摘要
目的:探讨乳积方对乳腺癌MDA-MB-231细胞miRNA表达谱的影响。方法:将体外培养的MDA-MB-231细胞随机分为正常对照组和乳积方处理组,培养后使用miRNA芯片技术检测乳积方对MDA-MB-231细胞miRNA表达谱的影响。采用mirdbV5和TaregetScan7.1数据库对差异表达miRNA进行靶基因预测,并运用Go&Pathway分析获取其相关的生物学功能和信号通路。RT-PCR用于验证miRNA表达谱结果。结果:乳积方药物作用MDA-MB-231细胞72h后,miRNA芯片检测筛选出表达差异大于2倍的基因共有219个,其中上调109个,下调110个。进一步增大筛选标准(Fore-Ground强),miRNA的表达谱存在25个差异表达的miRNA,这25个差异表达miRNA靶基因显著富集于PI3K-Akt通路、miRNA癌症通路等信号通路,涉及细胞增殖、凋亡和迁移等生物过程。从这25个差异表达的miRNA中,选取21个miRNA进行RT-PCR验证,除hsa-miR-601外,其他20个miRNA的QPCR结果与芯片结果一致。结论:乳积方可能参与乳腺癌相关miRNA的表达调控,进而影响相关信号通路的转导,从而发挥其抗肿瘤作用。
Objective:To investigate the effects of Ruji Formula on miRNA expression of MDA-MB-231 cells in the treatment of breast cancer.Methods:MDA-MB-231 cells cultured in vitro were randomly divided into the normal control group and the Ruji Formula intervention group.miRNA chip technology was used to detect the effect of Ruji Formula on the miRNA expression of MDA-MB-231 cells after being cultured.MirdbV5 and TaregetScan7.1 databases were used to predict the target genes of differentially expressed miRNA,and Go&Pathway analysis was used to obtain their relevant biological functions and signaling pathways.Quantitative real-time RT-PCR was used to verify miRNA expression profile results.Results:After 72 hours of MDA-MB-231 cells intervened with Ruji Formula,a total of 219 genes with expression difference more than two times were screened out by miRNA chip detection,among which 109 were up-regulated and 110 were down-regulated.Further enhancing the selection criteria,there were 25 differentially expressed miRNA,which were significantly enriched in signaling pathways of PI3K-Akt and the miRNA cancer pathway,involving biological processes,such as cell proliferation,apoptosis and migration.From the 25 differentially expressed miRNA,21 miRNA were selected for quantitative real-time RT-PCR verification.Except hsa-miR-601,QPCR results of the other 20 miRNA were consistent with chip results.Conclusion:Ruji Formula may be involved in regulating expression of miRNA related to breast cancer,thus to affect the transduction of relevant signaling pathways and exert its anti-tumor effect.
作者
李宏良
田华琴
王斌
李岩
王凯
梁贵文
陈学彰
朱志霞
陈锡康
LI Hongliang;TIAN Huaqin;WANG Bin;LI Yan;WANG Kai;LIANG Guiwen;CHEN Xuezhang;ZHU Zhixia;CHEN Xikang(Oncology Center, Foshan Hospital of TCM Affiliated to Guangzhou University of Chinese Medicine, Foshan 528000, China;Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;Guangdong Bio-Healtech Advanced Co., Ltd., Foshan 528300, China)
出处
《中医药学报》
CAS
2022年第6期32-39,共8页
Acta Chinese Medicine and Pharmacology
基金
广东省中医药局科研课题重点项目(20173015)。
作者简介
李宏良(1972-),男,博士,主任中医师,研究方向:中西医结合治疗肿瘤血液病临床及实验研究;通讯作者:田华琴(1963-),女,教授,主任中医师,研究方向:中医药防治乳腺癌术后复发转移。
