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目的观察小檗碱(Berberine,BBR)对鼻咽癌细胞埃兹蛋白(Ezrin)和磷酸化埃兹蛋白(phos-Ezrin)表达的影响以及其对细胞伪足形成和迁移的抑制作用,探讨BBR抗鼻咽癌转移可能的分子机制。方法以体外培养的鼻咽癌细胞株(CNE1)为研究对象,用MTT法分析BBR对CNE1细胞生长的影响,选用BBR细胞无毒性浓度(non-cytotoxic concentration,NCC)进行实验。免疫印迹(Western-blotting)分析CNE1Ezrin和phos-Ezrin的表达。用光镜CNE1观察细胞迁移能力,电子显微镜观察细胞伪足形成。Ezrin小干扰RNA(Ezrin-siRNA)转染阻断Ezrin蛋白表达,进一步确证BBR通过Ezrin抑制细胞伪足形成。结果MTT结果显示,BBR在5μM以上明显抑制CNE1增殖(P<0.05),浓度(0~5)μM对CNE1细胞生长增殖抑制作用不明显,为BBR对CNE1的细胞无毒性浓度(NCC)。BBR在NCC能明显抑制CNE1细胞phos-Ezrin表达,且呈浓度依赖性。光镜观察显示BBR对CNE1细胞迁移有明显的抑制作用。电镜观察结果显示,BBR抑制CNE1细胞伪足形成。结论BBR可能通过抑制Ezrin磷酸化而抑制鼻咽癌CNE1迁移。
Objective To observe the effects of berberine (BBR) on the expression of Ezrin and phos-Ezrin in nasopharyngeal carcinoma cells and its inhibitory effect on the formation and migration of pseudopods. Possible molecular mechanism of anti-NPC metastasis. Methods The nasopharyngeal carcinoma cell line (CNE1) cultured in vitro was studied. The effect of BBR on the growth of CNE1 cells was analyzed by MTT assay. The non-cytotoxic concentration (NCC) of BBR cells was used for the experiment. The expression of CNE1Ezrin and phos-Ezrin was analyzed by Western-blotting. Cell migration ability was observed with light microscope CNE1 and cell pseudopodia formation was observed with electron microscope. Transfection of Ezrin siRNA suppressed Ezrin protein expression, further confirming that BBR inhibited cell pseudopodia formation via Ezrin. Results MTT results showed that BBR at 5μM significantly inhibited the proliferation of CNE1 (P <0.05). The concentration of (0 ~ 5) μM did not significantly inhibit the proliferation of CNE1 cells. BBR significantly inhibited phos-Ezrin expression in CNE1 cells in a concentration-dependent manner in NCC. Light microscopy showed that BBR significantly inhibited the migration of CNE1 cells. Electron microscopy results showed that BBR inhibited pseudopodia formation in CNE1 cells. Conclusion BBR may inhibit the migration of CNE1 in nasopharyngeal carcinoma by inhibiting Ezrin phosphorylation.