Effect of autophagy on irradiation‑induced damage in osteoblast‑like MC3T3‑E1 cells

Li,Rui; Yang,Wenke; Hu,Xurui; Zhou,Dan; Huang,Ke; Wang,Chenwei; Li,Yi; Liu,Bin

doi:10.3892/mmr.2020.11425

Effect of autophagy on irradiation‑induced damage in osteoblast‑like MC3T3‑E1 cells

Authors:
- Rui Li
- Wenke Yang
- Xurui Hu
- Dan Zhou
- Ke Huang
- Chenwei Wang
- Yi Li
- Bin Liu
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Affiliations: School of Civil Engineering and Mechanics, Lanzhou University, Lanzhou, Gansu 730000, P.R. China, School of Basic Medical Science, Lanzhou University, Lanzhou, Gansu 730000, P.R. China, School/Hospital of Stomatology, Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Department of Cardiology, Gansu Provincial Hospital, Lanzhou, Gansu 730000, P.R. China, School of Civil Engineering and Mechanics, Lanzhou University, Lanzhou, Gansu 730000, P.R. China
Published online on: August 11, 2020 https://doi.org/10.3892/mmr.2020.11425
Pages: 3473-3481
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Autophagy is activated under radiation stress, which serves an important role in maintaining bone homeostasis. However, the underlying mechanisms of irradiation‑induced autophagy in bone homeostasis is not well understood. The present study aimed to determine the effects of radiation‑activated autophagy on pre‑osteoblastic MC3T3‑E1 cells. X‑ray irradiation activated autophagy in a dose‑dependent manner, with an increased fluorescence intensity of monodansylcadaverine staining, increased ratio of microtubule‑associated protein 1 light chain 3β (LC3)‑II/LC3‑I, decreased p62 expression, and increased ATG5 and beclin‑1 expression levels in MC3T3‑E1 cells 72 h after irradiation compared with those in non‑irradiated MC3T3‑E1 cells. Irradiation reduced colony formation and mineralization in a dose‑dependent manner in MC3T3‑E1 cells at 2 and 3 weeks after irradiation, respectively. Decreased levels of alkaline phosphatase activity and runt‑related transcription factor 2 expression were observed at 72 h post‑irradiation. In addition, irradiation‑induced apoptosis was accompanied by a decreased ratio of Bcl‑2/BAX protein and increased the activity of caspase‑3. By contrast, doxycycline (DOX)‑inhibited autophagy attenuated the decreased colony formation and mineralization, and aggravated the increased cell apoptosis in irradiated MC3T3‑E1 cells. Furthermore, the ratio of phosphorylated P38/P38 was observed to be higher following DOX treatment within 1 week of irradiation, which was reversed 2 weeks post‑irradiation. In conclusion, DOX‑inhibited autophagy aggravated X‑ray irradiation‑induced apoptosis at an early stage, but maintained cell proliferation and mineralization at a late stage in irradiated MC3T3‑E1 cells.

