Gene Expression and Development of Odontogenic tumors

牙源性肿瘤的基因表达和发育

基本信息

批准号：
06454511
负责人：
NAGAI Noriyuki
金额：
$ 4.1万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454511/
关键词：
Odontogenic tumors Amelogenin Ameloblastoma Adenoid odontogenic tumors mRNA Digoxigenin Labeling Ameloblast Columnar cells Ameloblastoma エナメル上皮腫 ras蛋白 c-erbB-2蛋白 PCNA p53蛋白悪性エナメル上皮腫 c-myc蛋白免疫組織化学

项目摘要

The classification of odontogenic tumors is related to the differentiation degree of tumor cells. The localization of amelogenin protein and the expression of amelogenin mRNA are among the indexes of odontogenic tumor cell defferentiation. In our study the probes of amelogenin protein and amelogenin mRNA were made and used to observe their distribution in odontogenic tumors.Materials and methods : 1.Normal control : Wistar strain rat embryoes at 20 days after gestation were used for investigation of amelogenin protein and mRNA distribution in tooth germ.The specimens were fixed by 4% paraformaldehyde perfusion. 2.Making of probes : Total RNA was extracted from human plexiform type ameloblastoma. Primers (sense : 5-TCGGTCCTGCCTCAT-CACCATCC-3 ; antisense : 5-CCGCTTGGTCTTGTCTGTTG-3) were used for amplification using PCR method. The amplified cDNA was loaded on Vector (Bluescript) and labeled with Dig RNA Labeling Kit (Boehringer Mannheim) to make Digoxigenin labeled single strand RNA prob … More e by in vitro transcription. 3.Hybridization : The probe was hybridizad with the specimens at 50^0C for 16 hours. Antigen-antibody reaction was performed using Nucleic acid Detection Kit (Boehringer Mannheim) as to evaluate the hybridization under light microscope. In addition, polyclonal anti-amelogenin antibody was used to localize the distribution of amelogenin protein in odontogenic tumors.Results and discussion : Both amelogenin protein and mRNA were positive in rat and mouse incisors but negative in undifferentiated enamel epithelia. Amelogenin mRNA was detected frequently in the cells of the central region of the follicles and usually positive in the cytoplasm of columnar cells near to basement membrane while amelogenin protein was occasionally detected only in a few cases of ameloblastomas. These findins suggest that amelogenin mRNA is generally present in ameloblastomas. Amelogenin mRNA was commonly detected in adenoid ondontogenic tumor cells. In addition the localization of amelogenin protein was also observed in adenoid odontogenic tumors. These results proved at molecular level that adenoid odontogenic tumor is a well differentiated tumor. Apart from the above-mentioned investigations immunohistochemistry was also adopted to evaluate cell proliferation, oncogene product and antioncogene product in ameloblastomas. These immunohistochemical studies indicated that immunodetection of PCNA,c-myc protein, ras protein and p53 protein is helpful in evaluating the malignance of ameloblastomas. Less

牙源性肿瘤的分类与肿瘤细胞的分化程度有关。牙源性蛋白的定位和牙源性肿瘤的表达是牙源性肿瘤细胞脱育的指标之一。在我们的研究中，制作了牙源性蛋白质和牙源性肿瘤的问题，并用于观察它们在牙源性肿瘤中的分布。材料和方法：1。妊娠20天后的wistar菌株大鼠胚胎用于研究Amelogenin蛋白质蛋白和MRNA蛋白质和mRNA分布在牙齿胚芽中。固定在牙齿培养基中。固定了4％para para para perfuly。 2.问题的产生：从人类丛状型蛋白酶瘤中提取总RNA。使用PCR方法，使用引物（有觉：5-TCGGTCCTGCCTCAT-CACCATCC-3；反义：5-CCGCTTGTCTTGTCTGTTG-3）用于扩增使用PCR方法。将放大的cDNA加载在载体（Bluescript）上，并用DIG RNA标记试剂盒（Boehringer Mannheim）标记，以使二高氧素标记为单链RNA概率…通过体外转录更多E。 3.杂交：该探针在50^0c的物种中杂交16小时。使用核酸检测试剂盒（Boehringer Mannheim）进行抗原抗体反应，以评估光学显微镜下的杂交。此外，多克隆抗淀粉蛋白抗体在牙源性肿瘤中定位了氨基苯甲酸蛋白蛋白的分布。分辨和讨论：杏仁蛋白蛋白和小鼠切牙的氨甲贝宁蛋白和mRNA均为阳性，但未分化的搪瓷上皮elia均为阴性。在孔的中央区域的细胞中，经常检测到氨基苯甲酸mRNA，通常在柱状细胞的细胞质中呈阳性，而在基底膜附近，而有时仅在少量amel脂蛋白细胞瘤中仅检测到氨甲型蛋白蛋白。这些发现素表明蛋白细胞母细胞瘤。木母细胞瘤通常在类Ontotogent肿瘤细胞中呈现。另外，在腺样性源基因源性肿瘤中还观察到了肿瘤源性蛋白的定位。这些结果在分子水平上证明，腺样性的牙基因源性肿瘤是一个很好的分化肿瘤。除了上述研究外，还采用了免疫组织化学来评估细胞增殖，癌基因产物和抗四核基因产物。这些免疫组织化学研究表明，PCNA，C-MYC蛋白，RAS蛋白和p53蛋白的免疫检测有助于评估杏仁核细胞瘤的恶性肿瘤。较少的