靶向调控TOX3基因对乳腺癌细胞增殖及周期的影响

Effects of Targeted Regulation of TOX3 Gene on Proliferation and Cycle of Human Breast Cancer Cells

目的研究TOX3基因对人乳腺癌细胞增殖能力及细胞周期的影响。方法采用Western blot法检测已构建的稳定沉默TOX3的ZR-75-1细胞和稳定过表达TOX3的MDA-MB-231细胞中TOX3蛋白的表达。MTT法检测过表达和干扰TOX3基因对乳腺癌细胞增殖活性的影响。平板单克隆实验检测过表达TOX3对MDA-MB-231细胞单克隆形成能力的影响,流式细胞术检测沉默或过表达TOX3后ZR-75-1和MDA-MB-231细胞周期的变化。结果与阴性对照组比较,稳定沉默TOX3的乳腺癌ZR-75-1细胞TOX3蛋白表达明显下降,过表达TOX3的MDA-MB-231细胞TOX3蛋白表达明显上调。过表达TOX3后,MDA-MB-231细胞的增殖活性明显增强,单克隆形成能力增强,G 0/G 1期细胞所占比例明显减少,G 2/M期细胞比例增多。干扰TOX3后,ZR-75-1细胞增殖活性下降,G 0/G 1期细胞比例增加,G 2/M期细胞比例减少。结论靶向调控TOX3基因的表达可改变乳腺癌细胞增殖和单克隆形成能力,影响乳腺癌细胞周期。

Objective To investigate the effect of TOX3 gene on proliferation and cycle of human breast cancer cells.Methods Western blot was used to test the expression of TOX3 in ZR-75-1 cells with stably silenced TOX3 gene and MDA-MB-231 cells with stably over-expressed TOX3 gene.The effect of TOX3 on pro1liferation of human breast cancer cells was measured by MTT assay and the effect of over-expressed TOX3 gene on the ability of monoclonal formation of the MDA-MB-231cells was detected by plate colony formation experiment.The flow cytometry was employed to detect the effect of TOX3 on cycle of ZR-75-1 cells and MDA-MB-231 cells.Results The expression of TOX3 protein in ZR-75-1 cells with stably silenced TOX3 gene significantly reduced and up-regulated in MDA-MB-231 cells with stably over-expressed TOX3 gene compared with the negative control group.The over-expression of TOX3 enhanced proliferation activity and the ability of monoclonal formation of the MDA-MB-231cells.The rate of cells in G 0/G 1 phase was reduced and the rate of cells in G 2/M phase was increased significantly.After the silence of TOX3 gene,the proliferation activity of ZR-75-1 cells was declined.The rate of cells in G 0/G 1 phase was increased and the rate of cells in G 2/M phase was decreased.Conclusion Targeted regulation of TOX3 gene may change the ability of proliferation and monoclonal formation of breast cancer cells,affect the cycle of breast cancer cells.