竞争性RT-PCR法定量分析骨巨细胞瘤中TGFβ-1 mRNA的表达水平  

Quantitative determination of TGFβ-1 mRNA in giant cell tumor by competitive polymerase chain reaction

作　　者：刘光耀[1] 孙庆[1] 高忠礼[1] 

机构地区：[1]吉林大学中日联谊医院骨科,吉林长春130031

出　　处：《中国实验诊断学》2001年第2期69-71,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的 用竞争性PCR法检测骨巨细胞瘤（GCT）中TGFβ- 1mRNA，分析其对肿瘤预后的意义。方法 通过RT- PCR法制备内标和外标，将等量的内标和等比稀释的外标进行竞争性共扩增，得到标准曲线，然后将等量的内标和cDNA标本共扩增，通过标准曲线求出标本中TGFβ- 1mRNA的水平，并与GCT分级进行对比。结果 内标和外标在共扩增中表现出很强的竞争性。GCT中TGFβ- 1 mRNA的表达水平随着肿瘤分级的上升明显增加，局部复发和肺转移的标本中TGFβ- 1mRNA表达水平较未复发的标本高。结论 竞争性RT- PCR是一种灵敏，简便、快速检测GCT中TGFβ- 1mRNA表达水平的方法。应用该方法和肿瘤病理分级标准来综合判断肿瘤的预后，有着重要的临床意义。Objective Quantitatively determining TGFβ- 1 mRNA in giant cell tumor by competitive polymerase chain reaction and analyzing the significance of it in estimation of the prognosis of giant cell tumor. Methods The internal standard and external standard were prepared by RT- PCR;the standard curve was acquired by competitive polymerase chain reaction using the fix quantity internal standard and the geometric proportion diluent external standard. The fix quantity internal standard and the cDNA sample were co-expanded. The expression level of TGFβ- 1mRNA of sample was achieved by the standard curve. The significance of it in estimation of the prognosis of giant cell tumor was determined by contrast with the GCT classification. Results The expression level of TGFβ- 1mRNA of GCT increased with the increase of GCT classification. The expression levels of TGFβ- 1mRNA of GCT in local recurrence and lung metastasis samples were higher than those of non recurrence samples. Conclusion Competitive polymerase chain reaction is a kind of method which can examine the expression level of TGFβ- 1mRNA of GCT effectively ，simply and fast. It has important clinic significance in judgement of the prognosis of GCT by the expression level of TGFβ 1mRNA and GCT classification.

关 键 词：竞争性RT-PCR 骨巨细胞瘤 转化生长因子Β-1 TBFβ-1 MRNA 

分 类 号：R738.1[医药卫生—肿瘤]