敲低Yes相关蛋白表达对人脑胶质瘤细胞SNB19生长的抑制作用

Inhibitory effect of knocking-down Yes-associated protein for the growth of SNB19 glioma cells

目的探究敲低Yes相关蛋白（YAP）对人脑胶质瘤细胞SNBl9细胞生物学特征的影响。方法采用YAP干扰RNA（YAPsiRNA）敲低SNBl9细胞中YAP的表达，Westernblot法鉴定肿瘤细胞中YAP是否已被敲低；噻唑蓝（MTY）比色法测定YAP敲低后胶质瘤细胞的增殖能力；Transwell实验检测胶质瘤细胞侵袭能力的变化；流式细胞术和AnnexinV标记法分别检测胶质瘤细胞周期及凋亡的变化。应用统计软件SPSS18．0进行统计学处理。结果Westernblot证实转染YAPsiRNA可有效敲低SNB19细胞中Yes相关蛋白的表达；敲低Yes相关蛋白的表达可以抑制胶质瘤细胞增殖活性，细胞存活率由（100．00±0．00）％下降为（52．32±3．10）％（F=33．00，P〈0．01），细胞周期阻滞于G0-G1期（F=8．76，P〈0．01），细胞侵袭能力明显受抑，穿膜细胞数由（163．20±10．10）个下降至（37．71±2．52）个（F=282．05，P〈0．01），凋亡增加，凋亡率由（3．56±0．35）％增加至（18．99±0．66）％（F=931．99，P〈0．01）。结论敲低胶质瘤细胞SNB19中YAP表达可抑制肿瘤细胞增殖和侵袭，诱导细胞凋亡；YAP可成为人脑胶质瘤基因治疗的潜在靶点。

Objective To investigate the effect of knocking-down Yes-associated protein （YAP） on the biologi- cal characteristics of SNB19 glioblastoma cell. Methods The expression of YAB in SNB19 was knockdown by YAB small interfering RNA （YABsiRNA）. The downregulafion of YAP expression was identified by Western blot analysis. The proliferative ability of cell＇was determined by methyl thiazoyl terazolium （MTF）. The invasive ability of cell was examined by Transwell assay. Flow cytometry and Annexin V staining were used to detect the cell cycle and apoptosis respectively. The results were analyzed by the statistical software SPSS18.0. Results ~ expression of YAP in the cells transfected with YAPsiRNA was significantly reduced. The cell proliferation activity of SNB19 cells was inhibited, which decreased from （ 100. 00 ±0.00） % to （52.32± 3.10） % （ F = 33. （30, P 〈 0. 01 ）. Tile cell cycle was arrested in G0-Gl phase （F =8.76, P 〈0.01 ）. The cell invasive ability was attenuated apparently, which decreased from （ 163.20±10. 10） to （37.71 ±2.52） （F =282.05, P 〈0.01）. The apoptosis ratio of the tumor cell which transfected with YAPsi- RNA was increased from （3.56 ± 0.35） % to （ 18.99 ± 0. 66） %, （ F = 931.99, P 〈 0. 01 ）. Conclusion Knocking-down YAP expression in glioma cells could inhibit the proliferative activity and invasive ability of SNB19 cell and could induce cell apoptosis. YAP could be served as a potential target for the gene therapy of glioma.