基于高通量测序技术研究蛇床子素逆转MCF-7/Adr 细胞对阿霉素的耐药性及潜在作用机制  

Potential Mechanism of Osthole in Reversing Resistance of MCF-7/Adr Cells to Adriamycin:A Study Based on High-throughput Sequencing

作　　者：何晴 袁静静 吉章鑫 张心语 杨东梅[1] 彭辉 许娜 于庆生[2] 储俊[1] HE Qing;YUAN Jingjing;JI Zhangxin;ZHANG Xinyu;YANG Dongmei;PENG Hui;XU Na;YU Qingsheng;CHU Jun(Key Laboratory of Xin’an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Anhui Hefei 230038,China;First Department of General Surgery,The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230031,China;School of Tea and Food Science&Technology,Anhui Agricultural University,Anhui Hefei 230036,China)

机构地区：[1]安徽中医药大学新安医学教育部重点实验室,安徽合肥230038 [2]安徽中医药大学第一附属医院普外一科,安徽合肥230031 [3]安徽农业大学茶与食品科技学院,安徽合肥230036

出　　处：《安徽中医药大学学报》2025年第2期80-87,共8页Journal of Anhui University of Chinese Medicine

基　　金：安徽省高等学校自然科学研究重点项目(2023AH050835)。

摘　　要：目的基于高通量测序技术探究蛇床子素(osthole,OST)逆转MCF-7/Adr细胞对阿霉素(adriamycin,Adr)耐药性的潜在机制。方法首先采用CCK-8法检测细胞活力,采用荧光染色法评估细胞内Adr的滞留情况;其次测定多药耐药基因1(multidrug resistance 1,MDR1)和P-糖蛋白(P-glycoprotein,P-gp)水平;最后对OST给药的MCF-7/Adr细胞进行高通量测序,筛选差异表达的微小RNA(micro RNA,miRNA)并进行生物信息学分析。结果20μmol/L OST处理MCF-7/Adr细胞48 h后,对Adr的耐药倍数由20.3下降至2.8,Adr积累增加,并且MDR1 mRNA及其编码的P-gp表达水平下降。miRNA测序鉴定出92个miRNA上调,32个miRNA下调。生物信息学分析结果表明,差异表达的miRNA的主要功能是调节细胞增殖和细胞对化疗药物的耐药性;其靶基因可能与DNA结合、蛋白质运输、核苷酸结合等生物学过程相关,以及与Ras、Rap1、MAPK等信号通路有关。结论OST可逆转MCF-7/Adr细胞对Adr的耐药性,其机制可能与调节miRNA表达有关。Objective To investigate the potential mechanism of osthole(OST)in reversing the resistance of MCF-7/Adr cells to adriamycin(Adr)based on high-throughput sequencing.Methods CCK-8 assay was used to measure cell viability,and fluorescence staining was used to evaluate the retention of Adr in cells.Then the levels of multidrug resistance 1(MDR1)and P-glycoprotein(P-gp)were measured.Finally,high-throughput sequencing was performed for MCF-7/Adr cells after OST administration to identify differentially expressed micro RNA(miRNA)and perform a bioinformatics analysis.Results After MCF-7/Adr cells were treated with 20μmol/L OST for 48 hours,the drug resistance fold to Adr decreased from 20.3 to 2.8,with an increase in the accumulation of Adr,and there were reductions in the mRNA expression levels of MDR1 and P-gp encoded by MDR1.MiRNA sequencing identified 92 upregulated miRNAs and 32 downregulated miRNAs.The bioinformatics analysis showed that the differentially expressed miRNAs had the main function of regulating cell proliferation and resistance of cells to chemotherapy,and their target genes might be associated with the biological processes such as DNA binding,protein transport,and nucleotide binding and the signaling pathways such as Ras,Rap1,and MAPK.Conclusion OST can reverse the resistance of MCF-7/Adr cells to Adr,possibly by regulating the expression of miRNA.

关 键 词：MCF-7/ADR细胞 耐药性 蛇床子素 单细胞测序 微小RNA 

分 类 号：R285.5[医药卫生—中药学]