干扰SULT2B1对Hepa1-6肝癌细胞凋亡的影响及机制研究

Effect of SULT2B1 Interference on Apoptosis of Mouse Hepa1-6 Hepatocellular Carcinoma Cells and Its Underlying Mechanism

目的研究羟基类固醇硫酸基转移酶SULT2B1对小鼠Hepa1-6肝癌细胞凋亡的影响及作用机制。方法SULT2B1慢病毒干扰载体感染Hepa1-6细胞(MOI=100),Real-time PCR检测其干扰效果。采用流式细胞仪通过Annexin V-APC/7-ADD染色检测干扰SULT2B1后Hepa1-6细胞凋亡率的改变,Western blot检测FAS和p-JNK的蛋白表达水平。结果构建的SULT2B1干扰载体可显著抑制Hepa1-6细胞中SULT2B1的表达(P<0.05)。Hepa1-6细胞中干扰SULT2B1后其凋亡率较对照组相比显著增加(P<0.05),干扰SULT2B1后可显著增加FAS和p-JNK的蛋白表达水平(P<0.05)。结论干扰SULT2B1可促进Hepa1-6细胞的凋亡,其作用主要与增加FAS和p-JNK的蛋白表达水平有关。

Objective To investigate the effect and possible mechanism of hydroxysteroid sulfotransferase 2B1( SULT2B1) on cell apoptosis of mouse hepatocellular carcinoma Hepa1-6 cells. Methods Hepa1-6 cells were infected with lentivirus- mediated SULT2B1 interference vectors at multiplicity of infection( MOI) of100. The mRNA levels of SULT2B1 in Hepa1- 6 cells treated with vectors were detected by real- time PCR analysis. The ratio of apoptotic cells were analyzed by flow cytometry after incubated with Annexin V- APC and 7- ADD. FAS and p- JNK protein levels were analyzed by western blot assay. Results SULT2B1 expression was inhibited significantly in Hepa1- 6 cells with constructed SULT2B1 interference vector( P <0. 05). The percentage of apoptosis was significantly increased in Hepa1- 6 with SULT2B1 interference,compared with control cells( P < 0. 05). Moreover,the protein levels of FAS and p- JNK were increased significantly in Hepa1- 6 cells with SULT2B1 knock- down( P < 0. 05). Conclusion Knock- down of SULT2B1 induced Hepa1- 6 cell apoptosis,which mechanism was mainly associated with up- regulation of FAS and p- JNK protein levels.