Analysis of clonality in odontogenic tumors by PCR assay of X chromosome gene

成釉细胞瘤是最常见的牙源性肿瘤，被归类为良性肿瘤。但表现为浸润性生长，破坏周围骨结构或软组织。基因组技术的最新进展使通过X染色体连锁基因的PCR分析来评估病变的克隆性成为可能。本研究以9例女性成釉细胞瘤(均为滤泡型)为研究对象，采用人雄激素受体基因多态性的聚合酶链式反应分析方法，对成釉细胞瘤的克隆性进行研究。将福尔马林固定、石蜡包埋的组织切片，分离肿瘤区域。以HpaII酶切产物和未消化的DNA为模板，扩增包括三核苷酸重复序列在内的雄激素受体基因外显子1。将聚合酶链式反应产物负载于聚丙烯酰胺凝胶上，进行凝胶电泳。凝胶银染，9例中有3例无多态等位基因。其余6例在凝胶上显示两条带，因此被判定为多克隆。

Ameloblastoma, the most popular odontogenic tumor, is classified as benign tumor. However, it shows infiltrative growth and destroy surrounding bony structure or soft tissues. Namely, "generally benign, but locally aggressive".Recent advances in genomic technology enable the assesment of clonality in a lesion by PCR assay of X chromosome-linked gene. The aim of this study is to evaluate the clonality in ameloblastomas by PCR assay of human androgen receptor gene polymorphism.From tissue-archive in our institute, 9 cases of ameloblastoma (all follicular type) arisen in female were selected as the specimens. Formalin-fixed, paraffin-embedded tissues were sliced and tumor area was isolated. The specimens were digested with proteinase-K and a part of them were digested with HpaII.The HpaII digested product and remaining undigested DNA were used as templates for PCR amplification for androgen receptor gene exonl including polymorphic trinucleotide repeat. The PCR product was loaded on polyacrylamide gel and phoresed. The gel was stained with silver staining.Three of 9 cases did not have polymorhic allele. Remaining 6 cases showed two bands on the gel so that were evaluated as polyclonal.