Apoptosis induced by sonodynamic therapy in human osteosarcoma cells in vitro

Liu,Xing; Li,Wei; Geng,Shuo; Meng,Qing‑Gang; Bi,Zheng‑Gang

doi:10.3892/mmr.2015.3479

Apoptosis induced by sonodynamic therapy in human osteosarcoma cells in vitro

Authors:
- Xing Liu
- Wei Li
- Shuo Geng
- Qing‑Gang Meng
- Zheng‑Gang Bi
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Affiliations: Department of Orthopaedic Surgery, The First Afﬁliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Orthopedic Surgery, The First Hospital of Harbin City, Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: March 12, 2015 https://doi.org/10.3892/mmr.2015.3479
Pages: 1183-1188

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Abstract

The aim of the present study was to investigate the potential effect of hematoporphyrin monomethyl ether-sonodynamic therapy (HMME‑SDT) on MG‑63 osteosarcoma cells. The HMME concentration was kept constant at 20 µg/ml and the MG‑63 osteosarcoma cell line was exposed to ultrasound with an intensity of 1.0 W/cm2 for 30 sec. Cell cytotoxicity was quantified using an MTT assay 6 h after HMME‑SDT. The intracellular localization of HMME was imaged using inverted confocal laser scanning microscopy. Apoptosis was investigated using flow cytometry with Annexin V‑fluorescein isothiocyanate and propidium iodine staining. The cytotoxicity of HMME‑mediated sonodynamic action on MG‑63 cells was significantly higher than that of other treatments, including ultrasound alone, HMME alone and sham treatment. Flow cytometry demonstrated that HMME‑SDT action markedly enhanced the apoptotic rate of MG‑63 cells. The mechanisms of apoptosis were analyzed by measuring the protein expression of poly ADP‑ribose polymerase (PARP), cleaved PARP, procaspase‑3, cleaved caspase‑3 and cleaved caspase‑9. The data demonstrated that HMME‑SDT action markedly induced the apoptosis of MG‑63 cells.

