ADAM28反义寡核苷酸对人牙龈成纤维细胞内ADAM28的基因表达的影响
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摘要
目的探讨金属蛋白酶解离素28(ADAM28)反义寡核苷酸对人牙龈成纤维细胞(HGFs)内ADAM28表达的抑制作用。方法设计合成与ADAM28 mRNA特异序列互补的AS-ODN和正义对照S-ODN转染HGFs后,通过RT-PCR和Western blot检测细胞中ADAM28的表达差异。结果 ADAM28 AS-ODN转染HGFs后,在转录、翻译和蛋白水平HGFs内ADAM28的表达均明显减弱,统计学分析有显著性差异(P<0.05)。结论 ADAM28 AS-ODN可有效阻断HGFs内ADAM28的基因表达。
Objective To investigate the inhibition of ADAM28 antisense oligodeoxynucleotide(AS-ODN) on the expressions of ADAM28 in human gingival fibroblasts(HGFs). Methods The sequences specifically targeted at ADAM28 mRNA were designed and synthesized as ADAM28 AS-ODN and S-ODN, and ADAM28 AS-ODN and S-ODN were transfected into HGFs. The expression differences of ADAM28 in HGFs were detected by RT-PCR and Western blot assays. Results The expressions of ADAM28 in HGFs were weakened markedly on transcription, translation and protein level after AS-ODN was transfected into HGFs. There was significant difference on statistical analysis(P <0.01).Conclusion ADAM28 AS-ODN can effectively block gene expression of ADAM28 in HGFs.
Objective To investigate the inhibition of ADAM28 antisense oligodeoxynucleotide(AS-ODN) on the expressions of ADAM28 in human gingival fibroblasts(HGFs). Methods The sequences specifically targeted at ADAM28 mRNA were designed and synthesized as ADAM28 AS-ODN and S-ODN, and ADAM28 AS-ODN and S-ODN were transfected into HGFs. The expression differences of ADAM28 in HGFs were detected by RT-PCR and Western blot assays. Results The expressions of ADAM28 in HGFs were weakened markedly on transcription, translation and protein level after AS-ODN was transfected into HGFs. There was significant difference on statistical analysis(P <0.01).Conclusion ADAM28 AS-ODN can effectively block gene expression of ADAM28 in HGFs.
引文
1 Zhao Z,Wen LY,Jin M,et al. ADAM28 participates in the regulation of tooth development. Arch Oral Biol,2006,51(11):996-1005.
2 Seals DF,Courtneidge SA. The ADAMs family of metalloproteases:multidomain proteins with multiple functions. Genes Dev,2003,17(1):7-30.
3 Wolff LF.Guided tissue regeneration in periodontal therapy.Northwest Dent,2000,79(6):23-28,40.
4 蒋少云,陶玉飞,李阳,等.体外诱导人牙龈成纤维细胞多向分化潜能的实验研究.天津医药,2015,43(7):713-716.
5 Chen S,Lyu LX,Huang NP,et al. Inducing the differentiation of human gingival fibroblasts into osteoblasts in vitro. J Med Postgra,2013,26(9):925-927.
6 Zhang QZ,Nquyen AL,Yu WH,et al. Human oral mucosa and gingiva:a unique reservoir for mesenchymal stem cells. J Dent Res,2012,91(11):1011-1018.
7 赵征,魏立,李露嘉,等. ADAM28反义核酸对人牙龈成纤维细胞生物学功能的影响.上海口腔医学, 2014, 23(5):524-530.
8 Zhao Z,Liu H,Wang D. ADAM28 manipulates proliferation,differentiation, and apoptosis of human dental pulp stem cells.J Endod,2011,37(3):332-339.
9 Mitsui Y, Mochizuki S, Kodama T, et al. ADAM28 is overexpressed in human breast carcinomas:Implications for carcinoma cell proliferation through cleavage of insulin-like growth factor binding protein-3. Cancer Res, 2006, 66(20):9913-9920.
10 Zhang XH,Wang CC,Jiang Q,et al. ADAM28 overexpression regulated via the PI3K/AKT pathway is associated with relapse in de novo adult B-cell acute lymphoblastic leukemia. Leuk Res,2015,15:30359-30363.
11 Wood O,Woo J,Seumois G,et al. Gene expression analysis of TIL rich HPV-driven head and neck tumors reveals a distinct B-cell signature when compared to HPV independent tumors. Oncotarget,2016,7(35):56781-56797.
12 Baurakiades E,Costa VH Jr,Raboni SM,et al. The roles of ADAM33, ADAM28, IL-13 and IL-4 in the development of lung injuries in children with lethal non-pandemic acute infectious pneumonia. J Clin Virol,2014,61(4):585-589.
13 Jowett JB,Okada Y,Leedman PJ,et al. ADAM28 is elevated in humans with the metabolic syndrome and is a novel sheddase of human tumour necrosis factor-α. Immunol Cell Biol,2012,90(4):966-973.
14 Herat L,Rudnicka C,Okada Y,et al. The Metalloproteinase ADAM28 Promotes Metabolic Dysfunction in Mice. Int J Mol Sci,2017,18(4):884-894.
