National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, PR China.
Mol Med Rep. 2011 Nov-Dec;4(6):1131-7. doi: 10.3892/mmr.2011.564. Epub 2011 Aug 22.
Previously, we characterized a zinc finger protein gene HZF1 (ZNF16) and demonstrated that it played a significant role in the erythroid and megakaryocytic differentiation of K562 cells by knockdown of the gene. In this study, we examined the effect of HZF1 on the proliferation and apoptosis of K562 cells and identified the possible mechanism for this effect. By lentivirus-mediated gene transfer, we obtained stable K562 transductants with HZF1 overexpression (K562/WPXL-HZF1) and stable control transductants (K562/WPXL). Significantly rapid cell amplification was observed in K562/WPXL-HZF1 cells compared to K562/WPXL cells. The cell cycles of the two transductants were analyzed and the results demonstrated that HZF1 overexpression promoted the S to G2/M phase transition. Additionally, we found that the overexpression of HZF1 slightly inhibits the apoptosis of K562 cells induced by sodium arsenate. Furthermore, using a yeast two-hybrid (Y2H) system we identified the HZF1-interacting proteins and screened 29 potential binding partners of HZF1. Using a co-immunoprecipitation (Co-IP) assay, we confirmed the interaction between HZF1 and the inhibitor of cyclin-dependent kinase (CDK) interacting with cyclin A1 (INCA1), and proved that this interaction leads to the inhibition of INCA1 function, which rescued the activity of CDK2 inhibited by INCA1. In conclusion, our results identified novel functions of the HZF1 gene and revealed a possible mechanism through which HZF1 affects K562 cell proliferation.
先前，我们鉴定了一个锌指蛋白基因 HZF1（ZNF16），并通过基因敲低证明其在 K562 细胞的红系和巨核细胞分化中发挥重要作用。在本研究中，我们研究了 HZF1 对 K562 细胞增殖和凋亡的影响，并确定了这种影响的可能机制。通过慢病毒介导的基因转移，我们获得了 HZF1 过表达（K562/WPXL-HZF1）和稳定对照转导物（K562/WPXL）的稳定 K562 转导物。与 K562/WPXL 细胞相比，K562/WPXL-HZF1 细胞中观察到明显快速的细胞扩增。分析了两种转导物的细胞周期，结果表明 HZF1 过表达促进了 S 期到 G2/M 期的转变。此外，我们发现 HZF1 的过表达略微抑制了砷酸钠诱导的 K562 细胞凋亡。此外，我们使用酵母双杂交（Y2H）系统鉴定了 HZF1 的相互作用蛋白，并筛选出 29 个 HZF1 的潜在结合伙伴。通过共免疫沉淀（Co-IP）实验，我们证实了 HZF1 与细胞周期蛋白依赖性激酶（CDK）相互作用的抑制剂（INCA1）之间的相互作用，并证明这种相互作用导致 INCA1 功能受到抑制，从而挽救了 INCA1 抑制的 CDK2 的活性。总之，我们的结果确定了 HZF1 基因的新功能，并揭示了 HZF1 影响 K562 细胞增殖的可能机制。
