IRE1α重组腺病毒载体对ATDC5软骨干细胞增殖及凋亡的影响.PDFVIP

Med J Chin PLA, Vol. 38, No. 8, August 1, 2013 　639 论　著 IRE 1α重组腺病毒载体对ATD C5软骨干细胞增殖及凋亡的 影响 李祥柱，张鹏，韩晓凤，李美玲，夏飞，郭风劲 TM [摘要]　目的　构建重组人IRE 1α腺病毒，观察其对软骨干细胞ATDC5增殖与凋亡的影响。方法　应用pA dEasy 腺病毒载体系统构建包含全长IRE 1α基因和RNase+Kinase核心结构域的重组穿梭质粒pA dTrack-IRE 1α、pA dTrack-R+K ， 通过电转法分别与腺病毒骨架质粒pA dEasy- 1进行同源重组，获得重组腺病毒pA d-IRE 1α、pA d-R+K 。随后在HEK-293 细胞中包装并扩增重组腺病毒颗粒A d-IRE 1α、A d-R+K 。用直接P CR法鉴定重组腺病毒，后者体外感染软骨干细胞 ATD C5 ，荧光显微镜下观察绿色荧光蛋白(GFP)感染效率，并应用流式细胞仪分别检测在内质网应激(ER S)和非ER S 状态时重组腺病毒对ATD C5细胞增殖和凋亡的影响。结果　酶切鉴定和P CR证实成功构建了重组腺病毒质粒pA d- IRE 1α、pA d-R+K 。流式细胞仪检测结果显示，在ERS状态时，重组腺病毒质粒A d-IRE 1α、A d-R+K感染后ATD C5细胞 G1期比例明显降低，S期细胞比例明显升高(P 0.05) ，细胞凋亡率明显增高(P 0.05) 。结论　含IRE 1α基因的重组腺病 毒感染可促进ATDC5细胞的增殖和凋亡。 [关键词]　IRE 1α；腺病毒科；ATDC5细胞；细胞增殖；细胞凋亡 [中图分类号]　R329.2 [文献标志码]　A [文章编号]　0577-7402 (20 13)08-0639-05 Effects of recombinant adenoviral vector containing IRE1α gene on proliferation and apoptosis of ATDC5 stem cells * LI Xiang-zhu, ZHANG Peng, HAN Xiao-feng, LI Mei-ling, XIA Fei, GUO Feng-jin Department of Cell Biology and Genetics, Chongqing Medical University, Chongqing 400016, China * Corresponding author, E-mail: guo.fengjin@ This work was supported by the National Natural Science Foundation of China , the New Century Excellent Talent Support Project of Education Ministry of China (NCET-12-1090), and the University Excellent Talent Support Project of CQEC (2011-65) [Abstract]　Objective　To construct the recombinant adenoviral vector containing human IRE 1α (type I transmembrane protein kinase/endoribonuclease) gene, and investigate its effects on proliferation and apoptosis o