摘要
目的探讨长链非编码RNA肝癌高表达转录本(highly up-regulated in liver cancer,LncRNA-HULC)通过稳定沉默信号调节因子1(silent information regulator 1,SIRT1)蛋白调控肝癌Hep G2细胞自噬和化疗耐药的机制。方法将实验Hep G2细胞分为3组:对照组(NOR组)、基因过表达组(O-HULC组)和基因抑制组(S-HULC组)。NOR组正常培养传代,O-HULC组转染HULC基因,S-HULC组将HULC基因抑制。应用免疫印迹法测定蛋白表达量,应用RT-QPCR法检测基因表达量。结果各组的肝癌Hep G2细胞自噬情况随药物添加时间延长而增加,凋亡率逐渐升高,与S-HULC组相比,O-HULC组和NOR组肝癌Hep G2细胞自噬情况显著增加,凋亡率降低,其中O-HULC组自噬情况高于NOR组,凋亡率低于NOR组(P<0.05)。与S-HULC组相比,NOR组和O-HULC组肝癌Hep G2细胞迁移数量和侵袭数量显著增加,其中O-HULC组显著高于NOR组(P<0.05)。与S-HULC组相比,NOR组和O-HULC组的泛素水解酶22(ubiquitin carboxyl-terminal hydrolase 22,USP22)、SIRT1、环氧化酶-2(cyclooxygenase-2,COX-2)及自噬相关蛋白微管相关蛋白1轻链3Ⅱ(Microtubule-associated protein1 light chain3,LC3-Ⅱ)蛋白表达量升高,其中O-HULC组均高于NOR组(P<0.05)。结论LncRNA-HULC可以稳定SIRT1调控细胞自噬降低细胞对化疗药物的敏感性,可能与信号通路“HULC-USP22-SIRT1/COX-2-LCⅡ”有关,此通路可能是开发肝癌化疗新增敏策略的新靶点。
Objective To investigate the mechanism of long non-coding RNA(LncRNA)highly up-regulated in liver cancer(HULC)regulating autophagy and chemoresistance of hepatoma Hep G2 cells by stabilizing silent information regulator1(SIRT1)protein.Methods Experimental Hep G2 cells were divided into three groups:control group(NOR group),gene overexpression group(O-HULC group)and gene suppression group(S-HULC group).The NOR group was normally cultured and subcultured,the O-HULC group was transfected with HULC gene,and the S-HULC group was suppressed with HULC gene.Western blot was used to determine the protein expression and RT-QPCR was used to detect gene expression.Results The autophagy of Hep G2 cells in each group increased with the extension of drug addition time,and the apoptosis rate gradually increased.Compared with the S-HULC group,the autophagy of Hep G2 cells in the O-HULC group and the NOR group significantly increased,and the apoptosis rate decreased,and the autophagy in the O-HULC group was higher than that in the NOR group.The apoptosis rate was lower than that of the NOR group(P<0.05).Compared with the S-HULC group,the migration number and invasion number of Hep G2 cells in the NOR group and the O-HULC group were significantly increased,and those in the O-HULC group were significantly higher than those in the NOR group(P<0.05).Compared with the S-HULC group,ubiquitin carboxyl-terminal hydrolase 22(USP22),SIRT1,Cyclooxygenase-2(COX-2)and microtubule-associated protein1 light chain 3Ⅱ(LC3-Ⅱ)protein expression levels were increased,and those in the O-HULC group were higher than those in the NOR group(P<0.05).Conclusion LncRNA-HULC can stabilize SIRT1,regulate autophagy and reduce the sensitivity of cells to chemotherapeutic drugs,which may be related to the signaling pathway HULC-USP22-SIRT1/COX-2-LCⅡ,and this pathway may be a new target for developing chemotherapeutic sensitization strategies for liver cancer.
作者
严冰冰
韩嘉晟
袁胜春
Yan Bingbing;Han Jiasheng;Yuan Shengchun
出处
《中国疗养医学》
2022年第2期129-132,共4页
Chinese Journal of Convalescent Medicine
关键词
自噬
蛋白
基因
化疗耐药
凋亡
迁移侵袭
Autophagy
Protein
Gene
Chemoresistance
Apoptosis
Migration and invasion
