小鼠钟基因Period2(Per2)抑制乳腺癌细胞生长的研究

目的 研究小鼠节律基因Period2（Per2）对小鼠乳腺癌细胞（EMT6）的生长抑制及诱导凋亡的作用。方法 将pcDNA3．1（＋）-Per2转导入EMT6细胞内，同时设立转染空质粒pcDNA3．1（＋）入细胞的对照组。以RT-PCR、Western blot、流式细胞技术来检测Per2基因和蛋白的表达；采用MTT、细胞生长曲线、细胞平板集落形成实验检测Per2过表达后对小鼠乳腺癌细胞的生长抑制作用；通过流式细胞技术、DNA梯形带、hochst33258荧光染色和电镜技术检测nr2过表达后对小鼠乳腺癌细胞凋亡的影响。结果 小鼠节律基因nr2过表达抑制EMT6细胞的生长和增殖，诱导细胞凋亡率增加。结论 本研究发现小鼠节律基因Per2可能通过使EMT6细胞阻滞于G1期，并诱导细胞凋亡来实现对该肿瘤细胞的生长抑制作用。

Study on Inhibitory Effects of Mouse Circadian Gene Period2(Per2) on EMT6 Cells

Objective To investigate the effects of mouse circadian gene Period2(Per2) overexpression on growth inhibition and apoptosis of EMT6 cells. Method pcDNA3.1( )-Per2 was transfected into EMT6 cells and paralleled with the vector control pcDNA3.1( ). Per2 expression was confirmed by RT-PCR, western blot and flow cytometry. Inhibitory effect of Per2 on cell growth and proliferation were measured by MTT assay, cell count and colony-forming assay. Apoptosis induction of Per2 was detected by flowcytometry, DNA ladder electrophoresis and Hoechst33258 staining and electron microscope. Result The mouse circadian gene Per2 exhibited a statistically significant growth-inhibitory effect and apoptosis-inductory effect on EMT6 cells. Conclusion The results suggest Per2 may inhibit EMT6 cell proliferation by arresting the cells in G1 phase and inducing cellular apoptosis.