Notch3胞内活化片段抑制小鼠胸腺上皮细胞系增殖

目的: 研究Notch3信号对胸腺上皮细胞（thymic epithelial cells，TECs）增殖的影响。方法: 人工合成Notch3胞内活化片段（NICD3）序列，构建慢病毒载体，并进行酶切和测序验证。将NICD3包装后感染小鼠TECs系mTEC1，荧光显微镜观察记录绿色荧光细胞的数量与可见光下细胞数量并计算感染效率。采用EdU增殖试剂盒检测mTEC1细胞增殖率。结果： 酶切和测序证实成功构建NICD3慢病毒表达载体，荧光显微镜观察表明该质粒能较好地感染mTEC1细胞，感染效率约为20%。EdU增殖实验显示过表达NICD3可抑制mTEC1细胞增殖（P<0.01）。 结论： Notch3信号抑制mTEC1细胞的增殖。

Over-expressed NICD3 inhibits the proliferation of mouse thymic epithelial cells line

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.