State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China; Department of Cariology and Endodontics II, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Am J Pathol. 2021 May;191(5):857-871. doi: 10.1016/j.ajpath.2021.02.010. Epub 2021 Feb 26.
To investigate the role of glycolysis and the M2 isoform of pyruvate kinase (PKM2) in odontogenic keratocysts (OKCs), the glycolytic flux of primary odontogenic keratocyst fibroblasts (OKC-Fs) and normal oral mucosa fibroblasts (OM-Fs) was determined by glucose uptake, lactate production, and cell proliferation assays. Wound healing assay and Matrigel-coated chamber system were used to investigate the effects of PKM2 on migration and invasion capacities of OKC-Fs. Co-culture of OKC-Fs with osteoclast precursors (RAW264.7 cells) was used to clarify the role of glycolysis in the osteoclastogenic effects of OKC-Fs. In addition, hypoxia-inducible factor 1α and some key enzymes related to glycolysis, including PKM2, 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3, hexokinase 2, and lactate dehydrogenase A, were detected to assess the activation of glycolysis in OKC stroma by immunohistochemistry. Results showed that the glucose uptake and lactate production were significantly higher in OKC-Fs than OM-Fs. PKM2 was elevated in OKC-Fs compared with that in OM-Fs. PKM2 significantly regulated glycolysis, proliferation, migration, invasion, and osteoclastogenic effects of OKC-Fs. Additionally hypoxia-inducible factor 1α, 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3, hexokinase 2, and lactate dehydrogenase A were markedly overexpressed in OKC stroma, and correlated with PKM2. Moreover, the expression of PKM2 was regulated by oxygen concentration in vitro. In sum, PKM2-mediated glycolysis regulated the growth, aggressiveness, and osteoclastogenesis of OKC.
为了研究糖酵解和丙酮酸激酶（PKM2）M2 同工型在牙源性角化囊肿（OKC）中的作用，通过葡萄糖摄取、乳酸生成和细胞增殖测定来确定原代牙源性角化囊肿成纤维细胞（OKC-Fs）和正常口腔黏膜成纤维细胞（OM-Fs）的糖酵解通量。通过划痕愈合实验和 Matrigel 包被室系统来研究 PKM2 对 OKC-Fs 迁移和侵袭能力的影响。将 OKC-Fs 与破骨细胞前体（RAW264.7 细胞）共培养，以阐明糖酵解在 OKC-Fs 破骨细胞生成作用中的作用。此外，通过免疫组织化学检测缺氧诱导因子 1α 和一些与糖酵解相关的关键酶，包括 PKM2、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3、己糖激酶 2 和乳酸脱氢酶 A，以评估 OKC 基质中糖酵解的激活情况。结果表明，OKC-Fs 的葡萄糖摄取和乳酸生成明显高于 OM-Fs。与 OM-Fs 相比，OKC-Fs 中的 PKM2 升高。PKM2 显著调节 OKC-Fs 的糖酵解、增殖、迁移、侵袭和破骨细胞生成作用。此外，缺氧诱导因子 1α、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3、己糖激酶 2 和乳酸脱氢酶 A 在 OKC 基质中明显过表达，并与 PKM2 相关。此外，PKM2 的表达在体外受氧浓度调节。总之，PKM2 介导的糖酵解调节 OKC 的生长、侵袭性和破骨细胞生成。
