Resveratrol抑制人類乳腺癌細胞株MDA-MB-231的生長、移行和侵襲能力

Abstract

中文摘要 Resveratrol (白藜蘆醇, 3,4,5'-trihydroxy-trans-stilbene)是一種存在植物內天然的多酚類 (polyphenol)化合物，本實驗採用乳腺癌細胞株 (MDA-MB-231cell line)進行細胞培養的研究，探討resveratrol對MDA-MB-231 cells生長、細胞移行與侵襲的影響，實驗結果如下(1) MTT assay實驗結果顯示: 細胞經resveratrol處理24小時後，高濃度的resveratrol (30, 50, 70μΜ)與未給予藥物之控制組相較，會促使MDA-MB-231細胞存活率降低 (83％, 74％, 22％)，達到顯著差異，且細胞株處理不同時間 (24, 48, 72小時)，細胞生長比例皆低於控制組，均達到顯著差異。西方墨點分析顯示: resveratrol可透過減少抗細胞凋亡bcl2蛋白表現及增加促細胞凋亡caspase3蛋白表現，而造成細胞凋亡；(2) 細胞移行分析與細胞傷口癒合分析結果顯示: 不同濃度的resveratrol (30, 50μΜ)處理下，細胞的移行能力降低，統計上達到顯著的差異。免疫細胞螢光染色結果顯示: resveratrol (30, 50μΜ)可以正向調控 MDA-MB 231細胞株E-cadherin的表現，以及抑制細胞內皮生長因子(VEGF)的表現。(3) 細胞侵襲分析結果顯示: 不同濃度的resveratrol (30、50μΜ)處理下，細胞侵襲的細胞數下降，統計上達到顯著的差異。西方墨點分析結果顯示: 不同濃度的resveratrol ( 30, 50μΜ)處理下，使基質金屬蛋白酶9 (MMP9)的表現下降。另外，利用明膠蛋白酵素電泳法證實經resveratrol ( 30, 50μΜ)處理下，細胞分泌至細胞外的MMP9之活性降低。 綜合以上結果顯示: resveratrol抑制乳腺癌MDA-MB-231 cells生長、移行與侵襲的過程可能透過抑制bcl2、VEGF與MMP9蛋白的表現及活化caspase3與E-cadherin的表現，進而達到抑制MDA-MB-231 cells生長、移行與侵襲的目的。

英文摘要 Resveratrol is a natural polyphenol compound within the plant. We used MDA-MB-231 breast adenocarcinoma cell line as a model to investigate the effects of resveratrol on proliferation, migration, and invasion of breast cancer cell. By MTT assay, we found that after treatment with 30, 50, 70μΜ resveratrol for 24 hours, the proliferation ratio of MDA-MB-231 cells decreased to 83％, 74％, 22％. The results of time course study also (24, 48, 72hours) revealed that the proliferation ratio of 30, 50, 70μΜ resveratrol-treated MDA-MB-231 cells were decreased. Western blotting showed that resveratrol caused cell apoptosis resulting from the suppression of bcl2 protein expression and activation of caspase3 protein expression. In cell migration assay and wound healing assay revealed that MDA-MB-231 cells treated with resveratrol decrease the migration of cells. The effects of resveratrol on the expression of E-cadherin and VEGF by MDA-MB-231 were analyzed by fluorescence immunocytochemistry stain. Results indicate that resveratrol up-regulated E-cadherin expression and down-regulated VEGF expression. By cell invasion assay, we found that after being treated with resveratrol, the cells decreased in invasion ability. Western blotting showed that resveratrol caused the suppression of MMP9 expression. Besides, the activation of MMP9 secreted to the extracellular fluid, analyzed by gelatin zymography, was decreased in MDA-MB-231 cell line after being treated with resveratrol. The results obtained from the present study showed that resveratrol might play an important role in the suppression of the proliferation, migration and invasion of breast cancer cell line of MDA-MB-231 cells through the inhibition of bcl2, VEGF and MMP9 and activation of caspase3 and E-cadherin.