Institute of Stomatology, Nanjing Medical University, Nanjing, China; Department of Periodontology, College of Stomatology, Nanjing Medical University, Nanjing, China.
J Periodontal Res. 2013 Dec;48(6):740-7. doi: 10.1111/jre.12064. Epub 2013 Mar 25.
Keratinocyte growth factor (KGF) and its receptor (KGFR) are involved in hyperplastic diseases. This study explored the effect of intercellular communication on KGF and KGFR in cocultured/monocultured gingival fibroblasts and keratinocytes following treatment with nifedipine.
Human gingival fibroblasts and keratinocytes were monocultured and cocultured, respectively. MTT was used to investigate the effects of nifedipine on the proliferation of gingival fibroblasts and keratinocytes. Monoculture and coculture systems were treated with different concentrations (0, 0.2 or 20 μg/mL) of nifedipine, and the expression of KGF and KGFR mRNAs was examined by RT-PCR, whilst the secretion of KGF and the expression of KGFR on the membrane were analyzed using ELISA and flow cytometry, respectively.
Nifedipine (0, 0.2 and 20 μg/mL) had no influence on cell proliferation within 3 d. KGF and KGFR mRNAs were up-regulated, but only in the cocultures. In coculture, the secretion of KGF was significantly increased by nifedipine, while it was only significantly up-regulated by 20 μg/mL of nifedipine in monoculture. Moreover, the level of KGFR protein in the membrane was significantly increased by 20 μg/mL of nifedipine in monocultures, while it was significantly down-regulated by 20 μg/mL of nifedipine in cocultures.
The expression of KGF and KGFR are influenced by the interplay of gingival keratinocytes and fibroblasts. Epithelial keratinocytes and mesenchymal fibroblasts may interplay to dynamically regulate gene expression, which may have an effect on the gingival condition following treatment with nifedipine.
角质细胞生长因子（KGF）及其受体（KGFR）参与了增生性疾病。本研究旨在探讨细胞间通讯对硝苯地平处理后共培养/单培养牙龈成纤维细胞和角质细胞中 KGF 和 KGFR 的影响。
分别对人牙龈成纤维细胞和角质细胞进行单培养和共培养。采用 MTT 法检测硝苯地平对牙龈成纤维细胞和角质细胞增殖的影响。采用不同浓度（0、0.2 或 20μg/mL）的硝苯地平处理单培养和共培养系统，通过 RT-PCR 检测 KGF 和 KGFR mRNA 的表达，采用 ELISA 和流式细胞术分别分析 KGF 的分泌和膜上 KGFR 的表达。
硝苯地平（0、0.2 和 20μg/mL）在 3d 内对细胞增殖没有影响。KGF 和 KGFR mRNA 均上调，但仅在共培养中上调。在共培养中，硝苯地平显著增加了 KGF 的分泌，而在单培养中仅 20μg/mL 的硝苯地平显著增加了 KGF 的分泌。此外，20μg/mL 的硝苯地平显著增加了单培养中膜上 KGFR 蛋白的水平，而在共培养中则显著降低了 20μg/mL 的硝苯地平。
KGF 和 KGFR 的表达受牙龈角质形成细胞和成纤维细胞相互作用的影响。上皮角质形成细胞和间充质成纤维细胞可能相互作用以动态调节基因表达，这可能对硝苯地平治疗后的牙龈状况产生影响。
