临床眼科杂志2024,Vol.32Issue(5) :444-449.DOI:10.3969/j.issn.1006-8422.2024.05.012

青藤碱抑制视网膜母细胞瘤细胞生长的作用机制研究

Study on the mechanism of Sinomenine inhibiting the growth of Retinoblastoma cells

郑虔 李翠萍 潘玉 李华林 耿红丽 朱芹 郝爽 杨淑颖
临床眼科杂志2024,Vol.32Issue(5) :444-449.DOI:10.3969/j.issn.1006-8422.2024.05.012

青藤碱抑制视网膜母细胞瘤细胞生长的作用机制研究

Study on the mechanism of Sinomenine inhibiting the growth of Retinoblastoma cells

郑虔 1李翠萍 1潘玉 1李华林 1耿红丽 1朱芹 1郝爽 1杨淑颖1
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作者信息

  • 1. 255000 淄博市妇幼保健院
  • 折叠

摘要

目的 探讨青藤碱抑制视网膜母细胞瘤系WERI-RB-1生长的作用机制.方法 采用免疫荧光法、CCK-8法和流式细胞仪分析青藤碱对WERI-RB-1细胞活力和凋亡的影响,Western blot检测相关蛋白表达.结果 不同浓度青藤碱处理的WERI-RB-1细胞中自噬相关蛋白Beclin1和LC3B表达上调,p62表达下调.WER-RBv1细胞中加入50 μmol/L青藤碱和自噬抑制剂CQ刺激24 h后,由青藤碱刺激增加的Beclin1和LC3B恢复,降低的p62蛋白也恢复;由青藤碱刺激增加的细胞自噬减少(免疫荧光法检测LC3B标记的阳性细胞);部分减轻了青藤碱诱导的细胞活力降低(CCK-8法);由青藤碱刺激增加的细胞凋亡蛋白Bax、Bcl-2蛋白恢复.青藤碱抑制PI3K/Akt/mTOR通路,激活PI3K/Akt/mTOR通路后,青藤碱对细胞生长和自噬的作用被抵消.结论 青藤碱通过体外PI3K/Akt/mTOR轴失活促进自噬来抑制视网膜母细胞瘤细胞的生长.

Abstract

Objective To explore the mechanism of Sinomenine inhibiting the growth of Retinoblastoma line WERI-RB-1.Methods Immunofluorescence,CCK-8 and flow cytometry were used to analyze the effect of Sinomenine on the viability and apoptosis of WERI-RB-1 cells,and Western blot was used to detect the expression of related proteins.Re-sults The expression of autophagy related proteins Beclin1 and LC3B was up-regulated and p62 was down regulated in WERI-RB-1 cells treated with different concentrations of Sinomenine.After 24 h stimulation with 50 μmol/L Sinomenine and autophagy inhibitor CQ,the increased Beclin1 and LC3B stimulated by Sinomenine recovered,and the decreased p62 protein also recovered;The autophagy stimulated by Sinomenine was reduced(LC3B labeled positive cells were detected by immunofluorescence);It partially alleviated the cell viability reduction induced by Sinomenine(CCK-8 method);The increased apoptosis protein Bax and Bcl-2 stimulated by Sinomenine recovered.Sinomenine inhibits the PI3K/Akt/mTOR pathway,and after activating the PI3K/Akt/mTOR pathway,the effect of Sinomenine on cell growth and autophagy is offset.Conclusions Sinomenine can inhibits retinoblastoma cell growth by promoting autophagy via inactivation of PI3K/Akt/mTOR axis in vitro.

关键词

青藤碱/视网膜母细胞瘤/作用机制

Key words

Sinomenine/retinoblastoma/mechanism

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基金项目

山东省中医药科技项目(2020Q078)

山东省医药卫生科技发展计划项目(202007020268)

出版年

2024
临床眼科杂志
安徽医科大学,安徽眼科研究所

临床眼科杂志

CSTPCD
影响因子:0.791
ISSN:1006-8422
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