辐照对MC3T3-E1细胞TGF-β_1和Smads基因及蛋白表达的影响  

Effects of radiation on the expression of TGF-β_1 and Smads in MC3T3 E1 cells

作　　者：刘广龙[1] 刘忠龙 付水霆 胡海生[2] 何悦[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面-头颈肿瘤科 [2]上海交通大学医学院附属第九人民医院·口腔放疗科,上海市口腔医学重点实验室,上海200011

出　　处：《中国口腔颌面外科杂志》2015年第2期105-109,共5页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81271112)~~

摘　　要：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。PURPOSE： To investigate the effects of radiation on cell proliferation and expression of TGF-β1 and Smad3/P-Smad3 in MC3T3-E1 cells. METHODS： The mouse embryos ossification cell line MC3T3-E1 was used, which was given a single dose of 8 Gy by 4MV linear accelerator. Cell proliferation after irradiation was detected by CCK-8, realtime PCR and Western blot, respectively. Gene and protein expression of TGF-β1, Smad3 was detected by 2-ΔΔctand densitometric analysis were used to analyze the expression level of genes and proteins, respectively. SPSS 13.0 software package was used for statistical analysis. RESULTS： After irradiation of 8 Gy, cell growth slowed significantly compared with the control group; the gene expression of Smad3 and TGF-β1 had no significant change after irradiation for 0.5 h and 1 h（P〉0.05）, until 1.5 h（P〈0.05）; the protein expression of P-Smad3 and TGF-β1 raised obviously on the first day. The difference between exposure group and control group gradually decreased on day 3 and 5. The protein expression of TGF-β1 and P-Smad3 reached peak on the first and third day, respectively. CONCLUSIONS： 8 Gy dose irradiation inhibits cell proliferation and increases the expression of TGF-β1 and Smad3/P-Smad3 on gene and protein level.

关 键 词：放射 MC3T3-E1 TGF-Β1 SMAD3 P-Smad3 

分 类 号：Q274[生物学—细胞生物学] R329.2[医药卫生—人体解剖和组织胚胎学]