Sawajiri Masahiko, Nomura Yuji, Bhawal Ujjal Kumar, Nishikiori Ryo, Okazaki Masayuki, Mizoe Jun'etsu, Tanimoto Keiji
Department of Oral and Maxillofacial Radiology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Bull Exp Biol Med. 2006 Nov;142(5):618-24. doi: 10.1007/s10517-006-0433-4.
We investigated the effects of carbon ion and gamma-irradiation on osteoblastic MC3T3-E1 cells by comparing mRNA expression levels for RANKL and osteoprotegerin by RT-PCR. MC3T3-E1 cells were irradiated with 2, 4, or 6 Gy of carbon ions or gamma-rays, and total RNA was harvested 1, 2, 3, 5, or 7 days after irradiation. The RANKL mRNA/OPG mRNA ratio in carbon ion-irradiated MC3T3-E1 cells was lower, while in gamma-irradiated MC3T3-E1 cells this ratio was higher than in non-irradiated cells. To evaluate osteoclastogenesis of MC3T3-E1 cells, carbon ion- or gamma-irradiated cells were co-cultured with non-irradiated cells from murine bone marrow. Staining for tartrate-resistant acid phosphatase (TRAP) in co-cultures showed that carbon ion irradiation suppressed osteoclastogenesis. This result is consistent with the lower RANKL/OPG mRNA ratio for carbon ion-irradiated cells. These results suggest that carbon ion irradiation acts primarily on osteoblastic cells, leading to a decrease in the RANKL/OPG mRNA ratio. This effect, in turn, leads to a decrease in osteoclastogenesis and osteoclast activity, which results in an increase in bone volume.
我们通过逆转录聚合酶链反应(RT-PCR)比较RANKL和骨保护素的mRNA表达水平,研究了碳离子和γ射线对成骨细胞MC3T3-E1细胞的影响。用2、4或6 Gy的碳离子或γ射线照射MC3T3-E1细胞,并在照射后1、2、3、5或7天收集总RNA。碳离子照射的MC3T3-E1细胞中RANKL mRNA/OPG mRNA比值较低,而γ射线照射的MC3T3-E1细胞中该比值高于未照射细胞。为了评估MC3T3-E1细胞的破骨细胞生成,将碳离子或γ射线照射的细胞与来自小鼠骨髓的未照射细胞共培养。共培养物中抗酒石酸酸性磷酸酶(TRAP)染色显示,碳离子照射抑制了破骨细胞生成。这一结果与碳离子照射细胞中较低的RANKL/OPG mRNA比值一致。这些结果表明,碳离子照射主要作用于成骨细胞,导致RANKL/OPG mRNA比值降低。反过来,这种作用导致破骨细胞生成和破骨细胞活性降低,从而使骨体积增加。