Journal of Prevention and Treatment for Stomatological Diseases ›› 2019, Vol. 27 ›› Issue (10): 621-626.doi: 10.12016/j.issn.2096-1456.2019.10.002

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Effects of X-ray irradiation on proliferation and RANTES expression of the mouse osteogenic precursor cell line MC3T3-E1

LU Weiying1,LIU Ping2,CHEN Jiawei3,LIU Shuying4,XU Pingping5()   

  1. 1. Stomatology Center,Shunde Hospital, Southern Medical University (The First People′s Hospital of Shunde), Foshan 528308, China;
    2. Department of Endodontics,Stomatological Hospital, Southern Medical University,Guangzhou 510280,China
    3. Depertmentt.of Radiotherapy,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,China
    4. Department of Periodontics,Stomatological Hospital,Southern Medical University,Guangzhou 510280,China
    5. Department of Oral and Maxillofacial Surgery, Stomatological Hospital,Southern Medical University,Guangzhou 510280,China;
  • Received:2018-12-30 Revised:2019-05-20 Online:2019-10-20 Published:2019-10-20
  • Contact: Pingping XU E-mail:gdskqyykjk@163.com

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Abstract:

Objective To investigate the effects of X-ray irradiation on the proliferation and protein secretion in vitro in a cultured mouse osteoblast precursor cell line (MC3T3-E1) and to provide a basis for the prevention and treatment of radiation-induced bone injury.Methods Mouse osteoblast precursor cells (MC3T3-E1) in the logarithmic growth phase were divided into a 0 Gy control group, 2 Gy group, 4 Gy group and 8 Gy group according to the irradiation dose. The corresponding dose of X-rays was applied, and the changes in cell morphology after irradiation were observed. Cell proliferation was detected by the CCK-8 method, and alkaline phosphorus was detected at 7 days after irradiation. The alkaline phosphatase (ALP) activity and expression of secreted proteins were detected by high-throughput protein chip technology. The differential expression of proteins due to radiation damage was screened and verified by an ELISA.Results After irradiation, the soma and nucleus of MC3T3-E1 cells were enlarged. Compared to the control group, the cell proliferation rates of the 4 Gy and 8 Gy groups were significantly decreased starting at 3 days in culture (P < 0.05), and the cell proliferation decreased in a dose-dependent manner. After 7 days of culture, the ALP activity in the cells of each irradiated group was lower than that of the control group, and the difference was statistically significant (P < 0.05). Compared to the control group, 36 differentially expressed proteins were found in the supernatant after irradiation. Among them, the content of regulated upon activation, normal T cell expression and secreted factor (RANTES) was significantly increased in a dose-dependent manner.Conclusion A certain dose of irradiation can inhibit the proliferation of mouse osteoblast precursor cells (MC3T3-E1), and RANTES may be an important signaling factor induced by radiation damage.

Key words: irradiation, radioactive osteomyelitis of jaws, osteoblast, MC3T3-E1, cell proliferation, protein chips, secretory protein, regulated upon activation, normal T cell expressed and secreted factor（RANTES）

CLC Number: 

  • R782.3 +2

Figure 1

Cell morphology changes after exposure to different radiation doses × 200"

Table 1

The OD value of cells in a CCK8 assay after exposure to different radiation doses"

培养时间/d 辐照剂量/Gy F P
0 2 4 8
1 0.615 ± 0.037 0.680 ± 0.188 0.453 ± 0.037 0.486 ± 0.057 3.338 0.077
3 1.461 ± 0.108 1.453 ± 0.055 1.231 ± 0.082** 1.221 ± 0.058** 8.646 0.007
5 1.904 ± 0.102 1.841 ± 0.052 1.708 ± 0.036** 1.483 ± 0.014** 28.270 < 0.001
7 2.193 ± 0.214 2.172 ± 0.094 2.136 ± 0.080 1.712 ± 0.049** 9.918 0.005

Figure 2

Comparison of intracellular ALP activity after exposure to different radiation doses"

Figure 3

Expression levels of RANTES in different cell supernatants determined by protein chip technology"

Figure 4

Expression levels of RANTES in different cell supernatants determined by ELISA"

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