CAJ | 학술논문

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-cytotoxic concentration,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0~40 μmol/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.
목적:탐토황련소(berberine,BBR)억제비인암세포침습급이동적분자궤제,명학BBR시부통과억제Ezrin단백억제비인암침습전이.방법:채용세포증식실험(MTT)측정BBR비독성농도(non-cytotoxic concentration,NCC),소묘전자현미경관찰BBR재NCC대비인암세포5-8F세포위족형성적영향,Trans-well실험검측BBR처리후비인암세포운동침습능력;Western인적검측BBR대비인암세포Ezrin단백표체급기린산화(phos-Ezrin)적영향; 기인전염기술장pcDNA3.1-Ezrin화pcDNA3.1-Ezrin M전염지6-10B 세포,분석BBR대상술세포운동능력적영향.결과:BBR적NCC위0~40 μmol/L;BBR재차농도범위능유효지억제세포위족형성,억제비인암세포5-8F천과인공기저막,여대조조비교,차이유통계학의의(P<0.05),병구유농도의뢰성화시간의뢰성;BBR억제5-8F 세포phos-Ezrin표체차정농도의뢰성화시간의뢰성,여대조조비교,차이유통계학의의(P<0.05); BBR 대6-10B-pcDNA3.1-Ezrin M 침습이동억제작용약우6-10B-pcDNA3.1-Ezrin.결론:BBR가이통과억제Ezrin린산화표체화세포위족형성조단비인암세포적전이.
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 (P<0.05). Berberine suppressed the phos-Ezrin expression of 5-8F cells in both time- and concentration-dependent manner (P<0.05), 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.