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目的:探讨放射线对成骨细胞系MC3T3-E1细胞增殖及TGF-β1和Smad3/P-Smad3表达的影响。方法:以小鼠胚胎成骨细胞系MC3T3-E1为研究对象,4MV直线加速器为放射源,给予单一剂量8Gy照射。照射后以CCK-8检测成骨细胞增殖情况,通过荧光定量PCR和Western免疫印迹分别检测TGF-β1、Smad3基因和蛋白的表达,并分别用2-ΔΔct法及光密度分析法分析基因及蛋白的表达水平。采用SPSS 13.0软件包对数据进行统计学分析。结果:单次8Gy剂量照射的细胞组与对照组相比,生长速度明显减慢;Smad3及TGF-β1基因在照射后0.5 h,1 h组与对照组无显著差异(P>0.05),照射后1.5 h组Smad3及TGF-β1基因表达量与对照组存在显著差异(P<0.05);P-Smad3、TGF-β1蛋白表达在第1天明显上调;第3天、第5天照射组与对照组差异逐渐减小。8Gy照射后第1天,P-Smad3蛋白表达量最高,TGF-β1蛋白表达量第3天最高。结论:8Gy照射剂量可抑制MC3T3-E1细胞增殖,短时间内上调TGF-β1和Smad3/P-Smad3在基因及蛋白水平的表达。
Objective: To investigate the effects of radiation on the proliferation and the expression of TGF-β1 and Smad3 / P-Smad3 in osteoblast cell line MC3T3-E1. Methods: The mouse embryonic osteoblastic cell line MC3T3-E1 was used as the research object. The 4MV linear accelerator was used as the radioactive source, and a single dose of 8 Gy was given. The proliferation of osteoblasts was detected by CCK-8 after irradiation. The expression of TGF-β1 and Smad3 genes and proteins were detected by real-time PCR and Western blot respectively. The gene and protein were analyzed by 2-ΔΔct method and optical density analysis The level of expression. Data were statistically analyzed using SPSS 13.0 software package. Results: Compared with the control group, the growth rate of Smad3 and TGF-β1 gene in the group irradiated by 8Gy single dose was significantly lower than that in the control group (P> 0.05) The expression of Smad3 and TGF-β1 in 1.5 h group was significantly different from that in the control group (P <0.05). The expressions of P-Smad3 and TGF-β1 were significantly up-regulated on the 1st day. On the 3rd and 5th days, Differences in the control group gradually decreased. On the first day after 8Gy irradiation, the expression of P-Smad3 protein was the highest, and the expression of TGF-β1 protein was the highest on the third day. CONCLUSION: 8Gy irradiation can inhibit the proliferation of MC3T3-E1 cells and up-regulate the expression of TGF-β1 and Smad3 / P-Smad3 at a gene and protein level in a short time.