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喉鳞状细胞癌中EphB4基因突变与表达的研究

Mutation and Expression of EphB4 Gene in Human Laryngeal Squamous Cell Carcinoma
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摘要 目的研究EphB4基因在喉鳞状细胞癌(喉癌)发生、发展中的生物学功能。方法应用RT-PCR和Westernblot的方法,以癌旁组织为参照检测EphB4基因在喉癌组织中的表达情况;应用差异聚合酶链反应(PCR)方法检测肿瘤组织中EphB4基因的扩增情况,聚合酶链反应-低离子浓度-单链构象多态性(PCR-LIS-SSCP)技术结合DNA测序筛查喉癌患者EphB4基因第13、14外显子的突变情况。结果EphB4基因mRNA和蛋白质在喉癌中的表达水平均明显高于癌旁组织(分别为0.738±0.137对0.450±0.173和0.757±0.174对0.509±0.168,均P<0.01),且EphB4基因的高表达与肿瘤的临床分期、分级、肿瘤的直径大小以及肿瘤的转移密切相关,40对配对样本中未检测出基因的扩增和突变。结论EphB4基因的过表达可能与喉癌的发生、发展有关,可能作为喉癌诊断和治疗的一个靶点。 Objective To determine the biological role of EphB4 in laryngeal squamous cell carcinoma (LSCC). Methods The protein and messenger RNA (mRNA) levels of EphBd were determined with RT-PCR and Western blot,respectively,in 40 LSCC and 40 paired adjacent normal tissue samples. Polymerase chain reaction low ionic strength single-stxanded conformation polymorphism (PCR-LIS-SSCP) and DNA sequencing were used to detect the mutation of exon 13 and 14 of EpbBd gene. EpbB4 DNA level was evaluated by differential PCR with β-actin as internal control. Results EpfiB4 expression was detected in all tumor specimens. There was a statistically significantly higher mean EphBd mRNA and protein expression in LSCC specimens than paired adjacent normal tissue specimens. Patients with higher EphB4 mRNA expression level had a higher protein level compared with those with normal EphB4 mRNA expression. In 40 LSCC and 40 paired adjacent normal tissue samples,we did not detect the mutation of exonl3 and 14. No amplification was found in the samples we detected. Conclusion These data are the first to demonstrate the association of EphB4 with LSCC and hence provide a strong rationale for targeting EpfiB4 for LSCC therapies.
出处 《中国医科大学学报》 CAS CSCD 北大核心 2008年第6期733-735,752,共4页 Journal of China Medical University
基金 辽宁省自然科学基金资助项目(2001101039)
关键词 EphB4基因 逆转录项-聚合酶链反应 聚合酶链反应-低离子浓度-单链构象多态性 喉鳞状细胞癌 EphB4 gene RT-PCR PCR-LIS-SSCP LSCC laryngeal squamous cell carcinoma
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