Bufalin induces apoptosis in the U‑2OS human osteosarcoma cell line via triggering the mitochondrial pathway

Chen,Ya; Li,Meng; Li,Zhongji; Gao,Peng; Zhou,Xiao; Zhang,Jianxin

doi:10.3892/mmr.2015.4583

Bufalin induces apoptosis in the U‑2OS human osteosarcoma cell line via triggering the mitochondrial pathway

Authors:
- Ya Chen
- Meng Li
- Zhongji Li
- Peng Gao
- Xiao Zhou
- Jianxin Zhang
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Affiliations: Department of Pharmacy, People's Hospital of Zhangqiu, Shandong 250200, P.R. China, Department of Orthopaedic Surgery, People's Hospital of Zhangqiu, Shandong 250200, P.R. China, Spinal Department of Orthopaedic Surgery, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong 250014, P.R. China
Published online on: November 19, 2015 https://doi.org/10.3892/mmr.2015.4583
Pages: 817-822

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Abstract

Bufalin has been shown to induce apoptosis in osteosarcoma cells; however, the underlying mechanism has not been elucidated. The purpose of the present study was to investigate whether mitochondria‑mediated signaling pathways trigger the process of apoptosis in the U‑2OS osteosarcoma cell line. Bufalin inhibited the proliferation and induced apoptosis in U‑2OS cells in a time- and dose‑dependent manner. Bufalin‑induced apoptosis was accompanied with a significant reduction of the mitochondrial membrane potential, release of mitochondrial cytochrome c into the cytosol, activation of caspase‑3, caspase‑9 and poly(adenosine diphosphate ribose) polymerase, as well as downregulation of B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein. Cyclosporin A, a specific inhibitor of the mitochondrial permeability transition pore, attenuated bufalin-induced apoptosis. In conclusion, the present study revealed that bufalin induced apoptosis in the U‑2OS human osteosarcoma cell line via triggering of the mitochondrial pathway.

