辐照对MC3T3-E1细胞TGF-β1和Smads基因及蛋白表达的影响

中国口腔颌面外科杂志 ›› 2015, Vol. 13 ›› Issue (2): 105-109.

辐照对MC3T3-E1细胞TGF-β₁和Smads基因及蛋白表达的影响

刘广龙¹, 刘忠龙¹, 付水霆¹, 胡海生², 何悦¹

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.口腔放疗科, 上海市口腔医学重点实验室,上海 200011

收稿日期:2014-05-26 修回日期:2014-09-11 出版日期:2015-03-20 发布日期:2015-04-23
通讯作者: 何悦,Tel:021-23271699-5656,E-mail:william5218@126.com
作者简介:刘广龙（1987-）,男,硕士研究生,E-mail:lgl515588968@126.com
基金资助:
国家自然科学基金（81271112）

Effects of radiation on the expression of TGF-β₁ and Smads in MC3T3 E1 cells

LIU Guang-long¹, LIU Zhong-long¹, FU Shui-ting¹, HU Hai-sheng², HE Yue¹

1.Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine;
2. Department of Radiotherapy, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology. Shanghai 200011, China

Received:2014-05-26 Revised:2014-09-11 Online:2015-03-20 Published:2015-04-23
Supported by:
Supported by National Natural Science Foundation of China (81271112)

摘要/Abstract

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。

关键词: 放射, MC3T3-E1, TGF-β, 1, Smad3, P-Smad3

Abstract: 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, real-time PCR and Western blot, respectively. Gene and protein expression of TGF-β₁, Smad3 was detected by 2^-ΔΔct and 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.

Key words: Radiation, MC3T3-E1, TGF-β1, Smad3, P-Smad3

中图分类号:

Q691

刘广龙, 刘忠龙, 付水霆, 胡海生, 何悦. 辐照对MC3T3-E1细胞TGF-β₁和Smads基因及蛋白表达的影响[J]. 中国口腔颌面外科杂志, 2015, 13(2): 105-109.

LIU Guang-long, LIU Zhong-long, FU Shui-ting, HU Hai-sheng, HE Yue. Effects of radiation on the expression of TGF-β₁ and Smads in MC3T3 E1 cells[J]. China J Oral Maxillofac Surg, 2015, 13(2): 105-109.

