Department of Respiratory Medicine, Nanjing First Hospital, Nanjing Medical University, No. 68 Changle Road, Qinhuai District, Nanjing, 210001, People's Republic of China.
Lipids Health Dis. 2019 May 24;18(1):118. doi: 10.1186/s12944-019-1058-8.
Fatty acid synthase (FASN) is overexpressed in most human carcinomas, including non-small cell lung cancer (NSCLC), and contributes to poor prognosis. An increasing number of studies have highlighted the potential function of FASN as both a biomarker and therapeutic target for cancers. However, the underlying molecular mechanisms of FASN in glucose metabolism and the malignant biological behavior of NSCLC remain the subjects of intensive investigation.
FASN expression was depleted by FASN-siRNA in A549 and NCI-H1299 cell lines to detect the function of glucose metabolism and the malignant biological behavior of NSCLC cells. Western-blot and qPCR were applied to determine the expressions of FASN, t-AKT, p-AKT, t-ERK, p-ERK, PKM2, HK2 and AZGP1. ATP and lactate were detected to determine the activation of glucose metabolism. CCK8 and transwell assays were used to detect the proliferation, invasion, and migration capacity of the two types of NSCLC cells. The xenograft mouse model was used to evaluate tumor weights after suppression of FASN.
LV-FASN-siRNA and its control lentiviral vector were successfully transfected into the two types of NSCLC cells (A549 and NCI-H1299). LV-FASN siRNA significantly suppressed FASN expression in both NSCLC cell types, and expressions of p-AKT, p-ERK, PKM2, and AZGP1 were also significantly decreased. Notably, the levels of ATP and lactate were significantly decreased after transfection with LV-FASN siRNA. The proliferation of both NSCLC cell types was decreased after suppression of FASN. The invasion and migration capacity of A549, but not NCI-H1299, were inhibited following down-regulation of FASN. In vivo, inhibition of FASN caused a marked animal tumor weight loss.
FASN was involved in glucose metabolism via down-regulation of the AKT/ERK pathway and eventually altered the malignant phenotype in lung cancer cells.
脂肪酸合酶（FASN）在大多数人类癌肿中过度表达，包括非小细胞肺癌（NSCLC），并导致预后不良。越来越多的研究强调了 FASN 作为癌症的生物标志物和治疗靶点的潜在功能。然而，FASN 在葡萄糖代谢和 NSCLC 恶性生物学行为中的潜在分子机制仍在深入研究中。
通过 FASN-siRNA 在 A549 和 NCI-H1299 细胞系中敲低 FASN 的表达，以检测 NSCLC 细胞葡萄糖代谢和恶性生物学行为的功能。Western blot 和 qPCR 用于确定 FASN、t-AKT、p-AKT、t-ERK、p-ERK、PKM2、HK2 和 AZGP1 的表达。检测 ATP 和乳酸以确定葡萄糖代谢的激活情况。CCK8 和 Transwell 测定用于检测两种类型的 NSCLC 细胞的增殖、侵袭和迁移能力。抑制 FASN 后，使用异种移植小鼠模型评估肿瘤重量。
LV-FASN-siRNA 及其对照慢病毒载体成功转染至两种类型的 NSCLC 细胞（A549 和 NCI-H1299）。LV-FASN siRNA 显著抑制两种 NSCLC 细胞类型中的 FASN 表达，p-AKT、p-ERK、PKM2 和 AZGP1 的表达也显著降低。值得注意的是，转染 LV-FASN siRNA 后，ATP 和乳酸的水平显著降低。抑制 FASN 后，两种 NSCLC 细胞类型的增殖均降低。FASN 下调后，A549 的侵袭和迁移能力受到抑制，但 NCI-H1299 则不受影响。体内，抑制 FASN 导致动物肿瘤重量明显减轻。
FASN 通过下调 AKT/ERK 通路参与葡萄糖代谢，最终改变肺癌细胞的恶性表型。
