基于转录组测序分析人参皂苷Rh2抗宫颈癌的潜在作用机制

Study on the Potential Mechanism of Ginsenoside Rh2 Against Cervical Cancer Based on Transcriptome Sequencing

目的研究人参皂苷Rh2抗宫颈癌潜在作用机制。方法采用CCK-8法检测人参皂苷Rh2处理后的细胞活力。采用转录组测序技术检测对照组及人参皂苷Rh2处理组样本生物学重复的有效性、筛选差异表达基因、探寻人参皂苷Rh2抗宫颈癌相关的靶基因和信号通路。qPCR验证转录组数据可靠性。采用分光光度法检测Caspase 1/4酶活性。通过TUNEL染色检测人参皂苷Rh2诱导宫颈癌细胞死亡的形式。结果人参皂苷Rh2的IC_(50)值为45μmol/L。各组测序样本生物学重复相似度高。GO和KEGG分析结果揭示人参皂苷Rh2主要参与免疫系统进程、细胞增殖、转录调节活性等功能以及细胞焦亡相关NOD样受体信号通路。qPCR验证显示细胞焦亡相关差异表达基因的表达趋势与转录组数据一致。人参皂苷Rh2干预后HeLa细胞的Caspase 1/4酶活性增加,细胞焦亡水平明显升高。结论人参皂苷Rh2可能通过激活NOD样受体信号通路中的GSDMD、Caspase 1、Caspase 4、NLRP3、IL-1β基因表达诱导细胞焦亡发挥抗宫颈癌作用。

Objective To study the potential mechanism of ginsenoside Rh2 anti-cervical cancer based on transcriptome sequencing technology.Methods The cell viability after ginsenoside Rh2 treatment was detected by CCK-8 method.Transcriptome sequencing technology was used to verify the validity of biological replication of samples in the control group and ginsenoside Rh2-treated group,screen differentially expressed genes,and explore the target genes and signaling pathways related to ginsenoside Rh2 anti-cervical cancer.qPCR validates the reliability of transcriptome data.The enzyme activity of Caspase 1/4 was detected by spectrophotometry.The form of ginsenoside Rh2-induced cervical cancer cell death was detected by TUNEL staining.Results The IC_(50) value of ginsenoside Rh2 was 45μmol/L.The biological replicates of each group were highly similar.GO and KEGG analysis showed that ginsenoside Rh2 was mainly involved in immune system process,cell proliferation,transcriptional regulation and other functions,and NOD-like receptor signaling pathway related to cell pyroptosis.qPCR validation showed that the expression trends of pyroptosis-related differentially expressed genes were consistent with transcriptome data.After the intervention of ginsenoside Rh2,the activity of Caspase 1/4 enzymes in HeLa cells increased,and the level of pyroptosis was significantly increased.Conclusion Ginsenoside Rh2 may induce pyroptosis by activating GSDMD,Caspase I,Caspase4,NLRP3,IL-1βgene expression in NOD-like receptor signaling pathway to exert anti-cervical cancer effect.