| Li Yi,Wang Yunlai,Liu Jun,et al.Effects of the overexpression imprinted gene PHLDA2 on radiosensitivity of osteosarcoma[J].Chinese Journal of Radiological Medicine and Protection,2014,34(4):267-270,278 |
| Effects of the overexpression imprinted gene PHLDA2 on radiosensitivity of osteosarcoma |
| Received:October 31, 2013 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.04.007 |
| KeyWords:Osteosarcoma Imprinted gene PHLDA2 Radiosensitivity Cell apoptosis |
| FundProject:国家自然科学基金（81201756）；云南省自然科学基金（2011FZ316，2012FD090） |
| Author Name | Affiliation | E-mail | | Li Yi | Department of Oncology, Kunming General Hospital of People's Liberation Army, Kunming 650032, China | | | Wang Yunlai | 解放军总医院放疗科 | | | Liu Jun | Department of Oncology, Kunming General Hospital of People's Liberation Army, Kunming 650032, China | | | Li Jicong | Department of Oncology, Kunming General Hospital of People's Liberation Army, Kunming 650032, China | | | Chen Hong | Department of Oncology, Kunming General Hospital of People's Liberation Army, Kunming 650032, China | hongchen2000@tom.com |
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| Abstract:: |
| Objective To study the effects of PHLDA2 overexpression on radiosensitivity and the underlying mechanisms in human osteosarcoma U2OS cell line. Methods To obtain the subclone, cells were exposed to G418 persistently after transfection of pEGFP-C3-PHLDA2 vector into U2OS cells. Three groups of blank control(U2OS), negative control(U2OS-neo)and transfected group(U2OS-PHLDA2) were used. The expression of PHLDA2 in the subclone cells was determined by Western blot. After exposure to X-ray irradiation, cellular growth activity and survival were detected by CKK-8 assay and colony formation assay, respectively. The cell apoptosis was measured by the Annexin V/PI staining, and the apoptotic protein was analyzed by Western blot. The in-vivo effects of PHLDA2 on irradiation were evaluated by xenografts. Results Compared with U2OS group and U2OS-neogroup, the subclone cells were successfully obtained by G418 selection, in which the expression of PHLDA2 was upregulated(t=13.73,16.28, P<0.05). In vitro, PHLDA2 overexpression significantly enhanced the response to radiation in U2OS cells with a reduction of colony survival and proliferation with the increase of doses (t=5.00-8.23,P<0.05;t=－2.52—1.26, P<0.05). In vivo, PHLDA2-upregulated xenografts had more radiosensitivity than control groups with a significant inhibition of tumor growth (t=3.27,2.91,P<0.05). After 8 Gy irradiation, the apoptosis was significantly increased(t=10.11,9.61,P<0.05), accompanied with the activation of Caspased-3 in U2OS-PHLDA2 cells, which was presented by upregulation of cleaved Caspase-3 (t=11.26,10.72,P<0.05). Conclusions Exogenetic expression of PHLDA2 could significantly enhance the radiosensitivity of human osteosarcoma cells, which may be attributed to the activation of Caspase-3 that increases irradiation-induced apoptosis. |
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