CAJ | 학술논문

目的：在慢病毒载体pSIH‐H1构建干扰人Notch3基因的载体，在293T细胞包装并在鼻咽癌细胞中检测Notch3的表达。方法沉默Notch3的寡核苷酸序列连接慢病毒载体pSIH‐H1，测序。在293T 细胞包装病毒，测定其滴度。在鼻咽癌细胞中CNE2感染慢病毒，Western Blot分析沉默效果。结果慢病毒载体pSIH‐H1‐Notch3测序正确，在293T细胞中可以观察到GFP的表达，Western Blot证实可以在鼻咽癌细胞中沉默Notch3的表达。结论在慢病毒干扰载体pSIH‐H1构建沉默Notch3载体正确，包装病毒可在鼻咽癌细胞中沉默Notch3，可以用于鼻咽癌干细胞的研究。
목적：재만병독재체pSIH‐H1구건간우인Notch3기인적재체，재293T세포포장병재비인암세포중검측Notch3적표체。방법침묵Notch3적과핵감산서렬련접만병독재체pSIH‐H1，측서。재293T 세포포장병독，측정기적도。재비인암세포중CNE2감염만병독，Western Blot분석침묵효과。결과만병독재체pSIH‐H1‐Notch3측서정학，재293T세포중가이관찰도GFP적표체，Western Blot증실가이재비인암세포중침묵Notch3적표체。결론재만병독간우재체pSIH‐H1구건침묵Notch3재체정학，포장병독가재비인암세포중침묵Notch3，가이용우비인암간세포적연구。
Objective To constrcture Notch3‐silencing lentivirus vector and detect its silencing effect in nasopha‐ryngeal carcinoma .Method The oligonucleotide of silencing Notch3 was ligated with lentivirus vector pSIH‐H1 .The lentivirus expression vector was cotransfected into 293T cells with FIV34N and pMD2G and titre were measured . The effect of silencing Notch3 in nasopharyngeal carcinoma CNE2 were detected by western blot (WB) test .Result The result of vectors sequencing was shown that the silencing lentivirus vector was constructed successfully .After transfection of silencing lentivirus vector ,we observed it GFP‐positive in 293T cells .The effect of silencing Notch3 in nasopharyngeal carcinoma CNE2 were confirmed by WB test .Conclusion The lentivirus vector of silencing Notch3 is constructed successfully and can be used in nasopharyngeal carcinoma sten cell research .