Kase Satoru, Yoshida Kazuhiko, Nakayama Keiichi I, Nakayama Keiko, Ikeda Hiromi, Harada Takayuki, Harada Chikako, Ohgami Kazuhiro, Shiratori Kenji, Ohno Shigeaki
Department of Ophthalmology and Visual Sciences, Hokkaido University Graduate School of Medicine, N15 W7, Kita-ku, Sapporo 060-8638, Japan.
Int J Mol Med. 2005 Aug;16(2):257-62.
Cellular distribution of the p27(KIP1) protein and its phosphorylation on threonine (T) 187 in mouse retinas from three stages of development, and retinoblastoma were examined. Retinas in C57Bl6 mice at embryonic day (E) 14, postnatal day (P) 1 and P11 were analyzed using immunohistochemistry with anti-p27(KIP1), threonine-187-phosphorylated p27(KIP1) (T187-phospho-p27), bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA) antibodies, and phosphorylated histon H3 (pHiston H3), which is a marker for cells in M phase. p27(KIP1) knockout (-/-) mice and human retinoblastoma were also analyzed. T187-phospho-p27 was detected in the outermost layer of the retina, whereas several neuroblastic cells expressed p27(KIP1) at E14. Many neuroblastic cells expressed BrdU in the middle layer. At P1, p27(KIP1) was detected in the ganglion cell layer and neuroblastic layer. T187-phospho-p27 was detected in the outermost layer, and that was localized in mitotic cells that also showed pHiston H3-positive. At P11, p27(KIP1) was detected in the inner nuclear layer, whereas T187-phospho-p27-positive or mitotic cells were not. BrdU positive nuclei were not detected in wild-type but were noted in the inner nuclear layer and the outer nuclear layer of the p27(KIP1) -/- mice retina at P11. In retinoblastoma, tumor cells formed numerous rosettes with Flexner-Wintersteiner rosettes. Several pHiston H3 -positive nuclei were noted in the tumor cells forming Flexner-Wintersteiner rosettes. Several T187-phospho-p27-positive nuclei were also detected in the mitotic cells forming Flexner-Wintersteiner rosettes. PCNA was expressed in rosette-forming cells. In conclusion, T187-phospho-p27(KIP1) was correlated with M phase of the cell cycle in the developing retina and retinoblastoma.
研究了p27（KIP1）蛋白在小鼠视网膜发育三个阶段以及视网膜母细胞瘤中的细胞分布及其苏氨酸（T）187位点的磷酸化情况。使用抗p27（KIP1）、苏氨酸-187磷酸化的p27（KIP1）（T187-磷酸化-p27）、溴脱氧尿苷（BrdU）、增殖细胞核抗原（PCNA）抗体以及有丝分裂期细胞标志物磷酸化组蛋白H3（p组蛋白H3）的免疫组织化学方法，分析了C57Bl6小鼠胚胎期第14天（E14）、出生后第1天（P1）和第11天（P11）的视网膜。还分析了p27（KIP1）基因敲除（-/-）小鼠和人视网膜母细胞瘤。在视网膜最外层检测到T187-磷酸化-p27，而在E14时几个成神经细胞表达p27（KIP1）。许多成神经细胞在中层表达BrdU。在P1时，在神经节细胞层和成神经细胞层检测到p27（KIP1）。在最外层检测到T187-磷酸化-p27，其定位于也呈p组蛋白H3阳性的有丝分裂细胞中。在P11时，在内核层检测到p27（KIP1），而未检测到T187-磷酸化-p27阳性或有丝分裂细胞。在野生型中未检测到BrdU阳性细胞核，但在P11时在p27（KIP1）-/-小鼠视网膜的内核层和外核层中观察到BrdU阳性细胞核。在视网膜母细胞瘤中，肿瘤细胞形成许多具有Flexner-Wintersteiner菊形团的玫瑰花结。在形成Flexner-Wintersteiner菊形团的肿瘤细胞中观察到几个p组蛋白H3阳性细胞核。在形成Flexner-Wintersteiner菊形团的有丝分裂细胞中也检测到几个T187-磷酸化-p27阳性细胞核。PCNA在形成玫瑰花结的细胞中表达。总之，T187-磷酸化-p27（KIP1）与发育中的视网膜和视网膜母细胞瘤的细胞周期M期相关。
