黄连素通过抑制磷酸化埃兹蛋白表达阻断鼻咽癌细胞侵袭和转移的机制研究(英文)

Berberine inhibits the invasion and metastasis of nasopharyngeal carcinoma cells through Ezrin phosphorylation

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。

Objective To determine the molecular mechanism of berberine（BBR） inhibiting the metastasis and invasion of nasopharyngeal carcinoma 5-8F cells,and identify whether berberine suppresses the tumor-invasive action through inhibiting Ezrin or phosphate-Ezrin.Methods The non-cytotoxic concentration of berberine was detected by MTT assay.Filopodia formation of 5-8F cells was observed by electron microscope.The invasion and motility of 5-8F cells with berberine treatment were measured with Trans-well assay.Western blot was used to investigate the Ezrin and phos-Ezrin expression in 5-8F cells treated by berberine.pcDNA3.1-Ezrin and pcDNA3.1-Ezrin M were transfected into 6-10B cells.The inhibitory effect of berberine on the motility and invasion of 6-10B-pcDNA3.1-Ezrin and 6-10B-pcDNA3.1-Ezrin M was detected,respectively.Results Berberine non-cytotoxic concentration was 0-40 μmol/L.After being treated by berberine,filopodia of 5-8F cells obviously reduced,and the permeating artificial basement membrane cells largely decreased in both time-and concentration-dependent manner.There was significant difference compared with the control group（P0.05）.Berberine suppressed the phos-Ezrin expression of 5-8F cells in both time-and concentration-dependent manner（P0.05）,but the effect of berberine was weaker on 6-10B-pcDNA3.1-Ezrin M than on 6-10B-pcDNA3.1-Ezrin.Conclusion Berberine inhibits nasopharyngeal carcinoma cell invasion through inhibiting phos-Ezrin expression and filopodia formation.