目的 研究筋骨草总环烯醚萜对高转移三阴性乳腺癌细胞侵袭转移的作用及分子机制。方法 采用肿瘤细胞黏附血管内皮细胞实验、Transwell迁移和侵袭实验以及尾静脉注射诱导的实验性乳腺癌肺转移模型, 检测细胞黏附、运动、侵袭和转移能力。Western blot 法检测上皮-间质转化(epithelial-mesenchymal transitions, EMT)相关蛋白和胞外信号调节激酶(ERK)1/2丝裂原活化蛋白激酶(MAPK)通路蛋白的表达。结果 筋骨草总环烯醚萜50~200 mgL-1对MDA-MB-231细胞的黏附、迁移和侵袭能力有明显的抑制作用。连续给予200 mgkg -1筋骨草总环烯醚萜3周能明显抑制荷瘤动物肺转移。进一步研究发现, 筋骨草总环烯醚萜能显著降低p-ERK1/2蛋白表达;逆转乳腺癌细胞EMT, 主要表现在上皮性标志物E-cadherin和ZO-1表达增加, 间质性标志物vimentin表达降低。结论 筋骨草总环烯醚萜能有效地抑制高转移性乳腺癌细胞的侵袭和转移, 作用机制可能与其调控ERK1/2 MAPK通路, 逆转肿瘤细胞EMT有关。
Abstract
OBJECTIVE To investigate the effect of iridoids in Ajuga decumbens(ADI) on the metastasis of invasive breast cancer cells and related mechanisms. METHODS Several assays including cell-HUVEC adhesion assay were conducted, Transwell migration and Transwell invasion assay were conducted. In vivo antimetastatic effect of ADI was determined using experimental lung metastatic models induced by tail vain injection of 4T1-luc cells. Western blot analysis was employed to detect the expression of associated proteins of EMT and ERK1/2 MAPK signaling pathway. RESULTS The 50-200 mgL-1 ADI could significantly inhibit the cell-HUVEC adhesion, migration and invasion of MDA-MB-231 cells. Administration with 200 mgkg -1 dose of ADI markedly inhibited the lung tumor nodules. Treatment with ADI resulted in markedly inhibition of phosphorylation of ERK1/2 and the reversal EMT of breast cancer cells, which increased the levels of epithelial biomarkers, such as E-cadherin and ZO-1, and reduced the levels of mesenchymal biomarkers, such as vimentin. CONCLUSION These results indicates that ADI can inhibit breast cancer cell invasion by suppressing the ERK1/2 MAPK pathway as well as reversing the epithelial-mesenchymal transition.
关键词
乳腺癌 /
筋骨草总环烯醚萜 /
肿瘤转移 /
丝裂原活化蛋白激酶通路 /
上皮间质转化
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Key words
breast cancer /
iridoids in Ajuga decumbens /
MAPK pathway /
metastasis /
epithelial-mesenchymal transition
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81302981);中国中医科学院中药研究所基本科研业务费自主选题资助项目(ZZ2014004, ZXKT15021)
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