芹菜素藉由影響Pin 1和Wnt/β-catenin信號路徑抑制三陰性乳癌的增生和侵襲

Apigenin suppresses cell proliferation and invasion via inhibiting Pin 1 and Wnt/β-catenin signaling pathways in triple-negative breast cancer

廖泯嘉(Min-Chia Liao)

指導教授 ： 陳威仁

中山醫學大學/醫學科技學院/生物醫學科學學系/碩士(2021年)

https://doi.org/10.6834/csmu202100158

摘要

背景：WNT /β-catenin信號通路的異常在人類腫瘤發生中起重要作用。 β-catenin的功能異常會導致下游基因（如細胞週期蛋白D1）的過度表達，並加速人類癌症的細胞週期進程。芹菜素是一種富含於蔬菜和水果的類黃酮。據報導，芹菜素具有抗癌活性，具有低的細胞毒性和誘變活性，但芹菜素抑制三陰性乳腺癌細胞增殖和侵襲的機制仍不清楚。為此，我們進一步探討芹菜素是否通過調節Wnt /β-catenin信號通路和Pin 1蛋白來抑制三陰性乳腺癌的增殖和epithelial-to-mesenchymal transition (EMT)。
方法：在這項研究中，我們進行了MTT，傷口癒合和transwell入侵試驗，以分別評估細胞毒性，增殖，細胞遷移和侵襲。接著進行西方墨點法監測參與WNT /β-catenin途徑的活性分子的蛋白質水平。
結果：首先我們看到在經過芹菜素處理後，細胞的型態從移動性強的梭狀型態變成低移動性的上皮細胞型態，代表芹菜素能降低細胞的移動能力。另外，藉由MTT assays我們發現芹菜素具有輕微抑制細胞增生的效果。除此之外，由傷口癒合和transwell入侵試驗的結果表明，芹菜素以濃度依賴性方式抑制細胞遷移和侵襲。 而Western蛋白分析則顯示芹菜素降低了Pin 1和β-catenin蛋白的水平，並同時抑制了EMT相關蛋白的表達。
結論：芹菜素可以通過抑制Pin 1蛋白來調節β-catenin在核內的表現量，進而抑制EMT。因此，我們推測芹菜素可以作為三陰性乳腺癌患者的輔佐治療藥劑。

關鍵字

芹菜素

並列摘要

Background: Abnormal WNT/ β-catenin signaling pathway plays an important role in human tumorigenesis. Aberrant β-catenin function leads to overexpression of its effector gene such as cyclin D1 and accelerates cell cycle progression in human cancers. Apigenin, a kind of flavonoids abundant in vegetables and fruits, has been reported to possess anti-cancer activity with low cytotoxicity and mutagenic activity, but the mechanism by which apigenin inhibits triple-negative breast cancer cell proliferation and invasion still unclear. To this end, we investigated whether apigenin inhibited proliferation and epithelial-to-mesenchymal transition (EMT) of triple-negative breast cancer via inhibition of Pin 1 and the Wnt/β-catenin signaling pathways.
Methods: In this study, we conducted MTT, wound healing and, transwell invasion assays, to evaluate cytotoxicity, proliferation, cell migration, and invasion, respectively. Western blotting was performed to monitor the protein levels of active molecules involved in WNT/ β-catenin pathway.
Results: First, we obseverd that the cell type changed from a highly mobile spindle-shaped type to a low-mobility epithelial cell type after apigenin treatmen, which means that apigenin can reduce the mobility of cells. In addition, we found that apigenin has a slight inhibitory effect on cell proliferation by MTT assays. Then the results of wound healing and transwell invasion assays demonstrated that apigenin inhibited cell migration and invasion in a dose-dependent manner. Western blot analysis revealed that apigenin decreased the Pin 1 and β-catenin protein levels, and concomitantly suppressed the expression of EMT-associated proteins.
Conclusion: Apigenin can regulate β-catenin level to inhibit EMT through inhibiting Pin 1. We thus suggest that apigenin may act as a potential adjunctive therapeutic agent for triple-negative breast cancer patients.

並列關鍵字

Apigenin

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