Si-HOTAIR对肝癌裸鼠移植瘤凋亡相关因子Bcl-2和caspase-3表达的影响

Effect of si-HOTAIR on the expression of apoptosis-related factors Bcl-2 and caspase-3 in hepatoma nude mice with xenograft tumors

【目的】探讨长链非编码HOTAIR对肝癌细胞凋亡相关因子Bcl-2和caspase-3表达的影响。【方法】18只裸鼠随机分为3组:对照组、空载体组和si-HOTAIR组,分组6只,分别接种人肝癌HepG2细胞株、转染空白序列的人肝癌HepG2细胞株和转染si-HOTAIR的人肝癌HepG2细胞株;采用流式细胞术检测瘤体内肝癌细胞的凋亡百分率;定量即时聚合酶链反应、蛋白质印迹法及免疫组织化学分别检测瘤体组织中Bcl-2、caspase-3 mRNA和蛋白的表达情况。【结果】Si-HOTAIR组肝癌细胞凋亡率较对照组、空载体组明显升高(P<0.05);与对照组和空载体组比较,si-HOTAIR组瘤组织内Bcl-2 mRNA和蛋白表达下调明显,则caspase-3 mRNA和蛋白表达显著上调(P<0.05)。【结论】沉默HOTAIR基因后可通过下调Bcl-2和上调caspase-3的表达促进肝癌细胞凋亡。

【Objective】To explore the effect of si-HOTAIR on the expression of apoptosis-related factors Bcl-2 and caspase-3 in hepatoma nude mice with xenograft tumors.【Methods】Eighteen nude mice were randomly divided into three groups:the control group,the empty carrier group and si-HOTAIR group,which were respectively inoculated with HepG2 liver cancer cells,HepG2 cell line transfected with blank sequence and HepG2 cell line transfected with si-HOTAIR.Flow Cytometry was used to detect the percentage of apoptosis of hepatocellular carcinoma cells in hepatocellular carcinoma tissues,and qRt-PCR,Western blot and immunohistochemistry were used to respectively detect the expression of Bcl-2,caspase-3 mRNA and protein in hepatocellular carcinoma tissues.【Results】The percentage of apoptosis of hepatoma cells in the si-hOTAIR group was significantly higher than those in the control group and the empty carrier group(P<0.05).Compared with the control group and the empty carrier group,the expression of Bcl-2 mRNA and protein in the tumor tissue of si-HOTAIR group was significantly decreased,while the expression of caspase-3 mRNA and protein was significantly increased(P<0.05).【Conclusion】The apoptosis of hepatoma cells can be promoted by the low expression of Bcl-2 and high expression of caspase-3 that can be regulated by silencing HOTAIR gene.