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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。
Objective To study the expression of IER3IP1 gene in the process of differentiation of K562 cells induced by matrine, so as to clarify the dose-effect relationship and time-effect relationship of IER3IP1 gene expression and matrine, and to explore the function of this gene in K562 cells. Methods Trypan blue staining was used to analyze the growth inhibitory effect of matrine on K562 cells; RT-PCR semi-quantitative method was used to observe the expression of IER3IP1 gene in K562 cells treated with matrine at different times and different doses; Stained K562 cells (K562/eYFP-IER3IP1) were used for observation of cell morphology and cell proliferation changes, while cell cycle detection and ultrastructure observation were performed. Results Matrine inhibited the proliferation of K562 cells, and the expression of IER3IP1 gene increased 3 to 4 folds after matrine exposure for 3 hours in a dose-dependent manner. The expression decreased after 6 to 48 hours, which was lower than that of non-bitter. Alkaline treatment group. The proliferation rate of K562/eYFP-IER3IP1 cells was significantly slowed down, and the number of G_0/G_1 phase cells was increased (P<0.05). After 24 h of matrine exposure, erythroid differentiation cells were observed under light microscope and electron microscope. Conclusion Matrine can inhibit the growth of K562 cells and increase the expression of IER3IP1 gene in a dose-dependent manner. The sensitivity of K562 cells transfected with IER3IP1 gene to matrine is increased, suggesting that this gene may be involved in the matrine action. The early response of K562 cells, and play a role in the subsequent erythroid differentiation.