紫草素下调Runx2转录活性对乳腺癌细胞体外运动能力的影响  

Influence of shikonin for the motility in vitro of breast cancer cells via down-regulating transcriptional activity of Runx2

作　　者：王芳[1] 吕红[2] 贾娟娣 韩伟[2] 付剑江[3] 

机构地区：[1]解放军总医院肾脏科,北京100853 [2]江西中医药大学网络与教育技术中心,江西南昌330004 [3]江西中医药大学药学院,江西南昌330004

出　　处：《中国医药导报》2016年第18期4-8,共5页China Medical Herald

基　　金：国家自然科学基金资助项目(81160530;81260656;81560639)

摘　　要：目的探讨紫草素(SKN)对乳腺癌细胞体外运动能力的影响。方法利用Oris TM细胞迁移试剂盒和重组基底膜侵袭实验检测SKN对乳腺癌细胞MDA-MB-231细胞体外迁移能力和侵袭能力的影响。采用明胶酶谱法观察SKN对MDA-MB-231细胞分泌基质金属蛋白酶(MMPs)的影响。通过染色质免疫共沉淀(Ch IP)和Western blot实验检测SKN对MDA-MB-231细胞内Runx2转录活性及表达的影响。结果给药后,20μmol/L SKN可显著抑制表皮生长因子诱导的细胞迁移作用;重组基底膜侵袭实验中,SKN 10μmol/L和20μmol/L剂量组的侵袭细胞数分别为(514.3±76.8)、(426.3±87.0)个,与空白组[(738.5±126.1)个]比较差异有统计学意义(P<0.05);明胶酶谱实验显示,SKN可下调基质金属蛋白酶-9(MMP-9)的活性;进一步Western blot实验结果表明,MDA-MB-231细胞的MMP-9表达也显著降低(P<0.01)。Ch IP实验结果显示,SKN可明显下调Runx2与MMP-9基因启动子区内特定序列的结合能力,而Western blot结果发现,SKN可显著抑制磷酸化Runx2的表达(P<0.01)。结论 SKN对MDA-MB-231细胞体外运动功能具有显著抑制作用,这种作用与其抑制Runx2活化、下调Runx2转录活性、抑制MMP-9表达有关。Objective To investigate the influence of shikonin（SKN） for the motility in vitro of breast cancer cells.Methods The influence of SKN for the migration and invasion in vitro of MDA-MB-231 cells was detected by OrisTM migration assay and reconstituted basement membrane invasion assay. Gelatin zymography was used to observe the effects of SKN on MDA-MB-231 cells in secreting matrix metalloproteinase（MMPs）. Chromatin immunoprecipitation（Ch IP） assay and Western blot test were used to detect the influence of SKN for transcriptional activity and expression of Runx2 in MDA-MB-231 cells. Results After administration, 20 μmol/L SKN could significantly inhibit the cell migration induced by epidermal growth factor; in the reconstituted basement membrane invasion assay, the number of invaded cells of 10, 20 μmol/L of SKN was（514.3±76.8）,（426.3±87.0） respectively, which had statistically significant differences compared with that of blank group（738.5±126.1）（P〈 0.05）; Gelatin zymography showed that SKN could down-regulate the activity of matrix metalloproteinase-9（MMP-9）; further Western blot showed that, the expression of MMP-9 was decreased significantly（P〈 0.01）. The Ch IP assay showed that SKN could down-regulate the combining capacity of Runx2 with the specific sequences of HTT gene-linked polymorphic region in MMP-9, while the results of Western blot found that SKN could inhibit the expression of phosphorylation Runx2（P〈 0.01）. Conclusion SKN has significant inhibitory effect for the motility in vitro of MDA-MB-231 cells, which may be related to inhibiting excitation of Runx2, down-regulating the transcriptional activity of Runx2, inhibiting the expression of MMP-9.

关 键 词：紫草素 体外运动能力 基质金属蛋白酶-9 RUNX2 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]