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[目的]探讨SCD1对SMMC-7721细胞脂质代谢的影响及可能机制。[方法]肝癌细胞系SMMC-7721细胞培养,分为4组(对照组、CAY-10566处理组、CAY-10566联合Compound C共同处理组和CAY-10566联合氯喹共同处理组),通过MTT法和流式细胞术分别检测细胞增殖和凋亡情况,透射电子显微镜观察细胞自噬体情况。胆固醇(CE)和甘油三酯(TAG)检测试剂盒分别检测上述4组SMMC-7721细胞的CE和TAG水平。[结果]抑制SCD1后SMMC-7721细胞自噬体明显增加,相应地减少了细胞活力,增加了细胞凋亡率;抑制SCD1基础上抑制AMPK或自噬后上述细胞的自噬体均明显减少,相应地增加了细胞活力,减少了细胞凋亡率。CE和TAG检测结果表明:抑制SCD1明显减少SMMC-7721细胞CE和TAG水平,与对照组相比,差异均有统计学意义(P<0.05);抑制SCD1基础上抑制AMPK或自噬后SMMC-7721细胞CE和TAG水平均明显增加,与SCD1抑制剂组比较,差异均有统计学意义(P<0.05)。[结论 ]SCD1可通过下调AMPK途径负性调节自噬,增加SMMC-7721细胞CE和TAG水平,抑制细胞凋亡,促进细胞生存,可能影响肝癌细胞的形成及进展。
[Objective] To investigate the effect of SCD1 on lipid metabolism in SMMC-7721 cells and its possible mechanism. [Methods] The hepatoma cell line SMMC-7721 cells were cultured and divided into 4 groups (control group, CAY-10566 treatment group, CAY-10566 combined Compound C treatment group and CAY-10566 combined chloroquine co-treatment group) Cell proliferation and apoptosis were detected by flow cytometry. The autophagosomes were observed by transmission electron microscopy. Cholesterol (CE) and triglyceride (TAG) detection kit were used to detect CE and TAG levels in the above four groups of SMMC-7721 cells respectively. [Results] The autophagosome of SMMC-7721 cells was significantly increased after SCD1 was inhibited, correspondingly the cell viability was decreased and the apoptosis rate was increased. The inhibition of autophosphorylation of AMPK or autophagy on the basis of SCD1 significantly decreased the number of autophagosomes, Correspondingly increase the cell viability and reduce the rate of apoptosis. The results of CE and TAG showed that the inhibition of SCD1 significantly reduced the levels of CE and TAG in SMMC-7721 cells compared with the control group (P <0.05), and inhibited the expression of SCD1 on the basis of inhibition of AMPK or autophagy after SMMC- Compared with SCD1 inhibitor group, the levels of CE and TAG in 7721 cells were significantly increased (P <0.05). [Conclusion] SCD1 can negatively regulate autophagy by down-regulating AMPK pathway, increasing CE and TAG levels, inhibiting apoptosis and promoting cell survival in SMMC-7721 cells, which may affect the formation and progression of hepatocellular carcinoma cells.