敲低SOX2基因抑制乳腺癌干细胞的含量  

Knockdown of SOX2 Gene Inhibits Breast Cancer Stem Cells

作　　者：姜丽娜[1] 程龙[2] 叶棋浓[2] 吕朝晖[1] 

机构地区：[1]解放军总医院内分泌科,北京100853 [2]军事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2016年第1期58-62,共5页Letters in Biotechnology

基　　金：国家自然科学基金(81372161)

摘　　要：目的:构建敲低SOX2基因的慢病毒表达载体,研究敲低SOX2基因对乳腺癌干细胞含量的影响。方法:将SOX2基因短发夹RNA(sh RNA)序列构建到慢病毒表达载体,利用293T细胞包装并获得敲低SOX2基因的病毒,后在MCF-7、T47D细胞系中构建敲低SOX2基因的稳定克隆,Western印迹检测SOX2蛋白的表达水平,q RT-PCR检测SOX2 m RNA水平的变化,利用流式细胞术检测敲低SOX2基因后乳腺癌干细胞含量的变化。结果:构建了表达SOX2基因sh RNA的慢病毒表达载体,在乳腺癌细胞中敲低SOX2基因后,CD44+/CD24-/low及乙醛脱氢酶阳性(ALDH+)细胞的比例明显降低。结论:敲低SOX2基因抑制了乳腺癌干细胞的含量。Objective： To construct the lentiviral vector of RNA interference for SOX2 gene, and to detect its ef- fect on breast cancer stem cells. Methods： The SOX2 short hairpin RNA（shRNA） sequence was constructed to the expression vector. Recombinant lentivirus carrying SOX2 shRNA was packaged and obtained in 293T cells. MCF-7 cells and T47D cells were infected with the lentivirus and selected for stable cells. SOX2 expression was identified by Western blot and qRT-PCR. The effect of inhibition of SOX2 on the content of breast cancer stern cells was detected by growth curve and flow cytometry. Results： The lentivirus-mediated SOX2 shRNA was ob- tained, MCF-7 cells and T47D cells stably expressing SOX2 shRNA were constructed, and the percentage of CD44＋/CD24-/low and ALDH＋ cells decreased significantly. Conclusion： After knockdown of SOX2, the content of breast cancer stem cells was inhibited.

关 键 词：SOX2 乳腺癌干细胞 CD44+/CD24-/low 乙醛脱氢酶 

分 类 号：Q78[生物学—分子生物学]