米诺地尔在肝癌中的作用研究

Study on the role of minoxidil in hepatocellular carcinoma

目的初步探讨米诺地尔对肝癌细胞增殖、迁移和侵袭的影响及其可能分子机制。方法通过CCK-8法、划痕实验、Transwell实验和流式细胞分析分别检测米诺地尔处理人肝癌细胞系Huh7和MHCC97H后对细胞增殖、迁移、侵袭和凋亡的影响;通过蛋白质组定量实验分析米诺地尔处理后肿瘤细胞蛋白表达的差异,并对差异蛋白进行生物学进程、细胞组成和分子功能的富集分析,以及基于KEGG数据库注释的通路分析。结果米诺地尔可抑制Huh7和MHCC97H细胞的增殖、迁移和侵袭,但不影响凋亡;米诺地尔处理MHCC97H细胞48 h后共鉴定到279种差异蛋白,其中上调蛋白89种,下调蛋白190种。差异蛋白主要涉及细胞黏附、氧化还原、DNA复制等生物学过程,以及真核生物核糖体生物发生、类固醇生物合成、DNA复制等信号通路。不仅胞内蛋白,胞外蛋白表达也受到影响。结论米诺地尔可在体外抑制肝癌细胞的增殖、迁移和侵袭。蛋白质组分析结果表明,米诺地尔可能通过影响与肿瘤增殖、迁移和侵袭相关的重要蛋白及通路,在肝细胞癌中发挥抗肿瘤作用。

Objective To explore the effect of minoxidil on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells and the possible molecular mechanism.Methods The effects of minoxidil on cell proliferation,migration,invasion and apoptosis were detected by CCK-8 assay,wound-healing assay,Transwell assay and flow cytometry after minoxidil treatment of human HCC cell lines Huh7 and MHCC97H.Quantitative proteome profiling was performed to analyze the protein expression differences in cancer cells after minoxidil treatment.And the properties of differentially expressed proteins were revealed by enrichment analysis of the biological process,cellular component and molecular function.Pathway alterations were also analyzed based on KEGG database annotations.Results Minoxidil inhibited the proliferation,migration and invasion of HCC cells,without significant effect on apoptosis.There were 279 differentially expressed proteins identified in MHCC97H cells after minoxidil treatment.Among them,89 proteins were up-regulated and190 were down-regulated.The differentially expressed proteins were mainly involved in such biological processes as cell-cell adhesion,oxidation-reduction processes,DNA replication,and signaling pathways including ribosome biogenesis in eukaryotes,steroid biosynthesis and DNA replication.The expressions of both intracellular and extracellular proteins were affected.Conclusion Minoxidil can inhibit the proliferation,migration and invasion of HCC cells in vitro.Proteomic analysis showed that minoxidil might play an anti-tumor role in HCC by affecting the important proteins and pathways related to tumor proliferation,migration and invasion.