锌指蛋白185对胶质瘤细胞增殖影响的研究

The effect of zinc finger protein 185 on the proliferation of human glioma cells

目的：探讨锌指蛋白ZNF185在人脑胶质瘤细胞增殖中的作用。方法标本取自2011年1月—2013年12月于唐山市工人医院就诊，经病理确诊为胶质瘤患者的肿瘤组织，以瘤旁组织作对照。提取不同组织总蛋白， Western-blot检测ZNF185的表达。提取瘤旁组织总RNA，反转录扩增ZNF185编码序列并克隆至pEGFPC2质粒，构建ZNF185表达载体。采用Lipofactamine2000将ZNF185表达载体转染人胶质瘤细胞系SF767，以转染pEGFPC2空载体细胞作为对照。采用流式细胞仪检测细胞周期变化；MTT法检测细胞增殖活性。结果与瘤旁组织相比， ZNF185在人脑胶质瘤组织中表达显著降低（P＜0.01）；转染ZNF185表达载体的胶质瘤细胞与对照细胞相比ZNF185的表达显著增加（P＜0.01）；过表达ZNF185导致SF767细胞G0～G1期细胞比例显著增加（P＜0.05），而S期细胞比例减少（P＜0.05）。同时，过表达ZNF185也导致SF767细胞的增殖速度显著降低（P＜0.05）。结论ZNF185在人脑胶质瘤细胞中发挥抑制细胞增殖的作用。

Objective To explore the role of zinc finger protein （ZNF）185 in the proliferation of human glioma cells. Methods Human glioma tissues and tumor adjacent tissues were obtained from glioma patients diagnosed pathologically in Tangshan Gongren Hospital from January 2011 to December 2013. Total protein was extracted from different tissues. The ZNF185 expression was detected by Western-blot assay. Total RNA was extracted from tumor adjacent tissues. ZNF 185 cod-ing sequence was obtained by RT-PCR and inserted into pEGFPC2 plasmid to construct the ZNF185 expression vector. Li-pofactamine2000 was used to transfect the ZNF185 expression vector to human glioma cell SF767. pEGFPC2 blank vector transfected SF767 cells were used as control. Changes of cell cycle were analyzed by flow cytometry, and cell proliferation was analyzed by MTT assay. Results The expression of ZNF185 decreased significantly in human glioma tissues compared to that of tumor adjacent tissues（P〈0.01）. The over expression of ZNF185 in SF767 resulted in the increased proportion of cell cycle G0-G1phase, but decreased proportion of S phase（P〈0.05）. Furthermore, the cell proliferation was significantly inhibited in the ZNF185 over-expressed SF767 cells compared with that of blank vector transfected cells（P〈0.05）. Con-clusion ZNF185 plays an inhibitory role in cell proliferation of human brain glioma cells.