Expression of Inositol, 1,4,5-trisphosphate Receptor in the Nucleus of Human Benign and Malignant Squamous Cells and Its Significance

肌醇、1,4,5-三磷酸受体在人良恶性鳞状细胞核中的表达及其意义

• 批准号: 07671858
• 负责人: HARADA Takayuki
• 金额: $ 1.34万
• 依托单位: Shimane Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671858/
• 关键词: Inositol 1,4,5-trisphosphate receptor; squamous cell; cross-reactivity; nucleus; immunocytochemistry; human; 扁平上皮細胞; 免疫組織細胞化学; イノシトール三燐酸レセプター; 頭頚部癌細胞

Inositol 1,4,5-trisphosphate (IP_3) is one of the important messengers of the intracellular signal transduction, and controls the cytoplasmic flow of calcium ion in conjunction with its receptor (IP_3-R). The Ip_3-R protein and its coding gene had been revealed by Mikoshiba et al., and monoclonal antibodies reactive with mouse IP_3-R protein have been produced as well. Using cross-reactivity of these monoclonal antibodies (10A6,4C11,18A10) with the human counterpart we have shown the distribution of IP_3-R in human tissues and cells, especially in normal skin including benign squamous epithel, squamous cell carcinoma-tissues and cultured cell lines of squamous cell carcinoma. In the present researchproject we examined peculiar reactivity of 10A6 monoclonal antibody with an antigen in the nucleus. Presence of the antigen was most clearly shown in the nuclei of cultured squamous carcinoma cells. Observation by confocal laser scanning microscopy revealed its fine granular distribution in the nucleus but not in the nucleolus. The amount of the antigen as assessed by fluorescence intensity of immunostaining differed among cell lines, but there were no correlation with the stage of differentiation in squamous cell lineage or to the tissues where carcinoma arose. Attempt to reveal ultrastructural localization did not succeed because of the weaknees of the antigenicity againt various fixatives. Since funcitonal analysis indicates the presence of IP_3-R only in the nuclear membrane but not in the nucleus, 10A6 monoclonal antibody might react with crossreacting antigen other than IP_3-R protein itself. Further investigation to reveal nature of the antigen in the nucleus of human cells is now in progress.

肌醇1，4，5-三磷酸(IP_3)是细胞内信号转导的重要信使之一，与其受体(IP_3-R)共同调控细胞内钙离子的流动。Mikoshiba等人已经发现了Ip_3-R蛋白及其编码基因，并制备了与小鼠IP_3-R蛋白反应的单抗。利用这些单抗(10A6、4C11、18A10)与人的交叉反应，我们显示了IP_3-R在人体组织和细胞中的分布，特别是在正常皮肤，包括良性鳞状上皮、鳞状细胞癌组织和鳞癌培养细胞系中。在本研究项目中，我们检测了10A6单抗与细胞核内抗原的特异性反应性。在培养的鳞状癌细胞的细胞核中，该抗原的存在最为明显。激光共聚焦扫描显微镜观察发现，其细小颗粒分布在细胞核内，但不在核仁内。免疫组织化学染色的荧光强度测定的抗原含量在不同的细胞系中不同，但与鳞状细胞系的分化阶段或发生癌的组织无关。试图揭示超微结构的定位没有成功，因为对各种固定剂的抗原性很弱。由于功能分析表明IP_3-R仅存在于核膜上，而不存在于细胞核内，因此，10A6单抗可能与交叉反应的抗原反应，而不是与IP_3-R蛋白本身发生反应。目前正在进行进一步的研究，以揭示人类细胞核中这种抗原的性质。

项目成果

Hiromi Shiba: "The effects of calcium ionophore A23187 and TPA on the expression of IP3-receptor in human B cell lines" Acta Histochemica et cytochemica. 27(4). 387- (1994)

Hiromi Shiba：“钙离子载体 A23187 和 TPA 对人类 B 细胞系中 IP3 受体表达的影响”《组织化学与细胞化学学报》。

Hiromi Shiba: "The effects of calcium ionophore A23187 and TPA on the expression of IP3-receptor in human B cell lines." Acta Histochemica et cytochemica. 27 (4). 387 (1994)

Hiromi Shiba：“钙离子载体 A23187 和 TPA 对人类 B 细胞系中 IP3 受体表达的影响。”

Taiji Katoh: "Immunoreactivity of an anti-mouse inositol 1, 4, 5-trisphosphate receptor monoclonal anitbody with the nuclear antigen of the human cells." Acta Histochemica et cytochemica.27(4). 423- (1994)

Taiji Katoh：“抗小鼠肌醇 1,4,5-三磷酸受体单克隆抗体与人类细胞核抗原的免疫反应性。”

Taiji Katoh: "Immunoreactivity of an antimouse inositol 1,4,5-trisphosphate receptor monoclonal antibody with the nuclear antigen of the human cells." Acta Histochemica et cytochemica. 27 (4). 423 (1994)

Taiji Katoh：“抗小鼠肌醇 1,4,5-三磷酸受体单克隆抗体与人类细胞核抗原的免疫反应性。”