陕西医学杂志2024，Vol.53Issue(6)：754-758,763,6.DOI:10.3969/j.issn.1000-7377.2024.06.007
基于长链非编码RNA HOX转录反义RNA探讨次乌头原碱对人肝癌SK-Hep-1细胞自噬的影响实验研究
Effect of protopine on autophagy in human liver cancer SK-Hep-1 cells based on lncRNA HOTAIR
摘要
Abstract
Objective:To investigate the effect of protopine on autophagy in human liver cancer SK-Hep-1 cells based on long non-coding RNA(lncRNA)HOX transcript antisense RNA(HOTAIR).Methods:Logarithmic phase human hepatoma SK-Hep-1 cells were taken,and lncRNA HOTAIR suppression plasmid lncRNA HOTAIR-shRNA was transfected into SK-Hep-1 cells,which were randomly divided into transfection group and combination group.The transfection group was cultured routinely,and the combination group was added with protopine(final concentra-tion 200 μmol/L).Another SK-Hep-1 cells were randomly divided into blank group and protopine group,and the blank group was cultured routinely,and the protopine group was added with protopine(final concentration 200 μmol/L).The cells were collected after 48 hours,and RT-qPCR was used to detect the expression of lncRNA HOTAIR in each group;flow cytometry was used to detect the apoptosis rate of each group;GFP-LC3 transfection was used to detect the autophagy of each group;immunoblotting was used to detect the expression of Beclin1,P62,LC3 Ⅰ protein in cells.Results:Compared with the blank group,the expression of lncRNA HOTAIR in the protopine group was in-creased,and the expression of lncRNA HOTAIR in the transfection group was decreased(all P＜0.05).Compared with the protopine group,the expression of lncRNA HOTAIR in the combination group was decreased(P＜0.05).Compared with the transfection group,the expression of lncRNA HOTAIR in the combination group was increased(P＜0.05).Compared with the blank group,the apoptosis rate and the expression of Beclin1,P62,LC3 Ⅰ protein in the protopine group was increased,and the number of GFP-LC3 dots was increased(all P＜0.05);the apoptosis rate and the expression of Beclin1,P62,LC3 Ⅰ protein in the transfection group were decreased,and the number of GFP-LC3 dots was decreased(all P＜0.05).Compared with the protopine group,the apoptosis rate and the expression of Bec-lin1,P62,LC3 Ⅰ protein in the combination group were decreased,and the number of GFP-LC3 dots was decreased(all P＜0.05).Compared with the transfection group,the apoptosis rate and the expression of Beclin1,P62,LC3 Ⅰprotein in the combination group were increased,and the number of GFP-LC3 dots was increased(all P＜0.05).Conclusion:Protopine may induce excessive autophagy and promote apoptosis of human hepatoma SK-Hep-1 cells,and its mechanism may be related to the up-regulation of lncRNA HOTAIR expression.关键词
肝癌/次乌头原碱/长链非编码RNA/HOX转录反义RNA/自噬/细胞凋亡Key words
Hepatoma/Protopine/Long non-coding RNA/HOX antisense RNA/Autophagy/Apoptosis分类
医药卫生引用本文复制引用
濮忠建,杨军,杨帆,王亚军..基于长链非编码RNA HOX转录反义RNA探讨次乌头原碱对人肝癌SK-Hep-1细胞自噬的影响实验研究[J].陕西医学杂志,2024,53(6):754-758,763,6.基金项目
江苏省中医药科技发展计划项目(YB2020070) （YB2020070）
江苏省南通市中医医疗联盟科研课题(TZYK202218) （TZYK202218）
