贝母素乙对肝癌Huh-7细胞生长抑制作用的转录组分析  

Transcriptome Analysis of Peiminine-mediated Inhibitory Functions on Hepatocellular Carcinoma Huh-7 Cell Growth

作　　者：陈观平[1] 李静[1] 卢莺燕[1] 应栩华[1] CHEN Guanping;LI Jing;LU Yingyan;YING Xuhua(Cancer Institute of Integrated Taditional Chinese and Western Medicine,Tongde Hospital of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]浙江省立同德医院中西医结合肿瘤研究所,杭州310012

出　　处：《中国现代应用药学》2024年第17期2345-2351,共7页Chinese Journal of Modern Applied Pharmacy

基　　金：浙江省基础公益研究计划(LTGD23C040004);浙江省医药卫生科技计划项目(2021KY599)。

摘　　要：目的对经过贝母素乙处理的肝癌细胞Huh-7进行转录组测序,揭示贝母素乙对肝细胞癌的抑制机制。方法取贝母素乙组细胞,以未处理为空白对照组,应用高通量测序法检测各组表达谱,筛选出差异表达的基因,并进行生物信息学分析,实时荧光定量PCR及Western blotting验证结果。结果贝母素乙显著降低了Huh-7细胞的存活率和增殖能力,同时细胞伴有异常的形态学改变。转录组学分析显示,贝母素乙处理后,Huh-7细胞中差异表达的基因有1513个,其中680个基因表达下调,833个基因表达上调。GO和KEGG分析表明,p53、细胞衰老和细胞周期信号等经典信号通路被显著富集。实时荧光定量PCR和Western blotting结果显示差异基因和蛋白的表达趋势与转录组分析一致。结论本研究为Huh-7细胞中贝母素乙的作用机制提供了理论基础,为肝细胞癌的治疗提供了参考。OBJECTIVE To investigate the regulatory mechanisms of peiminine on human hepatocellular carcinoma by transcriptome sequencing of Huh-7 liver cancer cells treated with peiminine.METHODS Huh-7 cells treated with peiminine was selected,at the same time,untreated group was selected as control.High throughput sequencing was used to detect the expression profiles of mRNA in each group.Then the mRNA variance analysis and associated bioinformatics analysis were performed.The mRNA and protein expression level were validated by qPCR and Western blotting.RESULTS The survival rate and proliferation ability of Huh-7 cells were markedly reduced by peiminine,along with abnormal morphologic changes.The transcriptome analysis showed that 1513 genes were differentially expressed after peiminine treatment,including 680 down-regulated and 833 up-regulated genes.Gene ontology and KEGG analysis suggested that some classical pathways,such as p53,cell senescence,and cell cycle signaling,were significantly enriched.qPCR and Western blotting results showed consistent expression trends of differential genes and proteins with transcriptome analysis.CONCLUSION This study has provided a theoretical basis for regulation of peiminine in Huh-7 cells,which lays a foundation for the treatment of hepatocellular carcinoma.

关 键 词：贝母素乙 肝癌 HUH-7细胞 增殖 二代测序 

分 类 号：R285.4[医药卫生—中药学]