目的 研究白頭翁皂苷D對結腸癌上皮間質轉化（epithelial-mesenchymal transition，EMT）的影響，并基于無翅型MMTV整合位點家族（wingless type MMTV integration site family，Wnt）/β-連環(huán)蛋白（β-catenin）信號通路研究其作用機制。方法 通過MTT細胞增殖實驗研究白頭翁皂苷D對人結腸癌LoVo細胞的影響，細胞劃痕實驗研究白頭翁皂苷D對LoVo細胞遷移的影響，Transwell細胞侵襲實驗研究白頭翁皂苷D對LoVo細胞侵襲的影響，錨定非依賴性克隆形成實驗研究白頭翁皂苷D對LoVo細胞惡性轉化的影響；采用實時熒光定量聚合酶鏈式反應（real-time fluorescent quantitative polymerase chain reaction，qRT-PCR）檢測LoVo細胞EMT相關基因和Wnt/β-catenin信號通路基因的mRNA水平，采用Western blotting法檢測LoVo細胞EMT相關蛋白和Wnt/β-catenin信號通路蛋白的表達水平。結果 白頭翁皂苷D顯著抑制LoVo細胞增殖活性、遷移、侵襲和克隆形成，與對照組比較，白頭翁皂苷D可顯著降低EMT相關的波形蛋白（Vimentin）和蝸牛蛋白（Snail）的mRNA和蛋白表達（P＜0.05、0.01）、升高E-鈣黏蛋白（E-cadherin）的mRNA和蛋白的表達（P＜0.01），降低Wnt/β-catenin信號通路中Wnt3a、β-catenin、T細胞因子4（T-cell factor 4，TCF4）和原癌基因（myelocytomatosis viral oncogene homolog，c-Myc）的mRNA和蛋白表達（P＜0.05、0.01），升高糖原合成激酶-3β（glycogen synthase kinase-3β，GSK-3β）的mRNA和蛋白表達（P＜0.05、0.01）。白頭翁皂苷D抑制LoVo細胞遷移和調節(jié)E-cadherin與Vimentin表達的作用可被Wnt/β-catenin信號通路抑制劑IWR-1協(xié)同增強，也被Wnt/β-catenin信號通路激動劑SKL2001拮抗減弱。結論 白頭翁皂苷D可抑制結腸癌的EMT，其機制在于抑制Wnt/β-catenin信號通路活性。

Objective To investigate the effects of Pulsatilla saponin D (PSD) on the epithelial-mesenchymal transition (EMT) of colorectal cancer, and explore its action mechanisms based on the wingless type MMTV integration site family (Wnt)/β-catenin signaling pathway. Methods Effect of PSD on the cell viability of LoVo cells was determined with the MTT assay. Effect of PSD on the migration of LoVo cells was assessed with the scratch wound healing assay. Effect of PSD on the invasion of LoVo cells was examined with the Transwell invasion assay. Effect of PSD on the malignant transformation of LoVo cells was appraised with the anchorage-independent colony formation assay. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) analysis was carried out to detect the effect of PSD on the mRNA levels of EMT-related genes and the genes of Wnt/β-catenin signaling pathway in LoVo cells. Western blotting was performed to measure the effect of PSD on the expression levels of EMT-related proteins and the proteins of Wnt/β-catenin signaling pathway in LoVo cells.Results PSD significantly inhibited the proliferation activity, migration, invasion and colony formation of LoVo cells. Compared with the control group, PSD could reduce the mRNA and protein levels of EMT-related Vimentin and Snail (P < 0.05, 0.01), increased the mRNA and protein levels of E-cadherin (P < 0.01). It decreased the mRNA and protein levels of Wnt3a, β-catenin, T-cell factor 4 (TCF4), and myelocytomatosis viral oncogene homolog (c-Myc) in Wnt/β-catenin signaling pathway (P < 0.05, 0.01), increased the mRNA and protein levels of glycogen synthase kinase-3β (GSK-3β) (P < 0.05, 0.01). Furthermore, the inhibition of cell migration and the regulation of the expressions of E-cadherin and vimentin in LoVo cells by PSD were synergized by Wnt/β-catenin signaling pathway inhibitor IWR-1, and antagonized by Wnt/β-catenin signaling pathway activator SKL2001. Conclusion PSD may suppress the EMT of colorectal cancer, potentially by inhibiting the activity of Wnt/β-catenin signaling pathway.

R285.5

國家自然科學基金資助項目（81573813）;四川省自然科學基金面上項目（2023NSFSC0653）