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目的应用多重PCR方法扩增TCRα链全长序列,分析乳腺癌浸润淋巴结αT细胞受体谱型偏移。方法选取乳腺癌转移淋巴结,提取RNA后逆转录,多重PCR扩增TCRα链全长序列,构建重组质粒并测序,利用网上TCR资源分析序列。结果 2例患者各扩增出3个α链序列。乳腺癌TCRα限制性取用AV1、AV12亚家族。CDR3区序列分析增生T细胞克隆具有不同的氨基酸序列。结论乳腺癌浸润淋巴结扩增的TCR家族存在限制性取用,克隆性增生T细胞CDR3序列不同。
Objective To amplify the full-length sequence of TCRα chain by multiplex PCR and analyze the shift of receptor type of αT cells in infiltrating lymph node of breast cancer. Methods The lymph node metastasis of breast cancer was selected, RNA was extracted and reverse transcribed. The full length of TCRα chain was amplified by multiplex PCR. The recombinant plasmid was constructed and sequenced. The sequence was analyzed by online TCR. Results Two α-chain sequences were amplified in two patients. Breast cancer TCRα restrictive access AV1, AV12 subfamily. CDR3 Region Sequence Analysis The proliferating T cell clones have different amino acid sequences. Conclusions The TCR family amplified by infiltrating lymph nodes in breast cancer has restricted access, and CDR3 sequences of clonal proliferating T cells are different.