摘要
目的 探讨ErbB3在骨肉瘤细胞Saos-2中的表达及其意义。方法 实时定量PCR检测骨肉瘤细胞Saos-2和正常人类成骨细胞N704中ErbB3 mRNA的表达水平，Western blotting检测ErbB3蛋白表达水平。使用短发夹RNA（shRNA）构建ErbB3表达沉默的Saos-2细胞，检测0-3 d内Saos-2细胞数目。测定ErbB3表达沉默和正常Saos-2细胞在0-100 μmol/L紫杉醇下48 h存活率。结果 相对于正常人类成骨细胞N704，骨肉瘤细胞Saos-2的ErbB3 mRNA表达量增加（4.15±0.04）倍，差异有统计学意义（t=7.31，P＜0.05），且ErbB3蛋白水平显著增加。ErbB3沉默能降低Saos-2细胞的增殖水平，培养第3天ErbB3沉默的Saos-2细胞数目为（2.22±0.29）万个，而未沉默细胞数目为（4.58±0.41）万个，差异有统计学意义（t=8.23，P＜0.05）。此外，ErbB3沉默能降低Saos-2细胞对紫杉醇的耐受，给药紫杉醇20 μmol/L时，ErbB3沉默组细胞存活率为（43.2±4.7）%，而未沉默组存活率为（61.4±5.9）%，差异有统计学意义（t=6.74，P＜0.05）。结论 骨肉瘤细胞Saos-2高度表达ErbB3，ErbB3的表达可增强Saos-2细胞的增殖能力，同时增强Saos-2细胞对紫杉醇的耐受。
Objective To explore the expression of ErbB3 in osteosarcoma cell lines Saos-2 and its significance. Methods Real-time quantitative PCR was used to detected ErbB3 mRNA expression in osteosarcoma cell lines Saos-2 and normal human osteoblasts cell lines N704, and Western blotting was used to detected ErbB3 protein expression. Short hairpin RNA was used to construct ErbB3 knockdown cells and the cell count in 0-3 d was detected. The 48 h survival rates of ErbB3 knockdown cells and normal Saos-2 cells were detected in the presence of 0-100 μmol/L paclitaxel. Results Real-time quantitative PCR showed significant enhanced ErbB3 mRNA expression in Saos-2 cells compared with N704 [（4.15±0.04） times, t=7.31, P〈0.05], and Western blotting showed significant enhanced ErbB3 expression in Saos-2 cells compared with N704. As compared with normal Saos-2 cells, ErbB3 knockdown could reduce the Saos-2 cells proliferation, at the first three days in culture, the Saos-2 cell number of ErbB3 silent was （22.2±2.9） thousand, and the control was （45.8±4.1） thousand, with statistical significance （t=8.23, P〈0.05）. Moreover, ErbB3 knockdown could reduce the Saos2 cells tolerance to paclitaxel, when treated with 20 μmol/L paclitaxel, surviving cells in ErbB3 silent group was （43.2±4.7）%, and the control was （61.4±5.9）%, with statistical significance （t=6.74,P〈0.05）. Conclusion ErbB3 highly expresses in Saos-2 cells, ErbB3 expression can enhance the proliferation of Saos-2 cells and the tolerance of Saos-2 cells to paclitaxel.
出处
《国际肿瘤学杂志》
CAS
2016年第8期593-596,共4页
Journal of International Oncology
关键词
骨肉瘤
RNA干扰
紫杉醇
Osteosarcoma
RNA interference
Paclitaxel
