摘要
目的:探讨盐酸川芎嗪对小鼠Lewis肺癌细胞侵袭黏附能力及上皮-间质转化的影响及其作用机制。方法:对绿色荧光蛋白(GFP)标记的小鼠Lewis肺癌细胞株(LLC-GFP)进行体外培养,经盐酸川芎嗪干预24 h后,分别采用CCK-8法检测细胞增殖抑制率,细胞黏附实验、Transwell小室实验检测细胞侵袭、黏附能力的改变,流式细胞技术、Western Blot检测E-钙黏蛋白(E-cadherin)与N-钙黏蛋白(N-cadherin)的表达变化。结果:与对照组比较,盐酸川芎嗪呈剂量依赖性抑制LLC-GFP细胞的增殖;Transwell实验显示,与对照组比较,用药组细胞的侵袭能力呈剂量依赖性下降(P<0.01);细胞黏附实验显示,与对照组黏附抑制率(14.01±1.36)%比较,应用盐酸川芎嗪需药物浓度达到2,000μg/m L时才可显著增高黏附抑制率(P<0.01);流式细胞技术及Western blot实验结果显示,与对照组比较,盐酸川芎嗪可下调N-cadherin蛋白,并上调E-cadherin蛋白的表达(P<0.01),且呈剂量依赖性。结论:盐酸川芎嗪可显著地抑制LLC-GFP细胞的生长,抑制侵袭和黏附,其机制可能与直接抑制肿瘤细胞侵袭,抑制相关黏附蛋白N-cadherin,促进黏附分子E-cadherin蛋白表达有关。
Objective： To investigate the effect of TMP on lewis lung cancer cells transition and epithelialmesenchymal conversion of mice. Methods： We cultured LLC-GFP cells in vitro, after intervention with TMP for24 h, then detected cell proliferation inhibition rate in CCK-8. Transwell assay and cell adhesion test were used to determine the inhibitory effects of TMP on cell invasion and adhesion in LLC-GFP cells. Changes of E-cadherin expression of cultured cells each group by flow cytometry were detected. Western blot assay were used to observe the expressions of EMT marker E-cadherin, N-cadherin. Results： Compared with the control group, TMP inhibited LLC-GFP cells proliferation by dose. The invasion and migration of LLC-GFP were dose-dependently inhibited in TMP group（P〈0.01）. The adhesion showed, compared with control group（14.01±1.36）, the inhibition rate of adhesion in TMP 2,000 group was dramatically increased（P〈0.01）. Flow cytometry and western blot showed that TMP can reduce N-cadherin protein and increase E-cadherin protein expression. Conclusion： TMP can inhibit the invasion and adhesion of LLC-GFP cells, and the mechanism may be related to inhibit the protein expression of N-cadherin and promote the adhesion molecule N-cadherin protein expression.
作者
迟笑怡
胡凯文
周天
CHI Xiao-yi;HU Kai-wen;ZHOU Tian(Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China)
出处
《中医药导报》
2018年第11期9-15,20,共8页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(81403250)
