Department of Pathology, Affiliated Hospital of Nantong University, No. 20 Xisi Road, Nantong, 226001, Jiangsu, People's Republic of China.
Department of Chemotherapy, Affiliated Hospital of Nantong University, Nantong, Jiangsu, People's Republic of China.
Mol Cell Biochem. 2018 Aug;445(1-2):123-134. doi: 10.1007/s11010-017-3258-8. Epub 2018 Jan 11.
Phosphofructokinase-2/fructose-2, 6-bisphosphatase 3 (PFKFB3) catalyzes the synthesis of F2,6BP, which is an allosteric activator of 6-phosphofructo-1-kinase (PFK-1): the rate-limiting enzyme of glycolysis. During tumorigenesis, PFKFB3 increases glycolysis, angiogenesis, and tumor progression. In this study, our aim was to investigate the significance of PFKFB3 and Ki67 in human lung adenocarcinomas and to target PFKFB3 as a therapeutic strategy. In this study, we determined the expression levels of PFKFB3 mRNA and proteins in cancerous and normal lung adenocarcinomas by quantitative reverse transcription PCR (qRT-PCR), Western blot analysis, and tissue microarray immunohistochemistry analysis, respectively. In human adenocarcinoma tissues, PFKFB3 and Ki67 protein levels were related to the clinical characteristics and overall survival. Both PFKFB3 mRNA and protein were significantly higher in lung adenocarcinoma cells (all P < 0.05). A high expression of PFKFB3 and Ki67 were associated with the degree of differentiation, TNM staging, lymph node metastasis, and survival. A high expression of PFKFB3 protein was an independent prognostic marker in lung adenocarcinoma. Subsequently, 1-(4-pyridinyl)-3-(2-quinolinyl)-2-propen-1-one (PFK15) was used as a selective antagonist of PFKFB3. Glycolytic flux was determined by measuring glucose uptake, F2,6BP, and lactate production. Cell viability, cell cycle, cell apoptosis, cell migration, and invasion were analyzed by MTT, flow cytometry, Western blot analysis, wound healing assay, and transwell chamber assay. By targeting PFKFB3, it inhibited cell viability and glycolytic activity. It also caused apoptosis and induced cell cycle arrest. Furthermore, the migration and invasion of A549 cells was inhibited. We conclude that PFKFB3 bears an oncogene-like regulatory element in lung adenocarcinoma progression. In the treatment of lung adenocarcinoma, targeting PFKFB3 would be a promising therapeutic strategy.
磷酸果糖激酶-2/果糖-2,6-二磷酸酶 3（PFKFB3）催化 F2,6BP 的合成，F2,6BP 是 6-磷酸果糖-1-激酶（PFK-1）的别构激活剂：糖酵解的限速酶。在肿瘤发生过程中，PFKFB3 增加糖酵解、血管生成和肿瘤进展。在这项研究中，我们的目的是研究 PFKFB3 和 Ki67 在人类肺腺癌中的意义，并将 PFKFB3 作为一种治疗策略。在这项研究中，我们通过定量逆转录 PCR（qRT-PCR）、Western blot 分析和组织微阵列免疫组织化学分析，分别确定了 PFKFB3 mRNA 和蛋白质在癌性和正常肺腺癌中的表达水平。在人类腺癌组织中，PFKFB3 和 Ki67 蛋白水平与临床特征和总生存期相关。PFKFB3 和 Ki67 在肺腺癌细胞中的表达均显著升高（均 P<0.05）。PFKFB3 蛋白高表达与分化程度、TNM 分期、淋巴结转移和生存有关。PFKFB3 蛋白高表达是肺腺癌的独立预后标志物。随后，使用 1-(4-吡啶基)-3-(2-喹啉基)-2-丙烯-1-酮（PFK15）作为 PFKFB3 的选择性拮抗剂。通过测量葡萄糖摄取、F2,6BP 和乳酸生成来确定糖酵解通量。通过 MTT、流式细胞术、Western blot 分析、划痕愈合试验和 Transwell 室试验分析细胞活力、细胞周期、细胞凋亡、细胞迁移和侵袭。通过靶向 PFKFB3，抑制细胞活力和糖酵解活性。它还导致细胞凋亡并诱导细胞周期停滞。此外，A549 细胞的迁移和侵袭受到抑制。我们得出结论，PFKFB3 在肺腺癌进展中具有癌基因样调节元件。在肺腺癌的治疗中，靶向 PFKFB3 将是一种有前途的治疗策略。
