摘要
目的研究雷帕霉素(rapamycin,RAPA)在体外对上皮鳞癌细胞系A431的增殖、迁移以及侵袭能力的影响。方法体外培养人上皮鳞癌细胞A431,通过噻唑蓝(MTT)实验检测不同浓度的RAPA(5、10、20 nmol·L-1)对A431增殖的影响,并筛选出最适浓度。依此浓度分别通过划痕、Transwell试验检测A431的迁移和侵袭能力变化。经RAPA处理后,采用逆转录-聚合酶链反应(RT-PCR)和Western blot检测A431中骨桥蛋白(osteopontin,OPN)的mRNA和蛋白的表达水平的变化。结果 MTT实验结果显示不同浓度RAPA对A431增殖有抑制作用,且表现浓度依赖性。RAPA作用于A431细胞48 h后,划痕实验及Transwell实验显示A431的迁移和侵袭能力受到明显抑制。RT-PCR和Western blot结果都显示RAPA处理的A431细胞中OPN的表达下调。结论 RAPA可降低鳞癌细胞A431的增殖、迁移以及侵袭能力,其机制可能与RAPA抑制OPN的表达有关。
Aim To investigate the effect of rapamycin on proliferation, migration, and invasion of squamous cell carcinoma A431 cells in vitro. Methods Human squamous cell carcinoma A431 cells were cultured invitro. MTr assay was used to investigate proliferation of A431 cells treated by rapamycin at different concentra- tions of （5,10,20 nmol· L-1）. Wound healing and transwell assay were performed to detect migration andinvasion of A431 cells treated by rapamycin, respec- tively. Reverse transcription PCR （RT-PCR） and Western blot were used to determine the expression of the osteopontin （OPN） in A431 cells at mRNA level and protein level, respectively. Results Rapamycin significantly increased the inhibitory rates of prolifera- tion and inhibited the migration and invasion of A431 cells in vitro. Furthermore, rapamycin treatment re-duced sions the expression of OPN in A431 cells. Conclu- Rapamycin can inhibit the migration and inva- sion of A431. The intrinsic mechanism of rapamycin might be related to the down-regulation of the OPN ex- pression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2014年第4期550-554,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81172591)
