基于RNA-seq探讨二氢丹参酮Ⅰ抑制胃癌细胞增殖的作用机制

Mechanism of RNA-seq-based dihydrotanshinone Ⅰ in gastric cancer cell proliferation inhibition

目的为了阐明二氢丹参酮Ⅰ(DHTS)抗胃癌的作用机制,为其在临床应用中提供思路及数据支持。方法采用RNA测序(RNA-seq)技术探索DHTS抗胃癌的具体作用机制。结果发现4353个差异基因表达。这些差异基因参与了肿瘤发生发展的多条信号通路,主要涉及PI3K/Akt/m TOR信号转导通路等。在测序结果中,有89个差异基因被注释到PI3K/Akt/m TOR信号转导通路,推测该信号转导通路和DHTS诱导胃癌细胞凋亡密切相关。Western-blot检测验证了DHTS可显著抑制PI3K、Akt及m TOR的磷酸化,且显著增加Caspase-3和LC3蛋白的表达水平,从而促进胃癌细胞凋亡和自噬,抑制胃癌细胞增殖。结论DHTS促进胃癌细胞凋亡,是通过活性氧介导的氧化应激促进细胞凋亡,具体作用机制可能是:活性氧介导的氧化应激抑制PI3K/Akt/m TOR信号通路,从而诱导细胞凋亡以及自噬,进而抑制胃癌细胞的增殖生长。

Objective To elucidate the mechanism of dihydrotanshinone Ⅰ(DHTS)against gastric cancer and to provide ideas and data support for its clinical treatment and application.Methods RNA sequencing was used to explore the specific mechanism of DHTS against gastric cancer.Results It was found that 4353 differential genes were expressed,which were involved in multiple signaling pathways in tumor development,mainly involving the PI3K/Akt/mTOR signal transduction pathway.In the sequencing results,89 differential genes were annotated to the PI3K/Akt/mTOR signal transduction pathway,which was speculated to be closely related to DHTS-induced apoptosis in gastric cancer cells.Western-blot analysis verified that DHTS could markedly inhibit the phosphorylation of PI3K,Akt,and mTOR,and increase the expression levels of caspase-3 and LC3 proteins,thereby promoting gastric cancer cell apoptosis and autophagy and inhibiting gastric cancer cell proliferation.Conclusion DHTS promotes apoptosis of gastric cancer cells through reactive oxygen-mediated oxidative stress.The specific mechanism may be that reactive oxygen-mediated oxidative stress inhibits the PI3K/Akt/mTOR signaling pathway,thereby inducing apoptosis and autophagy,and then inhibiting the proliferation and growth of gastric cancer cells.