【摘要】 目的：研究长链脂酰辅酶A合成酶3(ACSL3)在代谢应激下对肝细胞癌（HCC）细胞Huh7存活的影响。方法：从TCGALICH数据库下载HCC患者RNA表达谱和临床数据，统计ACSL3在HCC患者中的表达情况，绘制Kaplan-Meier生存曲线；在广西医科大学第一附属医院的HCC队列中收集44例原发性HCC患者组织和癌旁组织样本，通过免疫组化检测ACSL3的表达；利用siRNA和CRISPR/CAS 9技术沉默/敲除Huh7细胞中的ACSL3基因，通过细胞计数试剂盒（CCK-8）实验、克隆形成实验和Transwell侵袭实验检测ACSL3敲除对Huh7细胞增殖和侵袭能力的影响。在完全培养基和葡萄糖缺乏条件下，通过细胞流式术检测对照组细胞和ACSL3沉默/敲除组细胞的死亡，并检测细胞中ATP、β-羟基丁酸的变化。结果：与正常肝组织相比，HCC组织的ACSL3表达较高（P<0.05），且ACSL3高表达患者生存期较短（P<0.05）。敲除ACSL3不影响Huh7细胞的增殖能力、克隆形成能力以及侵袭能力（P>0.05）。在葡萄糖缺乏下，ACSL3沉默/敲除导致Huh7细胞的存活率显著下降（P<0.05）,ATP产生水平下降以及β-羟基丁酸生成受抑制（P<0.05）。结论：通过抑制β氧化，敲除ACSL3促进了HCC细胞死于葡萄糖缺乏。更多还原

【Abstract】 Objective: To investigate the effect of long-chain acyl-CoA synthetase 3(ACSL3) on the survival of Huh7 cells of hepatocellular carcinoma(HCC) under metabolic stress. Methods: The RNA expression profile and clinical data of HCC patients were downloaded from the TCGA-LICH database to analyze the expression of ACSL3 in HCC patients and draw Kaplan-Meier survival curve. From the liver cancer cohort of the First Affiliated Hospital of Guangxi Medical University, 44 primary liver cancer tissues and paracancerous tissues in HCC patients were collected, and the expression of ACSL3 was detected by immunohistochemistry. ACSL3 expression in Huh7 cells was silenced/knocked-out by siRNA and CRISPR/CAS9 technology; the effect of ACSL3 knockout on the proliferation and invasion of Huh7 cells were detected by cell counting kit(CCK-8) assay, clonal formation assay and Transwell invasion assay. Under full medium and glucose deficiency culture conditions, cell death in control group and ACSL3 silencing or knockout group was measured by flow cytometry; and level changes of ATP and β-hydroxybutyrate in cells were detected. Results: Compared with normal liver tissues, the expression of ACSL3 was higher in HCC tissues(P<0.05); and patients with high ACSL3 exhibited shorter survival time(P<0.05). ACSL3 knockout did not affect cell proliferation, clonogenicity and invasion of Huh7 cells(P>0.05).Under the condition of glucose deficiency, ACSL3 silencing or knockout significantly decreased the survival rate of Huh7 cells(P<0.05), decreased ATP production and inhibited β-hydroxybutyrate production(P<0.05). Conclusion: Knockout of ACSL3 promotes the HCC cell death from glucose deficiency by inhibiting β-oxidation.更多还原