摘要
目的 研究姜黄素对侵袭信号传导和转录激活因子3（signal transducer and activator of transcription 3,STAT3）表达的调控,初步探讨姜黄素在防治皮肤鳞状细胞癌侵袭转移中可能存在的新的作用机制.方法 以高侵袭性皮肤鳞癌细胞株A431为研究对象,采用不同剂量姜黄素处理.MTT法检测5,10,15,20,25,30,35,40和50 μmol/L姜黄素作用24,48和72 h对细胞的毒性作用;Transwell小室侵袭试验法检测5,10和15μmol/L姜黄素对细胞的侵袭能力,黏附试验法检测对细胞的黏附能力;Western blot法检测5,10和15 μmol/L姜黄素作用后STAT3蛋白表达水平的影响,RT-PCR法检测对细胞中STAT3基因mRNA转录水平的影响.结果 姜黄素浓度＜15 μmol/L,作用时间24 h时,对细胞的生长抑制呈时间和剂量依懒性（P＞0.001）,当姜黄素浓度15 μmol/L,作用时间为24 h时,对细胞无明显毒性作用,细胞存活率85％;细胞侵袭能力和黏附能力随着姜黄素浓度的增加而逐渐降低,以15 μmol/L姜黄素浓度处理效果最明显（P＞0.05）;姜黄素可明显抑制STAT3基因mRNA转录水平,其抑制作用与剂量呈高度依懒性（P＞0.05）.结论 姜黄素可通过抑制STAT3信号通路的活化及该信号通路靶基因STAT3的表达降低皮肤鳞癌A431细胞的侵袭性.
Objective To investigate the regulatory effect of curcumin on expression of signal transducer and activator of tran- scription 3 （STAT3） in Skin squamous cell carcinoma tissues as well as the possible mechanism of curcumin in prevention and treatment of skin squamous cell carcinoma. Methods Highly invasive A431 cells were treated with curcumin at various dosages. The cytotoxic effects of treatment with 5,10,15,20,25,30,35,40 and 50 μmol/L curcumin for 24,48 and 72 h on A431 cells were measured by MTT assay. The invasion abilities of cells treated with 5,10 and 15 μmol/L curcumin were measured by Transwell test,while the adhesive abilities by cell adhesion assay. The effect of 5,10 and 15 μmol/L curcumas on expression levels of STAT3 were determined by western blot, while those on transcription level of STAT3 mRNA by RT- PCR. Results The treatment with eurcumin at a dosage of more than 15 μmol/L for more than 24 h inhibited the growth of A431 cells in time-and dose-dependent patterns （P（0. 001）. The treatment with curcumin at a dosage of not less than 15μmol/L for 24 h showed no significant cytotoxic effect on the cells,while the survival rate of ceils was more than 85 %. The invasion and adhesive abilities of cells decreased gradually with the increasing curcumin concentration. The curcumin at a concentration of 15 μmol/L showed the strongest inhibitory effect on the two abilities （P〈0.05）. Curcumin showed signifi- cantly dose-dependent inhibitory effect on the transcription level of STAT3 mRNA （P〈0.05）. Conclusion Curcumin inhib- ited the invasive abilitiy of A431 cells by inhibiting the activation of STAT3 signal pathway and expression of STAT3 as a target gene of the pathway.
出处
《现代检验医学杂志》
CAS
2015年第5期62-65,69,共5页
Journal of Modern Laboratory Medicine
