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目的检测肺癌组织中黑色素瘤抗原-3基因(MAGE-A3)mRNA的表达。方法用逆转录-套式聚合酶链反应(RT-PCR)对31例肺癌患者癌组织和相应癌旁组织MAGE-A3mRNA表达情况进行测定;PE-377DNA测序仪对5例10个RT-PCR扩增产物中的目的基因片段进行DNA序列测定。结果31例肺癌患者癌组织中26例表达MAGE-A3mRNA,阳性率为83.9%;相应的癌旁组织均未表达。DNA序列测定证明PCR扩增产物中目的基因片段均为MAGE-A3cDNA序列,所测5例样本中4例样本有两个相同位置的碱基发生了点突变(C2773→T2773;G2807→A2807),导致一个氨基酸残基改变(E143→K)。结论MAGE-A3mRNA在肺癌中呈高比例表达,提示此抗原有可能作为肺癌患者免疫治疗的靶抗原。我国肺癌患者中存在MAGE-A3基因个别位点的变异。
Objective To detect the expression of melanoma antigen-3 (MAGE-A3) mRNA in lung cancer. Methods The expression of MAGE-A3 mRNA in 31 cases of lung cancer and adjacent normal tissues was detected by reverse transcription-nested polymerase chain reaction (RT-PCR). Five cases of 10 cases of RT-PCR amplification The amplified product of the target gene fragment DNA sequence determination. Results MAGE-A3 mRNA was expressed in 26 of 31 lung cancer patients, the positive rate was 83.9%. The corresponding paracancerous tissues were not expressed. DNA sequencing confirmed that the target gene fragments in PCR amplification products were all MAGE-A3cDNA sequences. There were two point mutations (C2773 → T2773; G2807 → A2807) in two of the four samples in the 5 samples tested, Resulting in a change of one amino acid residue (E143 → K). Conclusion MAGE-A3 mRNA is highly expressed in lung cancer, suggesting that this antigen might be the target antigen for immunotherapy in patients with lung cancer. The mutation of MAGE-A3 gene in lung cancer patients in our country exists.