Department of Medical Oncology, Nanyang First People's Hospital, Nanyang 473012, China.
Department of Proctology, Nanyang First People's Hospital, Nanyang 473012, China.
Life Sci. 2019 Sep 1;232:116679. doi: 10.1016/j.lfs.2019.116679. Epub 2019 Jul 21.
Amplified in liver cancer 1 gene (ALC1), a recently identified oncogene, was reported to be overexpressed in esophageal cancer cell lines and identified as a target oncogene in esophageal cancer pathogenesis. However, little literature is available to illustrate its significance in cisplatin resistance of esophageal squamous cell carcinoma (ESCC) cells. The aim of the current study was to investigate the effect of ALC1 on cisplatin cytotoxicity of ESCC cells and to study the potential mechanisms.
ALC1 at mRNA and protein levels were detected by qRT-PCR and western blot, respectively. Cell viability was evaluated using CCK-8 assay. Apoptosis was assessed using caspase-3/7 activity assay and flow cytometry analysis. Glycolysis level was evaluated by measuring glucose consumption and lactate production. The protein levels of p-protein kinase B (Akt) and Akt were determined by western blot.
ALC1 was highly expressed in ESCC cells compared with human normal esophageal epithelial Het-1A cells. ALC1 knockdown suppressed the viability, induced apoptosis and enhanced cisplatin cytotoxicity in ESCC cells. In addition, ALC1 knockdown inhibited glycolysis and inactivated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway in ESCC cells. Mechanistically, activation of the PI3K/Akt pathway by 740Y-P blocked the effects of ALC1 knockdown on cisplatin cytotoxicity and glycolysis in ESCC cells. In contrast, inhibition of the PI3K/Akt pathway by LY294002 or glycolysis by 2-deoxyglucose resisted the effect of ALC1 overexpression on cisplatin cytotoxicity in ESCC cells.
ALC1 knockdown enhanced cisplatin cytotoxicity of ESCC cells by inhibition of glycolysis through inactivation of the PI3K/Akt pathway.
最近发现的癌基因扩增在肝癌 1 基因（ALC1）中被报道在食管癌细胞系中过度表达，并被确定为食管癌发病机制中的靶癌基因。然而，很少有文献说明它在食管鳞状细胞癌（ESCC）细胞顺铂耐药中的意义。本研究旨在探讨 ALC1 对 ESCC 细胞顺铂细胞毒性的影响，并研究其潜在机制。
通过 qRT-PCR 和 Western blot 分别检测 ALC1 在 mRNA 和蛋白水平的表达。用 CCK-8 法评估细胞活力。用 caspase-3/7 活性测定和流式细胞术分析评估细胞凋亡。通过测量葡萄糖消耗和乳酸产生来评估糖酵解水平。用 Western blot 法测定磷酸化蛋白激酶 B（Akt）和 Akt 的蛋白水平。
与正常人食管上皮细胞 Het-1A 相比，ALC1 在 ESCC 细胞中高表达。ALC1 敲低抑制 ESCC 细胞的活力，诱导细胞凋亡，并增强顺铂的细胞毒性。此外，ALC1 敲低抑制 ESCC 细胞的糖酵解并使磷脂酰肌醇 3-激酶（PI3K）/Akt 通路失活。在机制上，740Y-P 激活 PI3K/Akt 通路阻断了 ALC1 敲低对 ESCC 细胞顺铂细胞毒性和糖酵解的影响。相反，LY294002 抑制 PI3K/Akt 通路或 2-脱氧葡萄糖抑制糖酵解，抵抗 ALC1 过表达对 ESCC 细胞顺铂细胞毒性的影响。
ALC1 敲低通过抑制 PI3K/Akt 通路失活抑制糖酵解，增强 ESCC 细胞顺铂的细胞毒性。
