Lee Yuumi, Ito Yuko, Taniguchi Kohei, Nuri Takashi, Lee SangWoong, Ueda Koichi
From the Department of Plastic and Reconstructive Surgery, Osaka Medical and Pharmaceutical University, Osaka, Japan.
Department of General and Gastroenterological Surgery, Osaka Medical and Pharmaceutical University, Osaka, Japan.
Plast Reconstr Surg Glob Open. 2023 Aug 16;11(8):e5202. doi: 10.1097/GOX.0000000000005202. eCollection 2023 Aug.
A keloid is composed of several nodules, which are divided into two zones: the central zone (CZ; a hypoxic region) and the marginal zone (MZ; a normoxic region). Keloid nodules play a key role in energy metabolic activity for continuous growth by increasing in number and total area. In this study, we aimed to investigate the roles of the zones in the execution of the Warburg effect and identify which microRNAs regulate this phenomenon in keloid tissue.
Eleven keloids from patients were used. Using immunohistochemical analysis, 179 nodules were randomly chosen from these keloids to identify glycolytic enzymes, autophagic markers, pyruvate kinase M (PKM) 1/2, and polypyrimidine tract binding protein 1 (PTBP1). Western blot and qRT-PCR tests were also performed for PKM, PTBP1, and microRNAs (miR-133b and miR-200b, c).
Immunohistochemical analysis showed that the expression of the autophagic (LC3, p62) and glycolytic (GLUT1, HK2) were significantly higher in the CZ than in the MZ. PKM2 expression was significantly higher than PKM1 expression in keloid nodules. Furthermore, PKM2 expression was higher in the CZ than in the MZ. However, PKM1 and PTBP1 expression levels were higher in the MZ than in the CZ. The qRT-PCR analysis showed that miR-133b-3p was moderately downregulated in the keloids compared with its expression in the normal skin tissue.
The Warburg effect occurred individually in nodules. The MZ presented PKM2-positive fibroblasts produced by activated PTBP1. In the CZ, PKM2-positive fibroblasts produced lactate. MiR-133b-3p was predicted to control the Warburg effect in keloids.
瘢痕疙瘩由多个结节组成，这些结节分为两个区域：中央区（CZ；缺氧区域）和边缘区（MZ；常氧区域）。瘢痕疙瘩结节通过数量和总面积的增加，在持续生长的能量代谢活动中起关键作用。在本研究中，我们旨在探讨这些区域在执行瓦伯格效应中的作用，并确定哪些微小RNA调节瘢痕疙瘩组织中的这一现象。
使用来自患者的11个瘢痕疙瘩。通过免疫组织化学分析，从这些瘢痕疙瘩中随机选取179个结节，以鉴定糖酵解酶、自噬标志物、丙酮酸激酶M（PKM）1/2和多嘧啶序列结合蛋白1（PTBP1）。还对PKM、PTBP1和微小RNA（miR-133b和miR-200b、c）进行了蛋白质印迹和qRT-PCR检测。
免疫组织化学分析显示，自噬（LC3、p62）和糖酵解（GLUT1、HK2）标志物在中央区的表达明显高于边缘区。瘢痕疙瘩结节中PKM2的表达明显高于PKM1的表达。此外，PKM2在中央区的表达高于边缘区。然而，PKM1和PTBP1的表达水平在边缘区高于中央区。qRT-PCR分析显示，与正常皮肤组织中的表达相比，miR-133b-3p在瘢痕疙瘩中中度下调。
瓦伯格效应在结节中单独发生。边缘区呈现由活化的PTBP1产生的PKM2阳性成纤维细胞。在中央区，PKM2阳性成纤维细胞产生乳酸。预测miR-133b-3p可控制瘢痕疙瘩中的瓦伯格效应。
