Department of General Surgery, The Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi 710004 China.
Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai 200032 China.
Int J Biol Sci. 2018 Jan 11;14(1):100-110. doi: 10.7150/ijbs.22555. eCollection 2018.
In this study, we investigated whether the metabolic alteration of (CAFs) occurs via miR-21 remodeling and the effect of this alteration on pancreatic cancer cells. CAFs and normal fibroblasts (NFs) were isolated and cultured. Glucose consumption and lactic acid production were tested, and lactate dehydrogenase (LDHA), pyruvate kinase m2 (PKM2), and miR-21 expression were examined. The level of glycolysis in CAFs was determined after treatment with a miR-21 inhibitor. Primary miR-21-NC CAFs and miR-21-inhibitor CAFs were indirectly co-cultured with BxPc-3 , and the invasion capacity of these cells was determined. The aerobic oxidation index of cancer cells and the expression of succinodehydrogenase (SDH) and fumarate hydratase (FH) were assessed. Compared with NFs, CAFs showed enhanced glucose uptake capacity, lactic acid production, and elevated LDHA, PKM2, and miR-21 expression. After miR-21 inhibitor treatment, the extent of glycolysis in CAFs was reduced. After indirect co-culture with CAFs, oxidative phosphorylation and SDH, FH, and MCT expression increased in BxPc-3 cells. After co-culture with miR-21-inhibitor-CAFs, oxidative phosphorylation and invasion ability of the pancreatic cancer cells decreased. MiR-21 was involved in metabolic alteration of CAFs and affected the development of cancer cells. This metabolic alteration may be an important mechanism by which the microenvironment promotes tumor progression in a nonvascular manner.
在这项研究中，我们研究了代谢改变是否通过 miR-21 重塑发生，以及这种改变对胰腺癌细胞的影响。分离并培养了成纤维细胞（CAFs）和正常成纤维细胞（NFs）。检测了葡萄糖消耗和乳酸生成，并检查了乳酸脱氢酶（LDHA）、丙酮酸激酶 m2（PKM2）和 miR-21 的表达。用 miR-21 抑制剂处理 CAFs 后，测定了糖酵解的水平。用原发性 miR-21-NC CAFs 和 miR-21 抑制剂 CAFs 间接共培养 BxPc-3，并测定这些细胞的侵袭能力。评估了癌细胞的需氧氧化指数以及琥珀酸脱氢酶（SDH）和延胡索酸水合酶（FH）的表达。与 NFs 相比，CAFs 表现出增强的葡萄糖摄取能力、乳酸生成以及升高的 LDHA、PKM2 和 miR-21 表达。用 miR-21 抑制剂处理后，CAFs 中的糖酵解程度降低。与 CAFs 间接共培养后，BxPc-3 细胞中的氧化磷酸化以及 SDH、FH 和 MCT 的表达增加。与 miR-21 抑制剂-CAFs 共培养后，氧化磷酸化和胰腺癌细胞的侵袭能力降低。miR-21 参与了 CAFs 的代谢改变，并影响了癌细胞的发育。这种代谢改变可能是微环境以非血管方式促进肿瘤进展的重要机制。
