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in situ hybridization study on PTHrP gene expression in oral carcinoma

口腔癌PTHrP基因表达的原位杂交研究

基本信息

批准号：
12671972
负责人：
KATAGIRI Masataka
金额：
$ 1.86万
依托单位：
The Nippon Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671972/
关键词：
parathyroid hormone related protein PTHrP in situ hybridization oral squamous cell carcinoma hypercalcemia immunohistochemistry oral cancer histopathological malignancy in situハイブリダイゼーション

项目摘要

The purpose of this study was to determine whether the expression of mRNA of PTHrP is related to the immunohistochemical stains of PTHrP 1 -34 and keratinization. Biopsy specimens from 33 patients, 11 females and 22 males, with oral squamous cell carcinoma and 4 normal mucosal specimens were used for this study. A previously described method of immunohistochemical staining (LSAB method) with polyclonal 1-34 antibodies and non-isotopic in situ hybridization were performed on 5 μm thickness paraffin sections. We used digoxigenin labeled oligonucleotide probe which was complement to the human PTHrP mRNA coding for the amino acid sequence PTHrP 6-16. Positive signals of PTHrP mRNA were present in 30/33 (91 %) specimens. All specimens were classified in two groups, one showed slight stains and another showed intense staining by the immunohistochemical investigation. Although the percentage (100 %, 13/13) of positive signals of the mRNA in the intense stain group is higher than that (85 %, 17/20) in the weak stain group, it was difficult to distinguish two groups by expression of mRNA. The results suggest that transcription of PTHrP might not affect much to keratinization or differentiation of oral cancer. Post translational processing of PTHrP may take a major role for this phenomenon. Additional study is required to elucidate this issue.Another special attention was focused on the expression of p53 and Ki-67 for comparing PTHrP expression. p53 expression was frequently occurred in squamous cell carcinoma specimens. Though some researchers reported that there was a relationship between p53 mutations and PTHrP in some other malignancies, no significant immunohistochemical relationship between those was revealed in squamous cell carcinoma.

本研究旨在探讨PTHrPmRNA表达与PTHrP 1 - 34免疫组化染色及角化的关系。本研究使用了33例口腔鳞状细胞癌患者（11例女性和22例男性）的活检标本和4例正常粘膜标本。在5 μm厚的石蜡切片上进行先前描述的用多克隆1-34抗体的免疫组织化学染色（LSAB方法）和非同位素原位杂交。我们使用地高辛标记的寡核苷酸探针，其与编码PTHrP 6-16氨基酸序列的人PTHrP mRNA互补。PTHrPmRNA阳性率为91%（30/33）。所有标本经免疫组化染色分为两组，一组为轻度染色，另一组为强染色。强染色组mRNA阳性率（100%，13/13）高于弱染色组（85%，17/20），但很难通过mRNA表达区分两组。结果提示PTHrP的转录对口腔癌的角化和分化无明显影响。PTHrP的翻译后加工可能在这一现象中起主要作用。另一个特别关注的是p53和Ki-67的表达，以比较PTHrP的表达。p53蛋白在鳞状细胞癌组织中的表达率较高。虽然有研究报道在其他一些恶性肿瘤中p53突变与PTHrP之间存在相关性，但在鳞状细胞癌中没有发现它们之间的显著免疫组化关系。