Li Ming,Li Yong,Huang Zhao,et al.Effect of Serabelisib on human hepatoblastoma HepG2 cells and its mechanisms[J].Journal of Clinical Pediatric Surgery,2020,19(05):410-414.[doi:10.3969/j.issn.1671-6353.2020.05.007]
Serabelisib抗人肝母细胞瘤HepG2细胞的作用及其机制探讨
- Title:
- Effect of Serabelisib on human hepatoblastoma HepG2 cells and its mechanisms
- Keywords:
- Liver Neoplasms; PI3K; Serabelisib; HepG2; Cell Proliferation; Apoptosis
- 分类号:
- R726;R735.7
- 摘要:
- 目的 探讨小分子抑制剂Serabelisib抑制PI3K-Akt信号通路活性及对人肝母细胞瘤细胞系增殖、凋亡的效应作用及其相关机制。方法 采用不同浓度的小分子抑制剂Serabelisib（0 μmol/L、2 μmol/L、4 μmol/L、8 μmol/L）处理人肝母细胞瘤细胞HepG2细胞，通过四氮唑盐比色法检测Serabelisib对细胞增殖的抑制作用，采用流式细胞仪检测细胞凋亡变化情况，采用蛋白质免疫印迹检测Serabelisib处理后HepG2细胞中PI3K信号通路活性及凋亡相关蛋白表达变化情况。结果 当Serabelisib浓度≥4 μmol/L时，对人肝母细胞瘤细胞系HepG2细胞增殖具有显著抑制作用，随着Serabelisib浓度增加，其对HepG2细胞增殖的抑制率逐渐增大，具有剂量和时间依赖性。随着Serabelisib浓度增加，HepG2细胞凋亡率明显升高，同样具有剂量依赖性。蛋白质免疫印迹检测发现Serabelisib处理后的HepG2细胞中PI3K信号通路中关键激酶Akt磷酸化水平降低，促凋亡蛋白PARP 89KD裂解片段表达增加。结论 Serabelisib通过抑制PI3K/Akt信号转导通路，抑制人肝母细胞瘤HepG2细胞增殖，并诱导凋亡。
- Abstract:
- Objective To explore the inhibitory effect of small molecule inhibitor Serabelisib on the activity of PI3K Akt signaling pathway and its mechanism on the proliferation and apoptosis of human hepatoblastoma cell line.Methods Different concentrations of Serabelisib (0,2,4,8 μmol/L) were employed for treating cells.The inhibition of cellular proliferation was detected by methyl thiazolyl tetrazolium (MTT).The changes of apoptosis were detected by flow cytometry.The cellular activity of PI3K signaling pathway and the cellular expression of apoptosis-related proteins were detected by Western blot.Results A concentration of Serabelisib ≥4 μmol/L had a significant inhibitory effect on cellular proliferation.With a rising concentration of Serabelisib,its inhibitory rate on cellular proliferation gradually increased in a dose/time-dependent manner while cellular apoptotic rate rose significantly in a dose-dependent manner.Western blot indicated that the phosphorylation of Akt,a key kinase in PI3K signaling pathway,decreased and the expression of PARP 89kd fragment spiked in Serabelisib-treated HepG2 cells.Conclusion Serabelisib can inhibit the proliferation and apoptosis by inhibiting PI3K/Akt signal transduction pathway in HepG2 cells.
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备注/Memo
收稿日期:2020-03-18。
基金项目:湖南省卫生健康委课题（编号：C2019021）；湖南省出生缺陷协同防治科技重大专项（编号：2019SK1010）
作者简介:黎明,Email:dawn1212@sina.com