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1	Mirabello L, Troisi RJ and Savage SA: Osteosarcoma incidence and survival rates from 1973 to 2004: Data from the surveillance, epidemiology and end results program. Cancer. 115:1531–1543. 2009. View Article : Google Scholar : PubMed/NCBI
2	Ottaviani G and Jaffe N: The epidemiology of osteosarcoma. Cancer Treat Res. 152:3–13. 2009. View Article : Google Scholar
3	Fernandez-Pineda I, Bahrami A, Green JF, McGregor LM, Davidoff AM and Sandoval JA: Isolated subcutaneous metastasis of osteosarcoma 5 years after initial diagnosis. J Pediatr Surg. 46:2029–2031. 2011. View Article : Google Scholar : PubMed/NCBI
4	Sampo MM, Tarkkanen M, Kivioja AH, Taskinen MH, Sankila R and Böhling TO: Osteosarcoma in Finland from 1971 through 1990: A nationwide study of epidemiology and outcome. Acta Orthop. 79:861–866. 2008. View Article : Google Scholar : PubMed/NCBI
5	Osborne TS and Khanna C: A review of the association between osteosarcoma metastasis and protein translation. J Comp Pathol. 146:132–142. 2012. View Article : Google Scholar : PubMed/NCBI
6	Bernthal NM, Federman N, Eilber FR, Nelson SD, Eckardt JJ, Eilber FC and Tap WD: Long-term results (>25 years) of a randomized, prospective clinical trial evaluating chemotherapy in patients with high-grade, operable osteosarcoma. Cancer. 118:5888–5893. 2012. View Article : Google Scholar : PubMed/NCBI
7	Jaffe N, Carrasco H, Raymond K, Ayala A and Eftekhari F: Can cure in patients with osteosarcoma be achieved exclusively with chemotherapy and abrogation of surgery? Cancer. 95:2202–2210. 2002. View Article : Google Scholar : PubMed/NCBI
8	Serra M, Scotlandi K, Manara MC, Maurici D, Lollini PL, De Giovanni C, Toffoli G and Baldini N: Establishment and char-acterization of multidrug-resistant human osteosarcoma cell lines. Anticancer Res. 13:323–329. 1993.PubMed/NCBI
9	Li J, Zhang F and Wang S: A polysaccharide from pomegranate peels induces the apoptosis of human osteosarcoma cells via the mitochondrial apoptotic pathway. Tumour Biol. 35:7475–7482. 2014. View Article : Google Scholar : PubMed/NCBI
10	Xie X, Yin J, Jia Q, Wang J, Zou C, Brewer KJ, Colombo C, Wang Y, Huang G and Shen J: Quercetin induces apoptosis in the methotrexate-resistant osteosarcoma cell line U2-OS/MTX300 via mitochondrial dysfunction and dephosphorylation of Akt. Oncol Rep. 26:687–693. 2011.PubMed/NCBI
11	Zhao XH, Xu ZR, Zhang Q and Yang YM: Simvastatin protects human osteosarcoma cells from oxidative stress-induced apoptosis through mitochondrial-mediated signaling. Mol Med Rep. 5:483–488. 2012.
12	Cao Hong, Shibayama-Imazu T, Masuda Y, Shinki T, Nakajo S and Nakaya K: Involvement of Tiam1 in apoptosis induced by bufalin in HeLa cells. Anticancer Res. 27:245–249. 2007.
13	Dong Y, Yin S, Li J, Jiang C, Ye M and Hu H: Bufadienolide compounds sensitize human breast cancer cells to TRAIL-induced apoptosis via inhibition of STAT3/Mcl-1 pathway. Apoptosis. 16:394–403. 2011. View Article : Google Scholar : PubMed/NCBI
14	Gu W, Liu L, Fang FF, Huang F, Cheng BB and Li B: Reversal effect of bufalin on multidrug resistance in human hepatocellular carcinoma BEL-7402/5-FU cells. Oncol Rep. 31:216–222. 2014.
15	Hu F, Han J, Zhai B, Ming X, Zhuang L, Liu Y, Pan S and Liu T: Blocking autophagy enhances the apoptosis effect of bufalin on human hepatocellular carcinoma cells through endoplasmic reticulum stress and JNK activation. Apoptosis. 19:210–223. 2014. View Article : Google Scholar
16	Kurosawa M, Tani Y, Nishimura S, Numazawa S and Yoshida T: Distinct PKC isozymes regulate bufalin-induced differentiation and apoptosis in human monocytic cells. Am J Physiol Cell Physiol. 280:C459–464. 2001.PubMed/NCBI
17	Yin PH, Liu X, Qiu YY, Cai JF, Qin JM, Zhu HR and Li Q: Anti-tumor activity and apoptosis-regulation mechanisms of bufalin in various cancers: New hope for cancer patients. Asian Pac J Cancer Prev. 13:5339–5343. 2012. View Article : Google Scholar
