摘要
目的 根据女性细胞中两条 X染色体上甘油磷酸激酶 (PGK)基因多态性 ,建立克隆性分析技术 ,并探讨子宫平滑肌瘤的克隆性 .方法 从新鲜子宫平滑肌瘤组织中提取DNA,用限制性内切酶 Hpa II消化样品后 ,PCR扩增 ,产物再经 Bst XI内切酶消化 ,琼脂糖凝胶电泳显示结果 .结果 在扩增成功的 32例标本中 ,10例 (31% )具有 Bst XI酶切位点的多态性 .其中 1例 X染色体失活发生了严重的偏移 ,不适合作克隆性分析 .对其中 7例标本、15个肿瘤结节进行了克隆性分析 .结果显示 ,7例中 15个肿瘤结节全部为单克隆性起源 ,其中 2例 (分别有 2个肌瘤 )的肿瘤结节起源于不同的克隆 ,1例中相邻的 6个肌瘤结节具有相同的酶切带型 ,提示不同肿瘤结节起源于同一克隆 .结论 1运用 PGK克隆性分析技术有助于局灶性或结节性病变性质的确定 ;2子宫平滑肌瘤是单克隆细胞群体 ;多发性平滑肌瘤中 ,多数病例不同肿瘤结节相互独立 。
AIM To establish clonality analysis based on the random inactivation of X chromosome and the heterozygosity in female somatic cells at the phosphoglycerate kinase (PGK) locus, and to study the clonal composition of uterine leiomyomas. METHODS Genomic DNA was extracted from uterine leiomyoma samples, digested with restriction enzyme Hpa II and amplified via PCR. The products were then treated with polymorphic restriction enzyme Bst XI, and resolved through agarose gel electrophoresis. RESULTS Ten cases were shown to carry polymorphic Bst XI site in the PGK locus in 32 cases of samples which were amplified successfully. One of the 10 cases showed marked skewing of the X chromosome inactivation, and was excluded from the assay. Fifteen tumors from 7 cases were then subjected to the analysis. All of the tumors showed clonal proliferation. The relationship among multiple tumors was also determined by comparison of their inactivated alleles. In 2 cases with 2 tumors which were located far from each other, inactivation if different alleles were observed, indicating that these nodules are indeed independent. However, an identical inactivated allele was found in one case among six tumor nodules, strongly suggesting a unifocal origin of all of these nodules. CONCLUSION ① Clonality analysis is a useful approach for a better understanding of nodular or focal lesions with an uncertain biological behavior. ② Uterine leiomyomas are clonal in cell compo sition. Among the multiple nodule cases, different tumors are independent in most cases, but may be of unicellular origin in some cases.
出处
《第四军医大学学报》
北大核心
2001年第17期1576-1582,共7页
Journal of the Fourth Military Medical University
关键词
克隆
甘油激酶
子宫平滑肌瘤
cloning, molecular
glycerol kinase
smooth muscle neoplasm, uterus