15 Mochizuki S, Shimoda M, Shiomi T, et al. ADAM28 is activated by MMP-7(matrilysin-1)and cleaves insulin-like growth factor binding protein-3. Biochem. Biochem Biophys Res Commun,2004,315(1):79-84.
16 Moens TG,Mizielinska S,Niccoli T,et al. Sense and antisense RNA are not toxic in Drosophila models of C9orf72-associated ALS/FTD. Acta Neuropathologica,2018,135(3):445-457.
17 Tufarelli C, Stanley JA, Garrick D, et al. Transcription of antisense RNA leading to gene silencing and methylation as a novel cause of human genetic disease. Nat Genet, 2003, 34(2):157-165.
18 Slavkin HC. Antisense oligonucleotides:An experimental strategy to advance a causal analysis of development. Int J Dev Biol,1995,39:123-126.
19 黎承军,流小舟,施鑫,等. Livin反义寡核苷酸对骨肉瘤MG-63细胞活性的影响.临床肿瘤学杂志, 2017, 22(6):493-499.
20 Niu XY, Peng ZL, Duan WQ, et al. Inhibition of HPV 16E6 oncogene expression by RNA interference in vitro and in vivo. Int J Gynecol Cancer,2006,16(2):743-751.
2 Seals DF,Courtneidge SA. The ADAMs family of metalloproteases:multidomain proteins with multiple functions. Genes Dev,2003,17(1):7-30.
3 Wolff LF.Guided tissue regeneration in periodontal therapy.Northwest Dent,2000,79(6):23-28,40.
4 蒋少云,陶玉飞,李阳,等.体外诱导人牙龈成纤维细胞多向分化潜能的实验研究.天津医药,2015,43(7):713-716.
5 Chen S,Lyu LX,Huang NP,et al. Inducing the differentiation of human gingival fibroblasts into osteoblasts in vitro. J Med Postgra,2013,26(9):925-927.
6 Zhang QZ,Nquyen AL,Yu WH,et al. Human oral mucosa and gingiva:a unique reservoir for mesenchymal stem cells. J Dent Res,2012,91(11):1011-1018.
7 赵征,魏立,李露嘉,等. ADAM28反义核酸对人牙龈成纤维细胞生物学功能的影响.上海口腔医学, 2014, 23(5):524-530.
8 Zhao Z,Liu H,Wang D. ADAM28 manipulates proliferation,differentiation, and apoptosis of human dental pulp stem cells.J Endod,2011,37(3):332-339.
9 Mitsui Y, Mochizuki S, Kodama T, et al. ADAM28 is overexpressed in human breast carcinomas:Implications for carcinoma cell proliferation through cleavage of insulin-like growth factor binding protein-3. Cancer Res, 2006, 66(20):9913-9920.
10 Zhang XH,Wang CC,Jiang Q,et al. ADAM28 overexpression regulated via the PI3K/AKT pathway is associated with relapse in de novo adult B-cell acute lymphoblastic leukemia. Leuk Res,2015,15:30359-30363.
11 Wood O,Woo J,Seumois G,et al. Gene expression analysis of TIL rich HPV-driven head and neck tumors reveals a distinct B-cell signature when compared to HPV independent tumors. Oncotarget,2016,7(35):56781-56797.
12 Baurakiades E,Costa VH Jr,Raboni SM,et al. The roles of ADAM33, ADAM28, IL-13 and IL-4 in the development of lung injuries in children with lethal non-pandemic acute infectious pneumonia. J Clin Virol,2014,61(4):585-589.
13 Jowett JB,Okada Y,Leedman PJ,et al. ADAM28 is elevated in humans with the metabolic syndrome and is a novel sheddase of human tumour necrosis factor-α. Immunol Cell Biol,2012,90(4):966-973.
14 Herat L,Rudnicka C,Okada Y,et al. The Metalloproteinase ADAM28 Promotes Metabolic Dysfunction in Mice. Int J Mol Sci,2017,18(4):884-894.
15 Mochizuki S, Shimoda M, Shiomi T, et al. ADAM28 is activated by MMP-7(matrilysin-1)and cleaves insulin-like growth factor binding protein-3. Biochem. Biochem Biophys Res Commun,2004,315(1):79-84.
16 Moens TG,Mizielinska S,Niccoli T,et al. Sense and antisense RNA are not toxic in Drosophila models of C9orf72-associated ALS/FTD. Acta Neuropathologica,2018,135(3):445-457.
17 Tufarelli C, Stanley JA, Garrick D, et al. Transcription of antisense RNA leading to gene silencing and methylation as a novel cause of human genetic disease. Nat Genet, 2003, 34(2):157-165.
18 Slavkin HC. Antisense oligonucleotides:An experimental strategy to advance a causal analysis of development. Int J Dev Biol,1995,39:123-126.
19 黎承军,流小舟,施鑫,等. Livin反义寡核苷酸对骨肉瘤MG-63细胞活性的影响.临床肿瘤学杂志, 2017, 22(6):493-499.
20 Niu XY, Peng ZL, Duan WQ, et al. Inhibition of HPV 16E6 oncogene expression by RNA interference in vitro and in vivo. Int J Gynecol Cancer,2006,16(2):743-751.