目的 揭示白及Bletilla striata提取物抗肿瘤的有效成分和潜在分子机制。方法 首先通过生物活性追踪方法鉴定白及中抗肿瘤活性成分。采用CCK-8法检测化合物对膀胱癌5637细胞的毒性，进一步筛选出最优化合物。激光共聚焦显微镜观察最优化合物对膀胱癌细胞凋亡诱导。Transwell侵袭实验检验最优化合物对膀胱癌细胞侵袭的影响，Western blotting检测细胞内上皮钙黏蛋白（epithelial cadherin，E-cadherin）、神经钙黏蛋白（neural-cadherin，N-cadherin）、波形蛋白（‌Vimentin）、桥粒斑蛋白（desmoplakin，DSP）、胞质紧密粘连蛋白1（zona occludens 1，ZO-1）、细胞外调节蛋白激酶（extracellular regulated protein kinases，ERK）、磷酸化细胞外蛋白调节激酶（phospho-extracellular signal-regulated kinase，p-ERK）蛋白表达水平。结果 从白及干燥块茎中分离鉴定出12个菲类化合物，分别为2,7-二羟基-4-甲氧基-9,10-二氢菲（1）、红门兰醇（2）、次苦参素（3）、2,7-二羟基-1-(对羟基苄基)-4-甲氧基-9,10-二氢菲（4）、4,7-二羟基-2-甲氧基-9,10-二氢菲（5）、2,7-二羟基-4-甲氧基菲（6）、2-羟基-4,7-二甲氧基菲（7）、8-(4-(甲酰氧基)苄基)-7-甲氧基-9,10-二氢菲-2,5-二基二甲酸酯（8）、白及联菲A（9）、7-羟基-2-甲氧基-9,10-二氢菲-1,4-二酮（10）、7-羟基-2-甲氧基-9,10-二氢菲-1,4-二酮（11）、7-羟基-2-甲氧基-菲-1,4-二酮（12）。化合物1在40 μg/mL质量浓度下，对膀胱癌细胞5637抑制率为（53.25±1.35）%，在10、40、80 μg/mL质量浓度下，侵袭细胞数量显著减少，分别为（21.31±5.61）%、（54.72±0.98）%和（83.12±0.98）%。结论 化合物8、10～12为首次从该植物中分离得到。化合物1可抑制ERK信号通路的激活来阻止膀胱癌5637细胞的增殖，诱导细胞的凋亡，并通过EMT进程抑制人膀胱癌5637细胞的侵袭。

Objective To reveal the anti-tumor active components and molecular mechanisms of Bletilla striata. Methods The toxicity of the compound to bladder cancer cell line 5637 was detected by CCK-8 method. The apoptosis of bladder cancer cells induced by coelonin solution was observed. Transwell invasion assay was used to examine the effect of coelonin on the invasion of bladder cancer cells, and western blot assay was used to detect the expression levels of E-cadherin, N-cadherin, vimentin, desmoplakin, Zona occludens 1, extracellular regulated protein kinases (ERK), phospho-extracellular signal-regulated kinase (P-ERK) protein in cells. Results A total of 12 compounds were isolated from B. striata rhizome, respectively coelonin (1), orchinol (2), lusianthridin (3), shancidin (4), 4,7-dihydroxy-2-methoxy-9,10-dihydrophenanthrene (5), flavanthrinin (6), 2-hydroxy-4,7-dimehoxy-phenanthrene (7), 8-(4-(formyloxy)benzyl)-7-methoxy-9,10-dihydrophenanthrene-2,5-diyl diformate (8), blestriarene A (9), ephemeranthoquinone (10), ochrone A (11), densiflorol B (12). Compound 1 at 40 μg/mL showed a (53.25 ±1.35)% inhibition rate on bladder cancer cell line 5637. At different concentrations (low, medium and high), the number of invasive cells decreased significantly, which were (21.31 ±5.61)%, (54.72 ±0.98)% and (83.12 ±0.98)%, respectively. Conclusion Compounds 8, 10—12 are identified from B. striata genus for the first time. Coelonin can inhibit the activation of ERK signaling pathway to prevent the proliferation of bladder cancer 5637 cells, and inhibit the invasion of cells through EMT process. The antitumor activity of coelonin, as the main active ingredient of B. striata, is worthy of further study.

R284.1

湖南省教育厅优秀青年项目（23B0751）;湖南省区域联合基金项目（2024JJ7198）