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1	Moding EJ, Kastan MB and Kirsch DG: Strategies for optimizing the response of cancer and normal tissues to radiation. Nat Rev Drug Discov. 12:3473–542. 2013. View Article : Google Scholar
2	Cheriex KC, Nijhuis TH and Mureau MA: Osteoradionecrosis of the jaws: A review of conservative and surgical treatment options. J Reconstr Microsurg. 29:69–76. 2013. View Article : Google Scholar : PubMed/NCBI
3	Chronopoulos A, Zarra T, Ehrenfeld M and Otto S: Osteoradionecrosis of the jaws: Definition, epidemiology, staging and clinical and radiological findings. A concise review. Int Dent J. 68:22–30. 2018. View Article : Google Scholar : PubMed/NCBI
4	Rice N, Polyzois I, Ekanayake K, Omer O and Stassen LFA: The management of osteoradionecrosis of the jaws-a review. Surgeon. 13:101–109. 2015. View Article : Google Scholar : PubMed/NCBI
5	Mendenhall WM, Suárez C, Genden EM, de Bree R, Strojan P, Langendijk JA, Mäkitie AA, Smee R, Eisbruch A, Lee AWM, et al: Parameters associated with mandibular osteoradionecrosis. Am J Clin Oncol. 41:1276–1280. 2018. View Article : Google Scholar : PubMed/NCBI
6	Ktistakis NT and Tooze SA: Digesting the expanding mechanisms of autophagy. Trends Cell Biol. 26:624–635. 2016. View Article : Google Scholar : PubMed/NCBI
7	Tang BL: Autophagy in response to environmental stresses: New monitoring perspectives. Ecol Indic. 60:453–459. 2016. View Article : Google Scholar
8	Ko A, Kanehisa A, Martins I, Senovilla L, Chargari C, Dugue D, Mariño G, Kepp O, Michaud M, Perfettini JL, et al: Autophagy inhibition radiosensitizes in vitro, yet reduces radioresponses in vivo due to deficient immunogenic signalling. Cell Death Differ. 21:92–99. 2014. View Article : Google Scholar : PubMed/NCBI
9	Lu C and Xie C: Radiation-induced autophagy promotes esophageal squamous cell carcinoma cell survival via the LKB1 pathway. Oncol Rep. 35:3559–3565. 2016. View Article : Google Scholar : PubMed/NCBI
10	Kim KW, Moretti L, Mitchell LR, Jung DK and Bo L: Combined Bcl-2/mammalian target of rapamycin inhibition leads to enhanced radiosensitization via induction of apoptosis and autophagy in non-small cell lung tumor xenograft model. Clin Cancer Res. 15:6096–6105. 2009. View Article : Google Scholar : PubMed/NCBI
11	Ondrej M, Cechakova L, Durisova K, Pejchal J and Tichy A: To live or let die: Unclear task of autophagy in the radiosensitization battle. Radiother Oncol. 119:265–275. 2016. View Article : Google Scholar : PubMed/NCBI
12	Nollet M, Santucci-Darmanin S, Breuil V, Al-Sahlanee R, Cros C, Topi M, Momier D, Samson M, Pagnotta S, Cailleteau L, et al: Autophagy in osteoblasts is involved in mineralization and bone homeostasis. Autophagy. 10:1965–1977. 2014. View Article : Google Scholar : PubMed/NCBI
13	Liu F, Fang F, Yuan H, Yang D, Chen Y, Williams L, Goldstein SA, Krebsbach PH and Guan JL: Suppression of autophagy by FIP200 deletion leads to osteopenia in mice through the inhibition of osteoblast terminal differentiation. J Bone Miner Res. 28:2414–2430. 2013. View Article : Google Scholar : PubMed/NCBI
14	Han Y, Zhang L, Xing Y, Zhang L, Chen X, Tang P and Chen Z: Autophagy relieves the function inhibition and apoptosis-promoting effects on osteoblast induced by glucocorticoid. Int J Mol Med. 41:800–808. 2018.PubMed/NCBI
15	Li Y, Su J, Sun W, Cai L and Deng Z: AMP-activated protein kinase stimulates osteoblast differentiation and mineralization through autophagy induction. Int J Mol Med. 41:2535–2544. 2018.PubMed/NCBI
16	Hosokawa N, Hara Y and Mizushima N: Generation of cell lines with tetracycline-regulated autophagy and a role for autophagy in controlling cell size. FEBS Lett. 580:2623–2629. 2007. View Article : Google Scholar
