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目的:探讨黄连素(berberine,BBR)抑制鼻咽癌细胞侵袭及移动的分子机制,明确BBR是否通过抑制Ezrin蛋白抑制鼻咽癌侵袭转移。方法:采用细胞增殖实验(MTT)测定BBR非毒性浓度(non-cytotoxicconcentration,NCC),扫描电子显微镜观察BBR在NCC对鼻咽癌细胞5-8F细胞伪足形成的影响,Trans-well实验检测BBR处理后鼻咽癌细胞运动侵袭能力;Western印迹检测BBR对鼻咽癌细胞Ezrin蛋白表达及其磷酸化(phos-Ezrin)的影响;基因转染技术将pcDNA3.1-Ezrin和pcDNA3.1-Ezrin M转染至6-10B细胞,分析BBR对上述细胞运动能力的影响。结果:BBR的NCC为0～40μmol/L;BBR在此浓度范围能有效地抑制细胞伪足形成,抑制鼻咽癌细胞5-8F穿过人工基底膜,与对照组比较,差异有统计学意义(P<0.05),并具有浓度依赖性和时间依赖性;BBR抑制5-8F细胞phos-Ezrin表达且呈浓度依赖性和时间依赖性,与对照组比较,差异有统计学意义(P<0.05);BBR对6-10B-pcDNA3.1-Ezrin M侵袭移动抑制作用弱于6-10B-pcDNA3.1-Ezrin。结论:BBR可以通过抑制Ezrin磷酸化表达和细胞伪足形成阻断鼻咽癌细胞的转移。
AIM: To investigate the molecular mechanism of berberine (BBR) inhibiting the invasion and metastasis of nasopharyngeal carcinoma cells and to determine whether BBR inhibits the invasion and metastasis of nasopharyngeal carcinoma by inhibiting Ezrin protein. Methods: The non-cytotoxic concentration (NCC) of BBR was determined by MTT assay. The effect of BBR on the pseudopod formation of nasopharyngeal carcinoma cell line 5-8F was observed by scanning electron microscopy. The BBR Western blotting was used to detect the effect of BBR on Ezrin protein expression and phosphorylation (Phos-Ezrin) expression in nasopharyngeal carcinoma cells. Gene transfection technology pcDNA3.1-Ezrin and pcDNA3.1-Ezrin M transfection to 6-10B cells, BBR analysis of the above cell motility. Results: The NCC of BBR ranged from 0 to 40 μmol / L. BBR in this concentration range could effectively inhibit the formation of pseudopodia and inhibit 5-8F of nasopharyngeal carcinoma cells crossing the artificial basement membrane, which was significantly different from the control group (P <0.05). BBR inhibited the expression of phos-Ezrin in 5-8F cells in a concentration-dependent manner and in a time-dependent manner. Compared with the control group, the difference was statistically significant (P <0.05 ). The inhibitory effect of BBR on the invasion of 6-10B-pcDNA3.1-Ezrin M was weaker than that of 6-10B-pcDNA3.1-Ezrin. CONCLUSION: BBR can block the metastasis of nasopharyngeal carcinoma cells by inhibiting Ezrin phosphorylation and cell pseudopodia formation.