苦参碱处理的K562细胞中IER3IP1基因表达及其对细胞增殖的影响  

The expression of IER3IP1 gene in K562 cells treated by matrine and its effect on the cell growth

作　　者：王伟佳[1] 张彦[1] 黄凤霞[1] 

机构地区：[1]重庆医科大学临床生化教研室临床检验诊断学省部共建重点实验室,400016

出　　处：《中华血液学杂志》2007年第12期823-827,共5页Chinese Journal of Hematology

基　　金：国家自然科学基金(30171150)

摘　　要：目的研究苦参碱诱导 K562细胞分化过程中 IER3IP1基因的表达,以明确 IER3IP1基因表达与苦参碱作用的量效及时效关系,并初步探讨该基因在 K562细胞中的功能。方法锥虫蓝染色分析苦参碱对 K562细胞的生长抑制作用;用 RT-PCR 半定量方法观察 K562细胞在不同时间、不同剂量苦参碱作用下 IER3IP1基因的表达情况;对 IER3IP1基因重组质粒转染的 K562细胞(K562/eYFP-IER3IP1)作细胞形态学及细胞增殖变化的观察,同时进行细胞周期检测以及超微结构观察。结果苦参碱对 K562细胞有增殖抑制作用,在苦参碱作用3 h 后 IER3IP1基因表达可升高3～4倍,并呈剂量依赖性,其后6～48 h 表达下降,低于非苦参碱处理组。K562/eYFP-IER3IP1细胞的增殖速度显著减缓,G_0/G_1期细胞数升高(P<0.05);且苦参碱作用24 h 后在光镜和电镜下均可见红系分化细胞增多。结论苦参碱抑制 K562细胞生长,同时使 IER3IP1基因表达以剂量依赖的方式瞬时升高,转染了 IER3IP1基因的 K562细胞对苦参碱敏感性增高,提示该基因可能参与了苦参碱作用于 K562细胞的早期反应,并在后续的红系分化中发挥作用。Objectives To investigate the expression status of IER3IP1 gene during matrine induced K562 cell differentiation, and to figure out the function of IER3IP1 gene in K562 cell line. Methods Trypan-blue staining was used to analyze the growth inhibitory effect of matrine on K562 cells. Semi-quantitative RT-PCR was employed to investigate the expression status of IER3IP1 gene treated with different time and dosage of matrine. The alteration of cellular morphology, cellular proliferation and uhra-microstructure were observed and the cell cycle was detected on the recombinant IER3IP1 gene eukaryotic expression vector eYFP- IER3IP1 plasmid transfected K562 cells （ K562/eYFP-IER3IP1 ）. Results Matrine inhibited the growth of K562 cellls and reduced the expression of IER3IP1 gene. The expression level of IER3IP1 gene was transiently increased to three-to-four times in a dose-dependent manner after treated with matrine for 2 - 3 hours. Then, in 6 - 48 hours it maintained at a low level as compared with the control group. The proliferation rate of the K562/eYFP-IER3IP1 cells significantly slowed down with more cells blocked in G0 - G1 phase （ P 〈 0.05 ）. The number of erythroid blast cells began to increase after 24 hours of matrine treatment. At the same time, differentiated erythroid cells could be observed. Conclusions Matrine can inhibit the growth of K562 cells, and transiently increase the expression level of IER3IP1 gene in a dose-dependent manner. The sensitivity of K562 cells to matrine maybe increased after being transfected by the eYFP-IER3IP1 plasmid, indicating a possible involvement of the IER3IP1 gene in the early response of the cells to matrine and its possible role in the erythroid cell differentiation.

关 键 词：基因 IER3IP1 苦参碱 K562细胞 细胞分化 

分 类 号：R285.5[医药卫生—中药学]