17	Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, Øvervatn A, Bjørkøy G and Johansen T: p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem. 282:24131–24145. 2007. View Article : Google Scholar : PubMed/NCBI
18	Li W, Wei S, Liu C, Song M, Wu H and Yang Y: Regulation of the osteogenic and adipogenic differentiation of bone marrow-derived stromal cells by extracellular uridine triphosphate: The role of P2Y2 receptor and ERK1/2 signaling. Int J Mol Med. 37:63–73. 2016. View Article : Google Scholar : PubMed/NCBI
19	Huang S, Wang S, Bian C, Yang Z, Zhou H, Zeng Y, Li H, Han Q and Zhao RC: Upregulation of miR-22 promotes osteogenic differentiation and inhibits adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells by repressing HDAC6 protein expression. Stem Cells Dev. 21:2531–2540. 2012. View Article : Google Scholar : PubMed/NCBI
20	Tian L, He LS, Soni B and Shang HT: Myofibroblasts and their resistance to apoptosis: A possible mechanism of osteoradionecrosis. Clin Cosmet Investig Dent. 4:21–27. 2012. View Article : Google Scholar : PubMed/NCBI
21	Kook SH, Kim KA, Ji H, Lee D and Lee JC: Irradiation inhibits the maturation and mineralization of osteoblasts via the activation of Nrf2/HO-1 pathway. Mol Cell Biochem. 410:255–266. 2015. View Article : Google Scholar : PubMed/NCBI
22	Xu W, Xu L, Chen M, Mao YT, Xie ZG, Wu SL and Dong QR: The effects of low dose x-irradiation on osteoblastic MC3T3-E1 cells in vitro. BMC Musculoskelet Disord. 13:942012. View Article : Google Scholar : PubMed/NCBI
23	Yang B, Tang Q, Post J, Zhou H, Huang XB, Zhang XD, Wang Q, Sun YM and Fan FY: Effect of radiation on the notch signaling pathway in osteoblasts. Int J Mol Med. 31:698–706. 2013. View Article : Google Scholar : PubMed/NCBI
24	Pramojanee SN, Pratchayasakul W, Chattipakorn N and Chattipakorn SC: Low-dose dental irradiation decreases oxidative stress in osteoblastic MC3T3-E1 cells without any changes in cell viability, cellular proliferation and cellular apoptosis. Arch Oral Biol. 57:252–256. 2012. View Article : Google Scholar : PubMed/NCBI
25	Wang C, Blough E, Dai X, Olajide O, Driscoll H, Leidy JW, July M, Triest WE and Wu M: Protective effects of cerium oxide nanoparticles on MC3T3-E1 osteoblastic cells exposed to X-ray irradiation. Cell Physiol Biochem. 38:1510–1519. 2016. View Article : Google Scholar : PubMed/NCBI
26	Szymczyk KH, Shapiro IM and Adams CS: Ionizing radiation sensitizes bone cells to apoptosis. Bone. 34:148–156. 2004. View Article : Google Scholar : PubMed/NCBI
27	Alwood JS, Shahnazari M, Chicana B, Schreurs AS, Kumar A, Bartolini A, Shirazi-Fard Y and Globus RK: Ionizing radiation stimulates expression of pro-osteoclastogenic genes in marrow and skeletal tissue. J Interferon Cytokine Res. 35:480–487. 2015. View Article : Google Scholar : PubMed/NCBI
28	Yang L, Meng H and Yang M: Autophagy protects osteoblasts from advanced glycation end products-induced apoptosis through intracellular reactive oxygen species. J Mol Endocrinol. 56:291–300. 2016. View Article : Google Scholar : PubMed/NCBI
29	Yumoto H, Hirao K, Tominaga T, Bando N, Takahashi K and Matsuo T: Electromagnetic wave irradiation promotes osteoblastic cell proliferation and up-regulates growth factors via activation of the ERK1/2 and p38 MAPK pathways. Cell Physiol Biochem. 35:601–615. 2015. View Article : Google Scholar : PubMed/NCBI
30	Hao Y, Liu C, Huang J, Gu Y, Li H, Yang Z, Liu J, Wang W and Li R: Ghrelin protects against depleted uranium-induced apoptosis of MC3T3-E1 cells through oxidative stress-mediated p38-mitogen-activated protein kinase pathway. Toxicol Appl Pharmacol. 290:116–125. 2016. View Article : Google Scholar : PubMed/NCBI
31	Kralova J, Dvorak M, Koc M and Kral V: p38 MAPK plays an essential role in apoptosis induced by photoactivation of a novel ethylene glycol porphyrin derivative. Oncogene. 27:3010–3020. 2008. View Article : Google Scholar : PubMed/NCBI