Abstract：Objective: To evaluate the effect of Notch3 signal pathway on the cell proliferation of thymic epithelial cells for further exploring the mechanisms of thymic atrophy. Methods: Intracellular domain of Notch3(NICD3) DNA was synthesized to construct a lentivirus vector, then it was amplified, digested and sequenced to confirm its sequence. The vector was packaged and used to infect mouse thymic epithelial cells line 1(mTEC1). The fluorescence of the NICD3 was verified and the infection rate was calculated under the fluorescence microscope. Moreover, the cell proliferation was estimated via EdU staining. Results: The NICD3 lentivirus vector was successfully constructed and confirmed by endonuclease digested and sequencing.The infection rate of lentivirus vector was about 20%.The cell proliferation of mTEC1 infected with the lentivirus was inhibited by NICD3(P<0.01). Conclusion: The activation of Notch3 attenuated the mTEC1 proliferation.
［1］HauriHohl MM,Keller MP,Gill J,et al.Donor Tcell alloreactivity against host thymic epithelium limits Tcell development after bone marrow transplantation\[J\].Blood,2007,109(9): 4080-4088.［2］Hince M, Sakkal S, Vlahos K, et al. The role of sex steroids and gonadectomy in the control of thymic involution\[J\]. Cell Immunol,2008,252(1/2):122-138.［3］Shanley DP, Aw D, Manley NR, et al. An evolutionary perspective on the mechanisms of immunosenescence\[J\]. Trends Immunol,2009,30(7):374-381.［4］Beverly LJ, Ascano JM, Capobianco AJ. Expression of JAGGED1 in Tlymphocytes results in thymic involution by inducing apoptosis of thymic stromal epithelial cells\[J\].Genes Immun, 2006,7(6):476-486.［5］Van de Walle I,Waegemans E, De Medts J, et al. Specific Notch receptorligand interactions control human TCRαβ/γδ development by inducing differential Notch signal strength\[J\]. J Exp Med,2013,210(4):683-697.［6］Felli MP, Maroder M, Mitsiadis TA, et al. Expression pattern of notch1, 2 and 3 and Jagged1 and 2 in lymphoid and stromal thymus components: distinct ligandreceptor interactions in intrathymic T cell development\[J\]. Int Immunol,1999,11(7):1017-1025.［7］ArtavanisTsakonas S,Rand MD,Lake RJ.Notch signaling: cell fate control and signal integration in development\[J\]. Science,1999,284(5415):770-776.［8］Suwanjunee S, Wongchana W,Palaga T.Inhibition of gammasecretase affects proliferation of leukemia and hepatoma cell lines through Notch signaling\[J\]. Anticancer Drugs,2008,19(5):477-486.［9］Volanakis EJ, Boothby MR, Sherr CJ. Epigenetic regulation of the Ink4aArf (Cdkn2a) tumor suppressor locus in the initiation and progression of Notch1driven T cell acute lymphoblastic leukemia\[J\]. Exp Hematol, 2013,41(4):377-386.［10］Ferrero I,Koch U,Claudinot S,et al.DL4mediated Notch signaling is required for the development of fetal αβ and γδ T cells\[J\]. Eur J Immunol,2013,43(11):2845-2853.［11］Haji Y,Suzuki M,Moriya K,et al.Activation of Notch1 promotes development of human CD8+ single positive T cells in humanized mice\[J\].Biochem Biophys Res Commun,2014,447(2):346-351.［12］Oh SJ, Ahn S, Jin YH, et al. Notch 1 and Notch 2 synergistically regulate the differentiation and function of invariant NKT cells\[J\]. J Leukoc Biol,2015,98(5):781-789.［13］Yamaguchi E, Chiba S, Kumano K, et al. Expression of Notch ligands, Jagged1, 2 and Delta1 in antigen presenting cells in mice\[J\]. Immunol Lett,2002,81(1):59-64.［14］周晨.树突状细胞诱导胸腺上皮细胞系(MTEC1) 凋亡机制的体外研究\[D\]. 镇江：江苏大学,2015:1-67.［15］Coder BD, Wang H, Ruan L, et al. Thymic involution perturbs negative selection leading to autoreactive T cells that induce chronic inflammation\[J\]. J Immunol, 2015,194(12):5825-5837.［16］Xu L,Zhang Y,Guo R, et al. HES1 promotes extracellular matrix protein expression and inhibits proliferation and migration in human trabecular meshwork cells under oxidative stress\[J\]. Oncotarget,2017,8(13):21818-21833.
2018, Vol. 28 
