过表达DEPDC1对鼻咽癌细胞CNE-1增殖影响的研究

Effect of overexpression of DEPDC1 on proliferation of nasopharyngeal carcinoma cell line CNE-1

目的:DEPDC1(DEP domain containing 1)是近年来发现的一个肿瘤相关基因,前期研究结果显示其对鼻咽癌发生发展具有重要作用,本研究旨在进一步探讨过表达DEPDC1对鼻咽癌细胞增殖的影响。方法:在鼻咽癌细胞系CNE-1中,分别转染过表达DEPDC1-V1及DEPDC1-V2两种剪接体质粒,瞬时表达48 h后综合利用CCK-8、流式细胞技术及p HH3标记法检测细胞增殖情况。结果:Western blot检测显示在CNE-1细胞瞬时转染DEPDC1-V1和DEPDC1-V2质粒可过表达DEPDC1。CCK-8实验结果表明,过表达DEPDC1-V1及DEPDC1-V2均显著促进CNE-1细胞增殖。流式细胞检测结果表明,与对照组相比过表达DEPDC1-V1或DEPDC1-V2后均导致G2/M期细胞增多。p HH3检测结果显示,过表达DEPDC1-V1或DEPDC1-V2后有丝分裂活跃,细胞周期进程加快。进一步定量RT-PCR检测发现,过表达DEPDC1-V1或DEPDC1-V2导致细胞周期进程关键调节因子Fox M1及其下游靶基因CCNB2、PLK1和MYBL2表达量上调。结论:本研究通过在鼻咽癌CNE-1细胞株中过表达两个剪接体DEPDC1-V1及DEPDC1-V2,证明DEPDC1在鼻咽癌细胞中高表达对促进细胞的增殖具有重要作用。

Objective：DEP domain containing 1（DEPDC1）is a tumor-related gene found in recent years. Previous studies have shown that DEPDC1 plays an important role in the development and progression of nasopharyngeal carcinoma. In this study,DEPDC1 was used to further investigate the effect of DEPDC1 on the proliferation of NPC cells. Methods：In nasopharyngeal carcinoma cell line CNE-1,the two spliceosomes overexpressing DEPDC1-V1 and DEPDC1-V2 were transfected respectively,and transiently expressed for 48 h,then CCK-8,flow cytometry and p HH3 labeling assays were used to detect cell proliferation. Results：Western blot showed that transient transfection of DEPDC1-V1 and DEPDC1-V2 in CNE-1 cells could overexpress DEPDC1. CCK-8 experimental results showed that,over-expression of DEPDC1-V1 and DEPDC1-V2 significantly promoted CNE-1 cell proliferation. Flow cytometry results showed that compared with the control group,overexpression of DEPDC1-V1 or DEPDC1-V2 resulted in an increase of G2/M phase cells. The result of p HH3 labeling assay showed that mitosis was active after DEPDC1-V1 or DEPDC1-V2 being overexpressed,and the cell cycle progression was accelerated. Further quantitative RT-PCR analysis showed that overexpression of DEPDC1-V1 or DEPDC1-V2 led to the up-regulation of Fox M1 and its downstream target genes CCNB2,PLK1 and MYBL2 in cell cycle progression. Conclusion：In this study,overexpression of two spliceosomes,DEPDC1-V1 and DEPDC1-V2,in nasopharyngeal carcinoma CNE-1 cell lines demonstrate that DEPDC1 overexpression in nasopharyngeal carcinoma cells plays an important role in promoting cell proliferation.