隐丹参酮对人乳腺癌细胞MDA-MB-231转移的影响及其分子机制  

Anti-metastatic Effect of Cryptotanshinone on MDA-MB-231 Breast Cancer and Its Underlying Mechanism

作　　者：朱智杰[1,2] 刘兆国[1,2] 周梁[1,2] 田超[1,2] 陶丽[1,2] 韦忠红[1,2] 陆茵[1,2] 王爱云[1,2] 陈文星[1,2] 

机构地区：[1]南京中医药大学药学院,南京210046 [2]江苏省中药药效与安全性评价重点实验室,南京210046

出　　处：《中国实验方剂学杂志》2014年第11期160-165,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目资助(81173174);教育部博士点基金(20113237110008)

摘　　要：目的：探究隐丹参酮对人乳腺癌细胞（MDA-MB-231）体内、外转移的影响及其作用机制。方法：体外采用溴脱氧尿苷（Brdu）掺入免疫荧光实验考察隐丹参酮（0．5～8μmol·L^-1）作用24h对MDA-MB-231细胞（密度5×10^5／孔）增殖的影响；应用划痕和Transwell实验观察隐丹参酮（0．5～8μmol·L^-1）作用24h对肿瘤细胞（密度1×10’／孔）迁移的影响；采用Westernblot的方法考察隐丹参酮（0．5～8μmol·L^-1）作用24h对上皮间质转化相关蛋白E-钙黏素和N-钙黏素表达的影响；利用荧光素酶报告基因系统观察隐丹参酮（0．5～8μmol·L^-1）作用6h对MDA-MB-231细胞（密度为1×10^5／孔）核因子-κB（NF-κB）转录活性的影响；体内再应用斑马鱼肿瘤转移模型研究隐丹参酮（0．125～2μmol·L^-1）作用6天对人乳腺癌MDA-MB-231细胞转移数目及转移距离的作用来考察其对体内肿瘤转移的影响。结果：在体外，隐丹参酮在0．5～8μmol·L^-1可以剂量依赖性地抑制MDA-MB-231BrdU阳性细胞的比例，抑制划痕实验和Transwell实验中迁移细胞的数目，增加E-钙黏素的表达，同时降低N-钙黏素的表达，并抑制NF-KB的相对启动子活性。在体内，隐丹参酮在0．125～2μmol·L^-1可以剂量依赖性地减少MDA-MB-231乳腺癌细胞远端转移的数目，降低转移的最远距离。结论：隐丹参酮在体内外均有显著的抗乳腺癌转移作用，在体外对肿瘤细胞增殖还有一定作用，其机制可能是通过抑制NF-κB转录活性从而逆转MDA-MB-231细胞的上皮间质转化起作用的。Objective： To investigate the anti-metastatic effect of cryptotanshinone on human breast cancer and its possible mechanisms. Method： The ability of cryptotanshinone on proliferation was detected after labeling with bromodeoxyuridine （BrdU） by immunofluorescence staining. We furher used wound healing assay and Transwell migration assay to investigate the effect of cryptotanshinone on cell mobility in vitro. Western blot was used to measure the protein expression related to epithelial mesenchymal transition （EMT）. Effect ofcryptotanshinone on nuclear factor （NF） -κB transcriptional activity was detected by Dual-Luciferase gene reporter assay system. Furthermore, we used transgenic zebrafish tumor model to illustrate the anti-metastatic effect of cryptotanshinone in vivo. Result： Cryptotanshinone showed a great inhibitory ability on cell proliferation and migration in vitro. Cryptotanshinone also does-dependently increased the expression of E-cadherin but decreased the expression of N-cadherin. The transcriptional activity of NF-KB was prohibited by cryptotanshinone as well. Moreover, cryptotanshinone inhibited MDA-MB-231 tumor metastasis in a dose dependent manner in zebrafish embyos by decreasing the number of disseminated foci from tumor mass as well as maximal distance of metastasis. Conclusion： Cryptotanshinone showed both anti-metastatic effect in vivo and in vitro. The proliferation of MDA- MB-231 was also inhibited by cryptotanshinone. The underlying anti-metastatic mechanism of cryptotanshinone on MDA-MB-231 breast cancer cells involved reversion of EMT via downregulating transcriptional activity of NF-κB.

关 键 词：隐丹参酮 乳腺癌 肿瘤转移 上皮间质转化 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]