[1]濮忠建,杨 军,杨 帆,等.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,(6):754-758,763.[doi:DOI:10.3969/j.issn.1000-7377.2024.06.007]
　PU Zhongjian,YANG Jun,YANG Fan,et al.Effect of protopine on autophagy in human liver cancer SK-Hep-1 cells based on lncRNA HOTAIR[J].,2024,(6):754-758,763.[doi:DOI:10.3969/j.issn.1000-7377.2024.06.007]
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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
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年6期
页码:
754-758,763
栏目:
基础研究
出版日期:
2024-06-05

文章信息/Info

Title:
Effect of protopine on autophagy in human liver cancer SK-Hep-1 cells based on lncRNA HOTAIR
作者:
濮忠建; 杨 军; 杨 帆; 王亚军
(海安市中医院肿瘤科,江苏 海安 226600)
Author(s):
PU Zhongjian; YANG Jun; YANG Fan; WANG Yajun
(Department of Oncology,Haian Hospital of Traditional Chinese Medicine,Haian 226600,China)
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
Keywords:
Hepatoma;  Protopine;  Long non-coding RNA;  HOX antisense RNA;  Autophagy;  Apoptosis
分类号:
R 285.5
DOI:
DOI:10.3969/j.issn.1000-7377.2024.06.007
文献标志码:
A
摘要:
目的:基于长链非编码RNA(lncRNA)HOX转录反义RNA(HOTAIR)探讨次乌头原碱对人肝癌SK-Hep-1细胞自噬的影响。方法:取对数期人肝癌SK-Hep-1细胞,将lncRNA HOTAIR抑制质粒lncRNA HOTAIR-shRNA转染至SK-Hep-1细胞,随机分为转染组和联合组,转染组常规培养,联合组加入次乌头原碱(终浓度200 μmol/L)。另取SK-Hep-1细胞随机分为空白组和次乌头原碱组,空白组常规培养,次乌头原碱组加入次乌头原碱(终浓度200 μmol/L)。48 h后收集细胞,实时荧光定量PCR(RT-qPCR)检测各组lncRNA HOTAIR表达量; 流式细胞仪检测各组凋亡率; 绿色荧光蛋白(GFP)-微管相关蛋白1轻链3(LC3)转染检测各组自噬情况; 免疫印迹法检测细胞Beclin1、P62、微管相关蛋白1轻链3 Ⅰ(LC3 Ⅰ)蛋白相对表达量。结果:与空白组比较,次乌头原碱组lncRNA HOTAIR表达量升高,转染组lncRNA HOTAIR表达量降低(均P<0.05)。与次乌头原碱组比较,联合组lncRNA HOTAIR表达量降低(P<0.05)。与转染组比较,联合组lncRNA HOTAIR表达量升高(P<0.05)。与空白组比较,次乌头原碱组细胞凋亡率及Beclin1、P62、LC3 Ⅰ蛋白相对表达量升高,GFP-LC3斑点数增加(均P<0.05); 转染组细胞凋亡率及Beclin1、P62、LC3 Ⅰ蛋白表达量降低,GFP-LC3斑点数减少(均P<0.05)。与次乌头原碱组比较,联合组细胞凋亡率及Beclin1、P62、LC3 Ⅰ蛋白相对表达量降低,GFP-LC3斑点数减少(均P<0.05)。与转染组比较,联合组细胞凋亡率及Beclin1、P62、LC3 Ⅰ蛋白相对表达量升高,GFP-LC3斑点数增加(均P<0.05)。结论:次乌头原碱可能通过诱导人肝癌SK-Hep-1细胞过度自噬而促进细胞凋亡,其作用机制可能与上调LncRNA HOTAIR表达有关。
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 concentration 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 increased,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 Beclin1,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 Ⅰ protein 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.

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备注/Memo

备注/Memo:
基金项目:江苏省中医药科技发展计划项目(YB2020070); 江苏省南通市中医医疗联盟科研课题(TZYK202218)
更新日期/Last Update: 2024-06-05