生长抑制因子4基因表达对K562细胞生长的影响  

Ad-ING4 inhibits K562 cell growth

作　　者：郁心[1] 张海峰[1] 王金志[1] 谢宇锋[1] 杨吉成[1] 缪竞诚[1] 

机构地区：[1]苏州大学医学院细胞与分子生物学教研室,215123

出　　处：《中华血液学杂志》2007年第6期396-400,共5页Chinese Journal of Hematology

摘　　要：目的研究生长抑制因子4(ING4)基因表达对 K562细胞增殖的影响。方法将经过定点突变技术人源化改造的 ING4(鼠→人)生长抑制因子4基因酶切并连接到 pAdTrack-CMV 转移质粒上,然后与腺病毒质粒 pAdEasy-1共转化 BJ5183细菌,获得重组腺病毒质粒 pAdEasy-1-pAdTrack-CMV-hING4,将它转染 QBI-293A 细胞后收获重组腺病毒 Ad-hING4(以下简称为 Ad-ING4)。将 Ad-ING4感染 K562细胞,光学显微镜下观察病毒感染前后细胞形态的变化,免疫细胞化学分析凋亡因子的改变,激光扫描共聚焦显微镜观察 K562细胞的凋亡,流式细胞术(FCM)检测细胞凋亡率。结果成功构建了人 ING4腺病毒质粒,获得高滴度的重组腺病毒 Ad-ING4,用它感染 K562细胞72 h后,FCM 法测定细胞凋亡率为19.7%,与空病毒对照组相比差异有统计学意义(P<0.01),ING4基因能下调 K562细胞 bcl-2的表达并上调 bax 的表达,多种方法检测结果表明 ING4基因能抑制 K562细胞生长,诱导凋亡。结论重组腺病毒 Ad-ING4可以显著抑制 K562细胞的生长。Objective To observe the effect of recombinant adenovirus Ad-ING4 on K562 cells. Methods Human ING4 recombinant transfer vector pAdTrack-CMV-ING4 was constructed by enzyme digest and ligation of human ING4 gene which was obtained through site specific point mutation of mouse INC,4. The vector was co-transduced into BJ5183 E. coli with pAdEasy-1. The new recombinant adenovirus vector pAdEasy-l-pAdTrack-CMV-hING4 was transfected into QBI-293A cells. To obtain the ING4 recombined adenovirus（Ad-ING4）. Ad-ING4 was used to infect K562 cells. The effect on K562 cells of ING4 was tested by LSCM FCM and immunohistochemistry. Results Human ING4 recombinant adenovirus vector was constructed successfully, and high titre ING4 recombinant adenovirus（Ad-ING4） was obtained. ING4 can down-regulate the expression of bcl-2 and up-regulate expression of bax. The apoptosis of K562 cells induced by ING4 was proved by LSCM FCM and immunohistochemistry. The apoptosis rate was 19.7% （ after 72h ）, which displayed significant difference compared with that of control groups （ P 〈 0.01 ）. Conclusion Ad-ING4 can inhibit the growth of K562 cells and induce the cells apoptosis. The human ING4 recombinant adenoviral vector constructed might provide an approach to the target therapy of tumors.

关 键 词：基因 ING4 细胞系 K562 细胞凋亡 

分 类 号：R363[医药卫生—病理学]