18	Zhu Z, Li E and Liu Y, Gao Y, Sun H, Wang Y, Wang Z, Liu X, Wang Q and Liu Y: Bufalin induces the apoptosis of acute promyelocytic leukemia cells via the downregulation of survivin expression. Acta Haematol. 128:144–150. 2012. View Article : Google Scholar : PubMed/NCBI
19	Zhu Z, Sun H, Ma G, Wang Z, Li E and Liu Y and Liu Y: Bufalin induces lung cancer cell apoptosis via the inhibition of pi3k/akt pathway. Int J Mol Sci. 13:2025–2035. 2012. View Article : Google Scholar : PubMed/NCBI
20	Chang Y, Zhao Y, Zhan H, Wei X, Liu T and Zheng B: Bufalin inhibits the differentiation and proliferation of human osteosarcoma cell line hMG63-derived cancer stem cells. Tumour Biol. 35:1075–1082. 2014. View Article : Google Scholar
21	Wang D and Bi Z: Bufalin inhibited the growth of human osteo-sarcoma MG-63 cells via down-regulation of Bcl-2/Bax and triggering of the mitochondrial pathway. Tumour Biol. 35:4885–4890. 2014. View Article : Google Scholar : PubMed/NCBI
22	Arora S and Tandon S: Achyranthes aspera root extracts induce human colon cancer cell (colo-205) death by triggering the mitochondrial apoptosis pathway and s phase cell cycle arrest. Scientific World Journal. 2014:1296972014. View Article : Google Scholar : PubMed/NCBI
23	Kluck RM, Bossy-Wetzel E, Green DR and Newmeyer DD: The release of cytochrome c from mitochondria: A primary site for Bcl-2 regulation of apoptosis. Science. 275:1132–1136. 1997. View Article : Google Scholar : PubMed/NCBI
24	Li G, Miskimen KL, Wang Z, Xie XY, Brenzovich J, Ryan JJ, Tse W, Moriggl R and Bunting KD: STAT5 requires the N-domain for suppression of miR15/16, induction of bcl-2 and survival signaling in myeloproliferative disease. Blood. 115:1416–1424. 2010. View Article : Google Scholar :
25	Shimizu S, Takehara T, Hikita H, Kodama T, Miyagi T, Hosui A, Tatsumi T, Ishida H, Noda T and Nagano H: The let-7 family of microRNAs inhibits Bcl-xL expression and potentiates sorafenib-induced apoptosis in human hepatocellular carcinoma. J Hepatol. 52:698–704. 2010. View Article : Google Scholar : PubMed/NCBI
26	Zhang LS, Nakaya K, Yoshida T and Kuroiwa Y: Bufalin as a potent inducer of differentiation of human myeloid leukemia cells. Biochemical and biophysical research communications. 178:686–693. 1991. View Article : Google Scholar : PubMed/NCBI
27	Krenn L and Kopp B: Bufadienolides from animal and plant sources. Phytochemistry. 48:1–29. 1998. View Article : Google Scholar : PubMed/NCBI
28	Sun L, Chen T, Wang X, Chen Y and Wei X: Bufalin induces reactive oxygen species dependent bax translocation and apoptosis in astc-a-1 cells. Evid Based Complement Alternat Med. 2011:2490902011. View Article : Google Scholar :
29	Masuda Y, Kawazoe N, Nakajo S, Yoshida T, Kuroiwa Y and Nakaya K: Bufalin induces apoptosis and influences the expression of apoptosis-related genes in human leukemia cells. Leuk Res. 19:549–556. 1995. View Article : Google Scholar : PubMed/NCBI
30	Yu CH, Kan SF, Pu HF, Jea Chien E and Wang PS: Apoptotic signaling in bufalin- and cinobufagin-treated androgen-dependent and -independent human prostate cancer cells. Cancer Sci. 99:2467–2476. 2008. View Article : Google Scholar : PubMed/NCBI
31	Naranmandura H, Chen X, Tanaka M, Wang WW, Rehman K, Xu S, Chen Z, Chen SQ and Suzuki N: Release of apoptotic cytochrome C from mitochondria by dimethylarsinous acid occurs through interaction with voltage-dependent anion channel in vitro. Toxicol Sci. 128:137–146. 2012. View Article : Google Scholar : PubMed/NCBI
32	Ola MS, Nawaz M and Ahsan H: Role of Bcl-2 family proteins and caspases in the regulation of apoptosis. Mol Cell Biochem. 351:41–58. 2011. View Article : Google Scholar : PubMed/NCBI
33	Edlich F, Banerjee S, Suzuki M, Cleland MM, Arnoult D, Wang C, Neutzner A, Tjandra N and Youle RJ: Bcl-x(L) retrotranslocates Bax from the mitochondria into the cytosol. Cell. 145:104–116. 2011. View Article : Google Scholar : PubMed/NCBI
34	Andre N, Roquelaure B and Conrath J: Molecular effects of cyclosporine and oncogenesis: a new model. Medical hypotheses. 63:647–652. 2004. View Article : Google Scholar : PubMed/NCBI