Zhou Y, Lee H S, Kooshesh F, Fujisawa H, Sauder D N, Kondo S
Division of Dermatology, Sunnybrook Health Science Centre, University of Toronto, Canada.
Exp Dermatol. 1996 Jun;5(3):138-44. doi: 10.1111/j.1600-0625.1996.tb00108.x.
Keratinocyte growth factor (KGF) and its receptor (KGFR) are thought to play important roles in normal keratinocyte growth and differentiation. Since UVB radiation is known to influence keratinocyte growth, we sought to determine whether UVB would alter the expression of KGF and KGFR. Using a reverse-transcription coupled polymerase chain reaction (RT-PCR), the present study examined the expression of KGF and KGFR mRNA in cultured normal human keratinocytes exposed to UVB irradiation. Total cellular RNA was extracted from cultured keratinocytes at various time points after irradiation, reverse transcribed and used for PCR amplification using primers specific for KGF and KGFR. Constitutive expression of KGFR mRNA, but not KGF mRNA, was detected in normal cultured human keratinocytes. After UVB irradiation at 300 J/m2, the KGF mRNA remained undetectable while the KGFR mRNA level was significantly decreased. The down-regulation of KGFR mRNA expression was also confirmed by Northern blot analysis. Immunohistochemical studies demonstrated a decreased positive signal of KGFR in human keratinocytes after UVB irradiation. Our results suggest a possible role for the KGF-KGFR signalling pathway in the skin after exposure to UVB, and that UVB-induced growth inhibition of keratinocytes in hyperproliferative skin disorders may be related to downregulation of KGFR.
角质形成细胞生长因子(KGF)及其受体(KGFR)被认为在正常角质形成细胞的生长和分化中发挥重要作用。由于已知紫外线B(UVB)辐射会影响角质形成细胞的生长,我们试图确定UVB是否会改变KGF和KGFR的表达。本研究使用逆转录聚合酶链反应(RT-PCR),检测了暴露于UVB照射的培养正常人角质形成细胞中KGF和KGFR mRNA的表达。在照射后的不同时间点,从培养的角质形成细胞中提取总细胞RNA,逆转录后使用针对KGF和KGFR的特异性引物进行PCR扩增。在正常培养的人角质形成细胞中检测到KGFR mRNA的组成性表达,但未检测到KGF mRNA。在300 J/m2的UVB照射后,KGF mRNA仍未检测到,而KGFR mRNA水平显著降低。Northern印迹分析也证实了KGFR mRNA表达的下调。免疫组织化学研究表明,UVB照射后人角质形成细胞中KGFR的阳性信号降低。我们的结果表明,KGF-KGFR信号通路在暴露于UVB后的皮肤中可能发挥作用,并且UVB诱导的增生性皮肤病中角质形成细胞的生长抑制可能与KGFR的下调有关。