周蓉蓉 高鹏飞 张文意 张艳聪 马维维 刘秋雨 史新元 连增林

摘要：目的 观察槐耳板蓝根双向发酵产物对乳腺癌MCF-7细胞迁移、侵袭和相关因子的影响，探讨其相关机制。方法 以人乳腺癌MCF-7细胞为模型，实验分为对照组、板蓝根组、槐耳组和槐耳板蓝根组。MTT法检测细胞增殖；细胞划痕实验检测MCF-7细胞迁移能力；Transwell细胞侵袭实验检测MCF-7细胞侵袭能力；细胞黏附实验检测MCF-7细胞黏附能力；半定量RT-PCR检测MCF-7细胞基质金属蛋白酶（MMP-9）和波形蛋白（Vimentin）基因表达。结果 与板蓝根组和槐耳组比较，槐耳板蓝根组明显抑制MCF-7细胞增殖、迁移、侵袭和黏附（P<0.05），抑制MMP-9和Vimentin的基因表达（P<0.05）。结论 槐耳板蓝根双向发酵提取物可能通过下调MMP-9和Vimentin的基因表达抑制乳腺癌MCF-7细胞迁移、侵袭。

关键词：槐耳；板蓝根；双向发酵；乳腺癌；迁移；侵袭；基质金属蛋白酶；波形蛋白

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）05-0064-05

Effects of Trametes Robiniophila Murr and Isatidis Radix Bidirectional Fermentation Products on Migration/Invasion of Human Breast Cancer MCF-7 Cells and Relevant Factors ZHOU Rong-rong1， GAO Peng-fei1， ZHANG Wen-yi1， ZHANG Yan-cong1， MA Wei-wei1， LIU Qiu-yu1， SHI Xin-yuan1， LIAN Zeng-lin2 （1. School of Pharmaceutical Sciences， Beijing University of Chinese Medicine， Beijing 100102， China； 2. Beijing Handian Academy of Chinese Medicine， Handian Group， Beijing 100103， China）

Abstract： Objective To observe the effects of Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on the migration and invasion of human breast cancer MCF-7 cells and relevant factors； To discuss relevant mechanism of action. Methods Breast carcinoma cell line MCF-7 was used as research subject in this experiment. Control group， Isatidis Radix group， Trametes robiniophila Murr group， and Trametes robiniophila Murr and Isatidis Radix group were included in the experiment. The effects of Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on MCF-7 cell proliferation were measured by MTT method. Cell scratch assay， transwell assay and adhesion assay were used to measure the effects of Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on the migration， invasion and adhesion capability of MCF-7 cells， respectively. The effects of Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on the mRNA expression of MMP-9 and Vimentin were measured by RT-PCR. Results Compared with Isatidis Radix group and Trametes robiniophila Murr group， Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products could significantly inhibit the proliferation， migration， invasion and adhesion capability of MCF-7 cells （P<0.05）. Similarly， Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products reduced the mRNA expression of MMP-9 and Vimentin （P<0.05）. Conclusion Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products may down-regulate the expression of MMP-9 and Vimentin to inhibit the migration， invasion and adhesion capabilities of MCF-7 cells.

Key words： Trametes robiniophila Murr； Isatidis Radix； bidirectional fermentation； breast cancer； migration； invasion； MMP-9； Vimentin

基金项目：北京市科技新星计划交叉学科项目（xxhz201210）

通讯作者：史新元，E-mail：xyshi@126.com；连增林，E-mail：zenglinlian@aliyun.com

槐耳板蓝根双向发酵产物是基于20世纪80年代庄毅教授创立的现代中药新型双向发酵技术研制而成[1]。槐耳菌适应性广泛，在发酵过程中可产生丰富的次生代谢产物，具有较好的辅助抗肿瘤作用[2-5]。板蓝根含有丰富的营养物质，其化学成分基础研究已较为成熟，且其所含化学成分靛玉红已被证实具有提高免疫及抗肿瘤作用[6]。故采用具有一定活性成分的板蓝根作为药性基质，将槐耳和板蓝根作为一对组合开展双向发酵研究，以期发生增强药效的协同作用，获得更好的生物学效应。本课题组前期研究表明，发酵过程中菌质（靛蓝、靛玉红、精氨酸为主成分）薄层斑点随发酵时间呈现规律变化，随着发酵时间的延长，有新的斑点出现[7]。……