摘要
目的探究miR-27b-3p在口腔鳞癌(oral squamous cell carcinoma,OSCC)耐药细胞增殖、凋亡和顺铂(cisplatin,DDP)耐药性中的作用及潜在的机制.方法RT-qPCR检测miR-27b-3p和HOXB8 mR-NA的表达;Western印迹分析HOXB8及耐药蛋白MRP1的表达;CCK-8分析细胞活力,克隆形成实验分析细胞增殖,流式细胞仪检测细胞凋亡.miR-27b-3p和HOXB8之间的相互作用通过在线软件Targetscan预测并通过双荧光素酶报告基因分析证实.裸鼠异种移植模型测试miR-27b-3p在体内OSCC DDP耐药中的作用.结果miR-27b-3p在DDP耐药OSCC组织和OSCC细胞系中明显低表达,且在DDP耐药OSCC细胞系(CAL-27/DDP)中的表达明显低于正常OSCC细胞系(CAL-27)(P<0.05);而HOXB8 mRNA在DDP耐药OSCC组织和OSCC细胞系中明显高表达,在DDP耐药OSCC细胞系(CAL-27/DDP)中的表达明显高于正常OSCC细胞系(CAL-27)(P<0.05).miR-27b-3p模拟物可明显抑制CAL-27/DDP细胞活力、克隆形成能力和耐药蛋白MRP1的表达,促进细胞凋亡(P<0.05);而HOXB8过表达可部分逆转miR-27b-3p模拟物对CAL-27/DDP细胞增殖、凋亡以及耐药性的影响(P<0.05).HOXB8与miR-27b-3p存在靶向调控作用.瘤内注射miR-27b-3p agomir显著缓解了体内异种移植耐药细胞的生长,同时降低了HOXB8和MRP1的表达(P<0.05).结论miR-27b-3p可能通过靶向HOXB8调节OSCC耐药细胞的增殖、凋亡和DDP耐药性.
Objective To explore the role and potential mechanism of miR-27b-3p in the pro-liferation,apoptosis and cisplatin(DDP)resistance of oral squamous cell carcinoma(OSCC)re-sistant cells.Methods RT-qPCR was applied to detect the expression of miR-27b-3p and HOXB8 mRNA.Western blotting was applied to detect the expression of HOXB8 and drug resistance protein MRP1.CCK-8 assay was conducted to measure the cell viability,colony formation assay was used to analyze cell proliferation,and flow cytometry to detect apoptosis.The interaction between miR-27b-3p and HOXB8 was predicted by the online software Targetscan and confirmed by dual-luciferase re-porter gene assay.A nude mouse xenograft model was used to test the role of miR-27b-3p in OSCC DDP resistance in vivo.Results The expression level of miR-27b-3p was obviously lower in DDP-resistant OSCC tissues and OSCC cell lines,and the expression level in DDP-resistant OSCC cell lines(CAL-27/DDP)was obviously lower than that in normal OSCC cell lines(CAL-27)(P<0.05).HOXB8 mRNA,by contrast,was highly expressed in DDP-resistant OSCC tissues and OS-CC cell lines,and the expression in DDP-resistant OSCC cell line(CAL-27/DDP)was obviously higher than that in normal OSCC cell line(CAL-27)(P<0.05).miR-27b-3p mimic could obviously inhibit the cell viability,colony formation ability and the expression of drug resistance protein MRP1 in CAL-27/DDP cells,and promote the cell apoptosis(P<0.05).Overexpression of HOXB8 could partially reverse the effects of miR-27b-3p mimics on the proliferation,apoptosis and drug resistance in CAL-27/DDP cells(P<0.05).HOXB8 had a targeted regulatory role with miR-27b-3p.Intratumoral injection of miR-27b-3p agomir obviously alleviated the growth of resistant cells in the xenograft in vivo,and reduce the expression level of HOXB8 and MRP1(P<0.05).Conclusion miR-27b-3p may regulate the proliferation,apoptosis and DDP resistance of OSCC drugresistant cells by targeting HOXB8.
作者
周金阔
史晓晶
刘倩峰
刘广顺
姜磊
张晋弘
ZHOU Jinkuo;SHI Xiaojing;LIU Qianfeng;LIU Guangshun;JIANG Lei;ZHANG Jinhong(The Second Department of Stomatology,the First Hospital of Hebei Medical University,Shijiazhuang,050000,China;the First Department of Stomatology,the First Hospital of Hebei Medical University,Shijiazhuang,050000,China;Central Laboratory,the First Hospital of Hebei Medical University,Shijiazhuang,050000,China)
出处
《医学分子生物学杂志》
CAS
2023年第3期202-208,共7页
Journal of Medical Molecular Biology
基金
河北省2022年度医科科学研究课题(No.20221387)。
