【摘要】 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、Westernblot、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Per2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Per2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。更多还原

【Abstract】 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.更多还原