参考文献

[1] Watanabe K, Ikeda K. Osteoblast differentiation and bone formation [J]. Nihon Rinsho, 2009, 67(5): 879-886.
[2] Bongers EM, Haasbeek CJ, Lagerwaard FJ, et al. Incidence and risk factors for chest wall toxicity after risk-adapted stereotactic radiotherapy for early-stage lung cancer [J]. J Thorac Oncol, 2011, 6(12): 2052-2057.
[3] He J, Qiu W, Zhang Z, et al. Effects of irradiation on growth and differentiation-related gene expression in osteoblasts [J]. J Craniofac Surg, 2011, 22(5): 1635-1640.
[4] Czekanska EM, Stoddart MJ, Richards RG, et al. In search of an osteoblast cell model for in vitro research [J]. Eur Cell Mater, 2012, 24(1): 1-17.
[5] Wang D, Christensen K, Chawla K, et al. Isolation and characterization of MC3T3-E1 preosteoblast subclones with distinct in vitro and in vivo differentiation/mineralization potential [J]. J Bone Miner Res, 1999, 14(6): 893-903.
[6] Delanian S, Lefaix JL. The radiation-induced fibroatrophic process: therapeutic perspective via the antioxidant pathway [J]. Radiother Oncol, 2004, 73(2): 119-131.
[7] Darvin P, Joung YH, Yang YM. JAK2-STAT5B pathway and osteoblast differentiation [J]. Jakstat, 2013, 2(4): e24931.
[8] Joung YH, Lim EJ, Darvin P, et al. MSM enhances GH signaling via the Jak2/STAT5b pathway in osteoblast-like cells and osteoblast differentiation through the activation of STAT5b in MSCs [J]. PLoS One, 2012, 7(10): e47477.
[9] Dare A, Hachisu R, Yamaguchi A, et al. Effects of ionizing radiation on proliferation and differentiation of osteoblast-like cells [J]. J Dent Res, 1997, 76(2): 658-664.
[10] Matsumura S, Hiranuma H, Deguchi A, et al. Changes in phenotypic expression of osteoblasts after X irradiation [J]. Radiat Res, 1998, 149(5): 463-471.
[11] Abbas B, Hume SP, Mccullough JS, et al. Early morphological changes in blood capillaries of mouse duodenal villi induced by X-irradiation [J]. J Submicrosc Cytol Pathol, 1990, 22(4): 609-614.
[12] Nicolson GL, Custead SE, Dulski KM, et al. Effects of gamma irradiation on cultured rat and mouse microvessel endothelial cells: metastatic tumor cell adhesion, subendothelial matrix degradation, and secretion of tumor cell growth factors [J]. Clin Exp Metastasis, 1991, 9(5): 457-468.
[13] Lynch SE, Colvin RB, Antoniades HN. Growth factors in wound healing. Single and synergistic effects on partial thickness porcine skin wounds [J]. J Clin Invest, 1989, 84(2): 640-646.
[14] Trojanowska M, Leroy EC, Eckes B, et al. Pathogenesis of fibrosis: type 1 collagen and the skin [J]. J Mol Med (Berl), 1998, 76(3-4): 266-274.
[15] Haase O, Rodemann HP. Fibrosis and cytokine mechanisms: relevant in hadron therapy?[J]. Radiother Oncol,2004,73(Suppl 2): S144-147.
[16] Langdahl BL, Carstens M, Stenkjaer L, et al. Polymorphisms in the transforming growth factor beta 1 gene and osteoporosis [J]. Bone, 2003, 32(3): 297-310.
[17] Sowa H, Kaji H, Yamaguchi T, et al. Smad3 promotes alkaline phosphatase activity and mineralization of osteoblastic MC3T3-E1 cells [J]. J Bone Miner Res, 2002, 17(7): 1190-1199.
[18] Alliston T, Choy L, Ducy P, et al. TGF-beta-induced repression of CBFA1 by Smad3 decreases cbfa1 and osteocalcin expression and inhibits osteoblast differentiation [J]. Embo J, 2001, 20(9): 2254-2272.
[19] Komori T, Yagi H, Nomura S, et al. Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts [J]. Cell, 1997, 89(5): 755-764.
[20] Zúñiga JE, Groppe JC, Cui Y, et al. Assembly of TbetaRI:TbetaRII:TGFbeta ternary complex in vitro with receptor extracellular domains is cooperative and isoform-dependent [J]. J Mol Biol, 2005, 354(5): 1052-1068.
[21] Massagué J, Wotton D. Transcriptional control by the TGF-beta/Smad signaling system [J]. Embo J, 2000, 19(8): 1745-1754.
[22] Annes JP, Munger JS, Rifkin DB. Making sense of latent TGFbeta activation [J]. J Cell Sci, 2003, 116(Pt 2): 217-224.
[23] Frazier WA. Thrombospondins[J]. Curr Opin Cell Biol,1991,3(5): 792-799.
[24] Schultze-Mosgau S, Lehner B, Rödel F, et al. Expression of bone morphogenic protein 2/4, transforming growth factor-beta1, and bone matrix protein expression in healing area between vascular tibia grafts and irradiated bone-experimental model of osteonecrosis [J]. Int J Radiat Oncol Biol Phys, 2005, 61(4): 1189-1196.
[25] Fenner M, Park J, Schulz N, et al. Validation of histologic changes induced by external irradiation in mandibular bone. An experimental animal model[J]. J Craniomaxillofac Surg,2010,38(1): 47-53.
[26] Lau P, Baumstark-khan C, Hellweg CE, et al. X-irradiation-induced cell cycle delay and DNA double-strand breaks in the murine osteoblastic cell line OCT-1 [J]. Radiat Environ Biophys, 2010, 49(2): 271-280.

辐照对MC3T3-E1细胞TGF-β₁和Smads基因及蛋白表达的影响

Effects of radiation on the expression of TGF-β₁ and Smads in MC3T3 E1 cells

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