RNAi抑制STAT3对胶质瘤干细胞的生长抑制作用

The inhibition study of STAT3 siRNA on glioblastoma stem cells

目的构建STA33的RNAi载体干扰STA33的表达，在体内外研究对胶质瘤干细胞的影响。方法构建STA33基因shRNA的腺病毒表达载体，感染人胶质瘤干细胞。采用RT—PCR、Western blot检测STA33基因的干扰效果，用MTT和流式细胞仪检测干细胞的增殖、凋亡，并观察对荷瘤裸鼠的影响。结果转染重组腺病毒Ad—STAT3 siRNA后，胶质瘤干细胞的STAT3的mRNA表达率为（41．5±7．3）％，蛋白表达率为（31．2±6．4）％，细胞的增殖受到抑制，凋亡率为（23．8±6．1）％，同时荷瘤裸鼠的肿瘤生长受到抑制，生存期延长。结论重组腺病毒介导的STA33 RNAi可显著抑制靶基因的表达与活化，显著抑制胶质瘤干细胞的生长功能，为基于胶质瘤干细胞的功能治疗提供了基础理论。

Objective To construct STAT3 siRNA to interfere the expression STAT3, and to observe the effect on glioblastoma stem cells. Method The adenovirus shRNA expression vector targeting STAT3 gene was constructed and transduced into glioblastoma stem cells. The gene silencing efficiency was monitored by RT - PCR and Western blot analysis. The proliferation and apoptosis of glioblastoma stem cells were detected by MTT assays and flow cytometry. In addition, the effect of tumor bearing mice was also observed. Results After the adenovirus STAT3 siRNA transfection, the level of mRNA decreased to （41.5 ± 7.3 ） % , the level of protein decreased to （ 31.2 ±6. 4 ） % , the cell proliferation was inhibited, the cell apoptosis ratio increased to （ 23.8 ± 6. 1 ） %. Additionly, the adenovirus STAT3 siRNA could inhibit the tumor growth and improve the lifespan of mice. Conclusions The adenovirus shRNA targeting STAT3 gene could significantly inhibit the expression and activation of STAT3, and suppress the proliferation of glioblastoma stem cells ,which provide an ideal therapeutic strategy for treatment of glioblastoma stem cells.