喉癌组织中表皮生长因子受体mRNA含量测定

Quantitative analysis of epidermal growth factor receptor mRNA in laryngeal carcinoma by reverse transcription-real time polymerase chain reaction with TagMan probe

目的建立 TagMan 探针逆转录一实时荧光定量 PCR 技术测定喉癌组织中表皮生长因子受体(epidermal growth factor receptor,EGFR)mRNA 水平的方法,分析其应用价值。方法设计高特异性引物和探针,以 TagMan 探针逆转录-实时荧光定量 PCR 技术对32例喉鳞状细胞癌组织和癌旁组织学正常喉黏膜组织 EGFR mRNA 表达水平进行测定。构建含有 EGFR 的重组质粒,进行测序鉴定。结果经测序分析证实,含 EGFR 的重组质粒构建成功,喉癌组织中 EGFR mRNA 相对含量(中位数0.025)高于癌旁正常喉黏膜组织中 EGFR mRNA 相对含量(中位数0.008),两者间的差异经Wilcoxon 秩和检验,差异有统计学意义(P<0.01)。结论 TagMan 探针逆转录-实时荧光定量 PCR 技术可作为一种喉癌组织中 EGFR mRNA 水平定量测定的新方法。

Objective To establish a method of quantitative analysis of epidermal growth factor receptor（EGFR） mRNA in laryngeal carcinoma by reverse transcription-real time polymerase chain reaction with TagMan probe and evaluate its applicable value. Methods The technique of reverse transcription-real time polymerase chain reaction with highly specific primers and probe was applied. And with GAPDH as internal standard, EGFR mRNA in 32 laryngeal carcinoma tissues, compared with their own macroscopically normal laryngeal mucosa tissues adjacent to the tumors, were quantitively examined. The recombinant plasmid of EGFR was constructed, and then sequenced. Results The recombinant plasmid of EGFR was successfully constructed. And the mRNA index of EGFR in 32 laryngeal carcinoma tissues in terms of M （QR） was 0. 025 （0. 076 ）, which was higher than 0.008 （0. 027 ）in their own macroscopically normal laryngeal mucosa tissues adjacent to the tumors, P value is 0. 007. Conclusions The method of reverse transcription-real time polymerase chain reaction with TagMan probe is quite objective, precise and high throughput in the application for the mensuration of EGFR mRNA in laryngeal carcinoma.