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1	Klein MJ and Siegal GP: Osteosarcoma: anatomic and histologic variants. Am J Clin Pathol. 125:555–581. 2006. View Article : Google Scholar : PubMed/NCBI
2	Tan ML, Choong PF and Dass CR: Osteosarcoma: conventional treatment vs. gene therapy. Cancer Biol Ther. 8:106–117. 2009. View Article : Google Scholar
3	Won KY, Lee CH, Kim YW and Park YK: Primary giant-cell-rich osteosarcoma of the urinary bladder: usefulness of osteocalcin and osteonectin immunohistochemical staining and literature review. Pathology. 43:161–164. 2011. View Article : Google Scholar : PubMed/NCBI
4	Shibaguchi H, Tsuru H and Kuroki M: Sonodynamic cancer therapy: a non-invasive and repeatable approach using low-intensity ultrasound with a sonosensitizer. Anticancer Res. 31:2425–2429. 2011.PubMed/NCBI
5	Rosenthal I, Sostaric JZ and Riesz P: Sonodynamic therapy-a review of the synergistic effects of drugs and ultrasound. Ultrason Sonochem. 11:349–363. 2004.PubMed/NCBI
6	Lv Y, Fang M, Zheng J, et al: Low-intensity ultrasound combined with 5-aminolevulinic acid administration in the treatment of human tongue squamous carcinoma. Cell Physiol Biochem. 30:321–333. 2012. View Article : Google Scholar : PubMed/NCBI
7	Barati AH, Mokhtari-Dizaji M, Mozdarani H, Bathaie SZ and Hassan ZM: Treatment of murine tumors using dual-frequency ultrasound in an experimental in vivo model. Ultrasound Med Biol. 35:756–763. 2009. View Article : Google Scholar : PubMed/NCBI
8	Li H, Fan H, Wang Z, Zheng J and Cao W: Potentiation of scutellarin on human tongue carcinoma xenograft by low-intensity ultrasound. PLoS One. 8:e594732013. View Article : Google Scholar : PubMed/NCBI
9	Bai WK, Shen E and Hu B: The induction of the apoptosis of cancer cell by sonodynamic therapy: a review. Chin J Cancer Res. 24:368–373. 2012. View Article : Google Scholar
10	Wang X, Liu Q, Wang Z, et al: Role of autophagy in sonodynamic therapy-induced cytotoxicity in S180 cells. Ultrasound Med Biol. 36:1933–1946. 2010. View Article : Google Scholar : PubMed/NCBI
11	Jaruga E, Salvioli S, Dobrucki J, et al: Apoptosis-like, reversible changes in plasma membrane asymmetry and permeability and transient modifications in mitochondrial membrane potential induced by curcumin in rat thymocytes. FEBS Lett. 433:287–293. 1998. View Article : Google Scholar : PubMed/NCBI
12	Tang W, Liu Q, Zhang J, Cao B, Zhao P and Qin X: In vitro activation of mitochondria-caspase signaling pathway in sonodynamic therapy-induced apoptosis in sarcoma 180 cells. Ultrasonics. 50:567–576. 2010. View Article : Google Scholar : PubMed/NCBI
13	Duvshani-Eshet M, Benny O, Morgenstern A and Machluf M: Therapeutic ultrasound facilitates antiangiogenic gene delivery and inhibits prostate tumor growth. Mol Cancer Ther. 6:2371–2382. 2007. View Article : Google Scholar : PubMed/NCBI
14	Nie F, Xu HX, Lu MD, Wang Y and Tang Q: Anti-angiogenic gene therapy for hepatocellular carcinoma mediated by micro-bubble-enhanced ultrasound exposure: an in vivo experimental study. J Drug Target. 16:389–395. 2008. View Article : Google Scholar : PubMed/NCBI
15	Fuentes-Prior P and Salvesen GS: The protein structures that shape caspase activity, specificity, activation and inhibition. Biochem J. 384:201–232. 2004. View Article : Google Scholar : PubMed/NCBI
16	Ribe EM, Serrano-Saiz E, Akpan N and Troy CM: Mechanisms of neuronal death in disease: defining the models and the players. Biochem J. 415:165–182. 2008. View Article : Google Scholar : PubMed/NCBI
17	Denecker G, Ovaere P, Vandenabeele P and Declercq W: Caspase-14 reveals its secrets. J Cell Biol. 180:451–458. 2008. View Article : Google Scholar : PubMed/NCBI
18	Martinon F and Tschopp J: Inflammatory caspases and inflammasomes: master switches of inflammation. Cell Death Differ. 14:10–22. 2007. View Article : Google Scholar
19	Lamkanfi M, Festjens N, Declercq W, Vanden Berghe T and Vandenabeele P: Caspases in cell survival, proliferation and differentiation. Cell Death Differ. 14:44–55. 2007. View Article : Google Scholar
20	Lipponen P: Apoptosis in breast cancer: relationship with other pathological parameters. Endocr Relat Cancer. 6:13–16. 1999. View Article : Google Scholar
21	Wang P, Xu CS, Xu J, Wang X and Leung AW: Hypocrellin B enhances ultrasound-induced cell death of nasopharyngeal carcinoma cells. Ultrasound Med Biol. 36:336–342. 2010. View Article : Google Scholar
22	Wang P, Xu C, Xia X, et al: Mitochondrial damage in nasopharyngeal carcinoma cells induced by ultrasound radiation in the presence of hypocrellin B. J Ultrasound Med. 29:43–50. 2010.
23	Tang W, Liu Q, Wang X, Wang P, Zhang J and Cao B: Potential mechanism in sonodynamic therapy and focused ultrasound induced apoptosis in sarcoma 180 cells in vitro. Ultrasonics. 49:786–793. 2009. View Article : Google Scholar : PubMed/NCBI
24	He Y, Xia X, Xu C, et al: 5-Aminolaevulinic acid enhances ultrasound-induced mitochondrial damage in K562 cells. Ultrasonics. 50:777–781. 2010. View Article : Google Scholar : PubMed/NCBI
25	Wang XB, Liu QH, Mi N, et al: Sonodynamically induced apoptosis by protoporphyrin IX on hepatoma-22 cells in vitro. Ultrasound Med Biol. 36:667–676. 2010. View Article : Google Scholar : PubMed/NCBI
26	Dai S, Hu S and Wu C: Apoptotic effect of sonodynamic therapy mediated by hematoporphyrin monomethyl ether on C6 glioma cells in vitro. Acta Neurochir (Wien). 151:1655–1661. 2009. View Article : Google Scholar
27	Yumita N, Umemura S, Magario N, Umemura K and Nishigaki R: Membrane lipid peroxidation as a mechanism of sonodynamically induced erythrocyte lysis. Int J Radiat Biol. 69:397–404. 1996. View Article : Google Scholar : PubMed/NCBI
28	Park JW, Ryter SW and Choi AM: Functional significance of apoptosis in chronic obstructive pulmonary disease. COPD. 4:347–353. 2007. View Article : Google Scholar : PubMed/NCBI
29	Schuler M and Green DR: Mechanisms of p53-dependent apoptosis. Biochem Soc Trans. 29:684–688. 2001. View Article : Google Scholar : PubMed/NCBI
30	Haupt S, Berger M, Goldberg Z and Haupt Y: Apoptosis-the p53 network. J Cell Sci. 116:4077–4085. 2003. View Article : Google Scholar : PubMed/NCBI
31	Martin DA and Elkon KB: Mechanisms of apoptosis. Rheum Dis Clin North Am. 30:441–454. 2004. View Article : Google Scholar : PubMed/NCBI
32	Yumita N, Okudaira K, Momose Y and Umemura S: Sonodynamically induced apoptosis and active oxygen generation by gallium-porphyrin complex, ATX-70. Cancer Chemother Pharmacol. 66:1071–1078. 2010. View Article : Google Scholar : PubMed/NCBI
33	Juffermans LJ, Dijkmans PA, Musters RJ, Visser CA and Kamp O: Transient permeabilization of cell membranes by ultrasound-exposed microbubbles is related to formation of hydrogen peroxide. Am J Physiol Heart Circ Physiol. 291:H1595–H1601. 2006. View Article : Google Scholar : PubMed/NCBI
34	Kumon RE, Aehle M, Sabens D, et al: Spatiotemporal effects of sonoporation measured by real-time calcium imaging. Ultrasound Med Biol. 35:49–506. 2009. View Article : Google Scholar
35	Hutcheson JD, Schlicher RK, Hicks HK and Prausnitz MR: Saving cells from ultrasound-induced apoptosis: quantification of cell death and uptake following sonication and effects of targeted calcium chelation. Ultrasound Med Biol. 36:1008–1021. 2010. View Article : Google Scholar : PubMed/NCBI
36	Tabuchi Y, Takasaki I, Zhao QL, et al: Genetic networks responsive to low-intensity pulsed ultrasound in human lymphoma U937 cells. Cancer Lett. 270:2862008. View Article : Google Scholar : PubMed/NCBI