基于转录组学探讨右旋龙脑对顺铂耐药非小细胞肺癌的增敏作用及机制  

Sensitizing effect of d-borneol on cisplatin-resistant NSCLC based on transcriptomics and its mechanism

作　　者：李金秀 王佳俊 马荣 谢倩[1,2] 龚道银[3] 王建[1,2] 曾南[1,2] LI Jin-xiu;WANG Jia-jun;MA Rong;XIE Qian;GONG Dao-yin;WANG Jian;ZENG Nan(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Key Laboratory of Systematic Research of Distinctive Chinese Medicine Resources in Southwest China,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China)

机构地区：[1]成都中医药大学药学院,四川成都611137 [2]成都中医药大学西南特色中药资源重点实验室,四川成都611137 [3]成都中医药大学附属医院,四川成都610075

出　　处：《中国药理学通报》2023年第6期1105-1114,共10页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 81873023)。

摘　　要：目的采用转录组测序(RNA-sequencing,RNA-seq)技术挖掘右旋龙脑联合顺铂对顺铂耐药的非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞的作用关键靶点,并验证其机制。方法将顺铂耐药的人大细胞肺癌细胞(H460/CDDP)接种于4周龄雄性BALB/c裸鼠右侧腋下,构建异种移植肿瘤模型,将其分为对照组、溶剂组、顺铂组、联用组(右旋龙脑+顺铂),每组6只,药物干预14 d。末次给药24 h后,取肿瘤组织,转录组测序;实时反转录聚合酶链反应(RT-PCR)和免疫组化法(IHC)检测细胞周期相关分子表达。结果顺铂组与联用组在基因表达水平存在差异,且明显富集于细胞周期。RT-PCR和IHC结果显示,右旋龙脑联合顺铂组能明显抑制细胞周期蛋白(CCNA2,CCND3)和细胞周期蛋白依赖性激酶(CDK2,CDK6)的表达量,并促进其上游分子细胞周期蛋白依赖性激酶抑制剂CDKI(P21,P27)的表达(P<0.05,P<0.01)。结论右旋龙脑联合顺铂通过上调P21、P27表达,抑制cyclinA2/D3及CDK2/6表达,诱导细胞周期阻滞,抑制H460/CDDP细胞的恶性增殖,进而达到抗耐药增敏的作用。Aim To explore the key targets of d-borneol combined with cisplatin for sensitization of cisplatin-resistant NCSLC cells by RNA-Seq and verify its mechanism.Methods Cisplatin-resistant human large cell lung cancer cells(H460/CDDP)were inoculated into the right armpit of male BALB/c nude mice(4 weeks old)to construct a xenograft tumor model.Then they were randomly divided into control group,vehicle group,cisplatin group,and combination group(d-borneol+cisplatin)with 6 nude mice and treated for 14 d.After last administration of 24 h,the tumor tissue was taken for RNA-Seq.And then real-time reverse transcription polymerase chain reaction(RT-PCR)and immunohistochemistry(IHC)were used to verify the expression of cell cycle-related molecules.Results RNA-seq analysis showed that there were significant differences in gene expression between the cisplatin group and combined group,and they were significantly enriched in cell cycle.RT-PCR and IHC results showed that d-borneol combined with cisplatin could significantly inhibit the expressions of cyclins(cyclin A2,cyclin D3)and cyclin-dependent kinases(CDK2,CDK6)and promote the expression of its upstream molecular cyclin-dependent kinase inhibitor CDKI(P21,P27)(P<0.05,P<0.01).Conclusions d-Borneol increases the sensitivity of cisplatin by increasing the expression of P21 and P27 and inhibiting the expression of cyclinA2/D3 and CDK2/6 to induce cell cycle arrest and inhibit the malignant proliferation of H460/CDDP cells,thereby achieving the effect of anti-drug sensitization.

关 键 词：右旋龙脑 顺铂 肺癌 耐药 RNA测序 细胞周期 

分 类 号：R-332[医药卫生] R284.1R329.28R394.2R734.2R979.19