赤星衣酸乙酯对A549及HepG2细胞抗癌作用及机制差异研究

Study on difference in anti-cancer effect and mechanism of ethyl haemaotommate on A549 and HepG2 cells

目的:探讨赤星衣酸乙酯对A549细胞和HepG2细胞体外抗癌效果和作用机制的差异。方法:采用体外细胞培养实验,以MTT法检测赤星衣酸乙酯对A549细胞和HepG2细胞增殖的作用;应用流式细胞术,通过Annexin V/PI双染法检测赤星衣酸乙酯对细胞凋亡的影响,PI染色法检测赤星衣酸乙酯对细胞周期分布的影响;通过平板克隆形成实验观察赤星衣酸乙酯对克隆形成的影响;采用划痕实验观察赤星衣酸乙酯对细胞迁移的影响;应用mRNA转录组测序初步探讨赤星衣酸乙酯抗癌机制,采用RT-qPCR实验对测序结果进行验证。结果:赤星衣酸乙酯对A549细胞和HepG2细胞的增殖有抑制作用,并存在剂量、时间-效应关系;赤星衣酸乙酯可以抑制癌细胞克隆形成,促进癌细胞凋亡,抑制癌细胞迁移,并将大部分癌细胞阻滞于G_(0)/G_(1)期;通过调节BRCA2、IL-7R、F2发挥抗肺癌作用,通过调节STAT1、PAX8、PTGS2、LAMA1发挥抗肝癌作用。结论:赤星衣酸乙酯在A549细胞和HepG2细胞中可能通过调节不同基因导致两株细胞敏感性差异的存在。

Objective:To investigate the difference in the anti-cancer effect and mechanisms of ethyl haematommate on A549 and HepG2 cells.Methods:Cells were cultured in vitro.Proliferations of A549 and HepG2 cells were measured by MTT assay.The apoptosis was measured by Annexin V/PI.Flow cytometry was used to detect the effect of ethyl haematommate on the cell cycle.Effect of ethyl haematommate on cloning formation was explored by plate clone formation assay.Effect of ethyl haematommate on migration of A549 and HepG2 cells was detected by cell scratch test.mRNA transcriptome sequencing was used to explore the anti-cancer mechanism of ethyl haematommate,and RT-qPCR experiment was used to verify the sequencing results.Results:The results of MTT showed that the inhibition of the proliferation of A549 and HepG2 cells was a significant dose-effect relationship and time-dependent.Ethyl haematommate could inhibit the clonal formation of cancer cells,promote apoptosis,inhibit cancer cells migration,and could make most cancer cells stagnate in G_(0)/G_(1)phase.It played an anti-lung cancer by regulating BRCA2,IL-7 R and F2,and played anti-liver cancer by regulating STAT1,PAX8,PTGS2,LAMA1.Conclusion:Ethyl haematommate might regulate different genes in A549 and HepG2 cells,which led to the presence of differential sensitivity between the two cells.