CAJ | 학술논문

目的：探索ZNF330对红系分化的影响及作用机制。方法用hemin诱导K562细胞向红系分化，用real-time PCR检测ZNF330的表达；ZNF330的RNAi 慢病毒颗粒感染CD34＋细胞并向红系诱导后用real-time PCR检测CD235a和γ-globin的表达。在293T／17细胞中，用双荧光报告系统检测ZNF330是否具有转录激活作用；在293T／17细胞中，用Co-IP检测ZNF330与mRNA降解途径中ZNF408的相互作用。结果在hemin诱导的K562细胞向红系分化过程中ZNF330表达逐渐升高；抑制ZNF330后，CD34＋细胞中红系分化标志CD235 a和γ-globin的表达下降（ P＜0.05）；未检测到ZNF330的转录激活结构域；双向Co-IP证明ZNF330与ZNF408是相互作用蛋白。结论ZNF330促进红系细胞分化，一个可能的机制是通过与一个参与mRNA降解途径的因子ZNF408的相互作用。
목적：탐색ZNF330대홍계분화적영향급작용궤제。방법용hemin유도K562세포향홍계분화，용real-time PCR검측ZNF330적표체；ZNF330적RNAi 만병독과립감염CD34＋세포병향홍계유도후용real-time PCR검측CD235a화γ-globin적표체。재293T／17세포중，용쌍형광보고계통검측ZNF330시부구유전록격활작용；재293T／17세포중，용Co-IP검측ZNF330여mRNA강해도경중ZNF408적상호작용。결과재hemin유도적K562세포향홍계분화과정중ZNF330표체축점승고；억제ZNF330후，CD34＋세포중홍계분화표지CD235 a화γ-globin적표체하강（ P＜0.05）；미검측도ZNF330적전록격활결구역；쌍향Co-IP증명ZNF330여ZNF408시상호작용단백。결론ZNF330촉진홍계세포분화，일개가능적궤제시통과여일개삼여mRNA강해도경적인자ZNF408적상호작용。
Objective To explore the effects of ZNF330 on erythroid differentiation of K562 cells and underlying mechanism .Methods Realtime PCR was performed to detect the expression of ZNF 330 in K562 cells induced by hemin .After CD34 +cells being infected by the recombination lentivirus ZNF 330-RNAi, Realtime PCR was applied to detect the expression of CD235a and γ-globin.The luciferase report assay was performed to examine if ZNF 330 could act as a trans-acting factor in 293T/17 cells.Co-Immunoprecipitation (Co-IP) was applied in 293T/17 cells to detect the interaction between ZNF 330 and ZNF408 which was involved in mRNA degradation .Results The ex-pression of ZNF330 was up-regulated after hemin treatment .The expression of CD235a andγ-globin decreased after inhibition expression of ZNF 330 had no effect on report gene .Co-Ip in two ways confirmed the direct binding be-tween ZNF330 and ZNF408 .Conclusions ZNF330 can promote erythroid differentiation , and a possible mecha-nism is that ZNF330 inhibits the function of ZNF 408 , a factor that is involved in mRNA degradation , through the interaction between the two proteins .