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

关键词:  贝母素乙  肝癌  Huh-7  增殖  二代测序

Key words:  peiminine  HCC  Huh-7 cells  cell growth  RNA-seq

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HCC细胞的抑制机制。方法 取贝母素乙处理组细胞，以未处理组对照组，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WB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。qPCR和WB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HCC的治疗提供了一定的素材。

Abstract:

OBJECTIVE To investigate the regulatory mechanisms of peiminine on human HCC Huh-7 cells. 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 WB. 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 WB 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 HCC.