Wang Lei, Zhu Hong-guang
Department of Pathology, Shanghai Medical College, Fudan University and Pathology Center, Institute of Biomedical Sciences, Fudan University, Shanghai 200032, China.
Zhonghua Bing Li Xue Za Zhi. 2006 Apr;35(4):224-7.
To study the clonality of palmar fibromatosis by molecular genetic analysis of X chromosome inactivation pattern at a polymorphic site of human androgen receptor gene (HUMARA).
Twelve female cases of palmar fibromatosis were enrolled into this study. Hematoxylin and eosin-stained sections of paraffin-embedded, formalin-fixed tissues were microdissected by laser capture microdissection technology in order to obtain the proliferative spindle cells. Tumor cells isolated from rectal adenocarcinoma in a female patient were used as positive control. The genomic DNAs were extracted with phenol and chloroform, digested with methylation-sensitive restriction endonuclease HpaII, and amplified by polymerase chain reaction (PCR) using primers targeted to a highly polymorphic short tandem repeat of HUMARA. The amplimers were separated on vertical 8% non-denaturing polyacrylamide gels and the patterns were visualized with ethidium bromide stain.
The methodology for clonality analysis was validated in the positive control using rectal adenocarcinoma cells. Among the 12 cases studied, PCR amplification failed in 3 samples and 1 sample showed homozygosity which was not suitable for further analysis. Eight samples were successfully amplified and showed a random X chromosome inactivation pattern, suggesting polyclonality in these lesions.
Palmar fibromatosis is a polyclonal condition and should be considered as a form of non-neoplasmic fibroblastic proliferation.
通过对人类雄激素受体基因(HUMARA)多态性位点的X染色体失活模式进行分子遗传学分析,研究掌纤维瘤病的克隆性。
本研究纳入12例女性掌纤维瘤病患者。采用激光捕获显微切割技术对福尔马林固定、石蜡包埋组织的苏木精-伊红染色切片进行显微切割,以获取增殖的梭形细胞。以一名女性患者直肠腺癌分离的肿瘤细胞作为阳性对照。用酚和氯仿提取基因组DNA,用甲基化敏感限制性内切酶HpaII消化,并用针对HUMARA高度多态性短串联重复序列的引物进行聚合酶链反应(PCR)扩增。扩增产物在8%垂直非变性聚丙烯酰胺凝胶上分离,并用溴化乙锭染色观察条带模式。
在使用直肠腺癌细胞的阳性对照中验证了克隆性分析方法。在研究的12例病例中,3个样本PCR扩增失败,1个样本显示纯合性,不适于进一步分析。8个样本成功扩增,显示随机X染色体失活模式,提示这些病变为多克隆性。
掌纤维瘤病是一种多克隆性疾病,应被视为非肿瘤性成纤维细胞增殖的一种形式。