新基因pp3774抑制NIH/3T3细胞生长机制初探

Preliminary study of mechanism on a novel gene pp3774 inhibiting NIH/3T3 cells growth

目的探讨新基因pp3774在NIH／3T3细胞生长调节中的可能机制。方法用pp3774-HA融合表达质粒通过脂质体法转染NIH／3T3细胞,G418筛选,建立稳定细胞系,并用RT-PCR检测pp3774在NIH／3T3细胞中mRNA的表达状况；利用Atlas cDNA array分析pp3774的异位表达对NIH／3T3细胞基因表达谱的影响；用RT-PCR方法验证Atlas cDNA array杂交结果。结果 pp3774超表达可以明显下调cyclin D1、cyclin E、转录因子Dpl等基因的表达,同时可上调P13K p85α、FGF7、MMP-2等基因的表达。上述基因表达的改变可能通过调控细胞周期实现pp3774对NIH／3T3细胞的生长抑制。结论新基因pp3774抑制NIH／3T3细胞生长可能与pp3774基因下调cyclin D1、cyclin E、转录因子Dpl的表达有关。

Objective To explore the possible mechanism of a novel gene pp3774 in NIH/3T3 cell growth regulation. Methods Lipofectamine method was used to transfect pp3774-HA plasmid into NIH/3T3 cell, screened with G418, and stable-expressing cell line was established;RT-PCR was subjected to detect the expression of pp3774 mRNA; Atlas eDNA array was used to analyse the effects of ectopic expression of pp3774 gene on NIH/3T3 genomic profile,which was confirmed with RT-PCR. Results Down-regulation of cyclin D1, cyclin E, transcription factor Dp1 expression, and up-regulation of phosphatidylinositol 3-kinase regulatory subunit, polypeptide 1 （p85a）, growth hormone receptor, fibroblast growth factor 7, matrix metalloproteinase 2 expression were induced by ectopic expression of pp3774. These changes might result in the cell growth inhibition of pp3774 gene in NIH/3T3 cells through cell cycle regulation. Conclusion The inhibition of cell growth in NIH/3T3 may be associated with down-regulation of cyclin D1 ,cyclin E and transcription factor Dp1 by novel gene pp3774.