Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
Arch Oral Biol. 2017 Nov;83:340-347. doi: 10.1016/j.archoralbio.2017.08.014. Epub 2017 Sep 2.
The purpose of this study was to investigate a difference in glycogen metabolism (glycogen synthesis and glycolysis) between the iodine stained (normal non-keartinized) and the unstained (dysplasctic/malignant) oral epithelium.
Twenty-one frozen tissue samples of iodine-stained and unstained mucosal tissue were obtained from 21 OSCC patients. Serial frozen sections were cut and examined with the hematoxylin-eosin and periodic acid-Schiff methods and immunohistochemical (IHC) staining for Ki67, P53, molecules associated with glycogenesis (i.e., glycogen synthase (GS) and phospho-glycogen synthase (PGS)), and molecules associated with glycogenolysis (i.e., glycogen phosphorylase isoenzyme BB (GPBB) examine the glycogen metabolism in OSCC. Additionally, in vitro study, the expression levels of GS and GPBB in the cultured cells were analyzed by immunofluorescent staining, Western blot analysis, and the real-time quantitative polymerase chain reaction (PCR).
There was no significant difference in GS and PGS immunoactivity between iodine stained and unstained area. On the other hand, significantly greater GPBB immunoreactivity was observed in the basal and parabasal layers of iodine-unstained epithelium, where higher positivity for p53 and Ki67 was also showed. Additionally, western blot analysis, immunofluorescent staining, and real-time quantitative PCR revealed that the oral squamous cancer cells exhibited greater expression of GPBB than normal epithelial cells.
The results of this study showed that GPBB expression, which resulted in up-regulation of glycogenolysis, is enhanced in oral dysplastic/malignant epithelium compared with non-keartinized normal epithelium, in spite of the fact that glycogenesis continues in both of them. Premalignant and malignant epithelial cells consume greater quantities of energy due to their increased proliferation, and hence, exhaust their glycogen stores, which resulting in negative stain reaction with iodine solution.
本研究旨在探讨碘染色(正常非角化)和未染色(发育不良/恶性)口腔上皮组织中糖原代谢(糖原合成和糖酵解)的差异。
从 21 例口腔鳞状细胞癌(OSCC)患者中获得 21 例碘染色和未染色的冷冻组织样本。对连续冷冻切片进行苏木精-伊红和过碘酸-希夫(PAS)染色以及 Ki67、P53、与糖原生成相关的分子(即糖原合酶(GS)和磷酸化糖原合酶(PGS))和与糖原分解相关的分子(即糖原磷酸化酶同工酶 BB(GPBB))的免疫组化(IHC)染色,以检查 OSCC 中的糖原代谢。此外,在体外研究中,通过免疫荧光染色、Western blot 分析和实时定量聚合酶链反应(PCR)分析培养细胞中 GS 和 GPBB 的表达水平。
碘染色和未染色区域的 GS 和 PGS 免疫活性无显著差异。另一方面,在未染色的上皮基底层和副基底层中,GPBB 免疫反应性显著增强,同时 p53 和 Ki67 的阳性率也较高。此外,Western blot 分析、免疫荧光染色和实时定量 PCR 显示,口腔鳞状癌细胞的 GPBB 表达高于正常上皮细胞。
本研究结果表明,尽管糖原生成在两者中都继续进行,但 GPBB 的表达导致糖原分解的上调,在口腔发育不良/恶性上皮组织中增强,而不是在非角化正常上皮组织中增强。由于增殖增加,癌前和恶性上皮细胞消耗更多的能量,因此耗尽糖原储存,导致碘溶液的阴性染色反应。