, Gao B and Tang W: Apoptosis-inducing effect of cinobufacini, Bufo bufo gargarizans Cantor skin extract, on human hepatoma cell line BEL-7402. Drug Discov Ther. 2:339–343. 2008.PubMed/NCBI
10	Chen Y, Qiao L, Yu B, Li G, Liu C, Ji L and Chao H: Mitochondria-specific phosphorescent imaging and tracking in living cells with an AIPE-active iridium(III) complex. Chem Commun (Camb). 49:11095–11097. 2013. View Article : Google Scholar : PubMed/NCBI
11	Wang Z, Wen J, Zhang J, Ye M and Guo D: Simultaneous determination of four bufadienolides in human liver by high-performance liquid chromatography. Biomed Chromatogr. 18:318–322. 2004. View Article : Google Scholar : PubMed/NCBI
12	Wen L, Huang Y, Xie X, Huang W, Yin J, Lin W, Jia Q and Zeng W: Anti-inflammatory and antinociceptive activities of bufalin in rodents. Mediators Inflamm. 2014:1718392014. View Article : Google Scholar : PubMed/NCBI
13	Wang DL, Qi FH, Xu HL, Inagaki Y, Orihara Y, Sekimizu K, Kokudo N, Wang FS and Tang W: Apoptosis-inducing activity of compounds screened and characterized from cinobufacini by bioassay-guided isolation. Mol Med Rep. 3:717–22. 2010.PubMed/NCBI
14	Xu GX and Wang TT: Apoptosis of lens epithelial cells induced by cinobufagin in vitro. Int J Ophthalmol. 3:128–131. 2010.PubMed/NCBI
15	Zhai XF, Fang FF, Liu Q, Meng YB, Guo YY and Chen Z: MiR-181a contributes to bufalin-induced apoptosis in PC-3 prostate cancer cells. BMC Complement Altern Med. 13:3252013. View Article : Google Scholar : PubMed/NCBI
16	Wang F, Zhang D, Zhang Q, Chen Y, Zheng D, Hao L, Duan C, Jia L, Liu G and Liu Y: Synergistic effect of folate-mediated targeting and verapamil-mediated P-gp inhibition with paclitaxel-polymer micelles to overcome multi-drug resistance. Biomaterials. 32:9444–9456. 2011. View Article : Google Scholar : PubMed/NCBI
17	Chen YY, Lu HF, Hsu SC, Kuo CL, Chang SJ, Lin JJ, Wu PP, Liu JY, Lee CH, Chung JG and Chang JB: Bufalin inhibits migration and invasion in human hepatocellular carcinoma SK-Hep1 cells through the inhibitions of NF-kB and matrix metalloproteinase-2/−9-signaling pathways. Environ Toxicol. 30:74–82. 2015. View Article : Google Scholar : PubMed/NCBI
18	Wang W, Shi A and Fan Z: Apoptosis of T-47D cells induced by cinobufacini via a caspase-3-dependent manner. Chem Res Chinese Univer. 30:108–113. 2014. View Article : Google Scholar
19	Li M, Yu X, Guo H, Sun L, Wang A, Liu Q, Wang X and Li J: Bufalin exerts antitumor effects by inducing cell cycle arrest and triggering apoptosis in pancreatic cancer cells. Tumour Biol. 35:2461–2471. 2014. View Article : Google Scholar : PubMed/NCBI
20	Zhang DM, Liu JS, Deng LJ, Chen MF, Yiu A, Cao HH, Tian HY, Fung KP, Kurihara H, Pan JX and Ye WC: Arenobufagin, a natural bufadienolide from toad venom, induces apoptosis and autophagy in human hepatocellular carcinoma cells through inhibition of PI3K/Akt/mTOR pathway. Carcinogenesis. 34:1331–1342. 2013. View Article : Google Scholar : PubMed/NCBI
21	Luo H, Wang F, Bai Y, Chen T and Zheng W: Selenium nanoparticles inhibit the growth of HeLa and MDA-MB-231 cells through induction of S phase arrest. Colloids Surf B Biointerfaces. 94:304–308. 2012. View Article : Google Scholar : PubMed/NCBI
22	Yang F, Tang Q, Zhong X, Bai Y, Chen T, Zhang Y, Li Y and Zheng W: Surface decoration by Spirulina polysaccharide enhances the cellular uptake and anticancer efficacy of selenium nanoparticles. Int J Nanomedicine. 7:835–844. 2012.PubMed/NCBI
23	Zhang Y, Li X, Huang Z, Zheng W, Fan C and Chen T: Enhancement of cell permeabilization apoptosis-inducing activity of selenium nanoparticles by ATP surface decoration. Nanomedicine. 9:74–84. 2013. View Article : Google Scholar : PubMed/NCBI
24	Maiese K, Chong ZZ, Shang YC and Wang S: Targeting disease through novel pathways of apoptosis and autophagy. Expert Opin Ther Targets. 16:1203–1214. 2012. View Article : Google Scholar : PubMed/NCBI
25	Chen T and Wong YS: Selenocystine induces apoptosis of A375 human melanoma cells by activating ROS-mediated mitochondrial pathway and p53 phosphorylation. Cell Mol Life Sci. 65:2763–2775. 2008. View Article : Google Scholar : PubMed/NCBI
26	Chen T and Wong YS: Selenocystine induces caspase-independent apoptosis in MCF-7 human breast carcinoma cells with involvement of p53 phosphorylation and reactive oxygen species generation. Int J Biochem Cell Biol. 41:666–676. 2009. View Article : Google Scholar : PubMed/NCBI
27	Riedl SJ and Shi Y: Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol. 5:897–907. 2004. View Article : Google Scholar : PubMed/NCBI
28	He L, Lai H and Chen T: Dual-function nanosystem for synergetic cancer chemo-/radiotherapy through ROS-mediated signaling pathways. Biomaterials. 51:30–42. 2015. View Article : Google Scholar : PubMed/NCBI
29	Li M, Lai L, Zhao Z and Chen T: Aquation is a crucial activation step for anticancer action of ruthenium(II) polypyridyl complexes to trigger cancer cell apoptosis. Chem Asian J. 11:310–320. 2016. View Article : Google Scholar : PubMed/NCBI
30	Xie Q, Lan G, Zhou Y, Huang J, Liang Y, Zheng W, Fu X, Fan C and Chen T: Strategy to enhance the anticancer efficacy of X-ray radiotherapy in melanoma cells by platinum complexes, the role of ROS-mediated signaling pathways. Cancer Lett. 354:58–67. 2014. View Article : Google Scholar : PubMed/NCBI
31	Fridman JS and Lowe SW: Control of apoptosis by p53. Oncogene. 22:9030–9040. 2003. View Article : Google Scholar : PubMed/NCBI
32	He L, Huang Y, Zhu H, Pang G, Zheng W, Wong YS and Chen T: Cancer-targeted monodisperse mesoporous silica nanoparticles as carrier of ruthenium polypyridyl complexes to enhance theranostic effects. Adv Funct Mater. 24:2754–2763. 2014. View Article : Google Scholar