原发性探索结直肠癌患者外周血和组织中 CDR3 谱型和 TCR β 链的分子特征。

Primary exploration of CDR3 spectratyping and molecular features of TCR β chain in the peripheral blood and tissue of patients with colorectal carcinoma.

机构信息

Clinic Laboratory, The Affiliated Hospital of Jining Medical College, Jining, China.

出版信息

Cancer Epidemiol. 2010 Dec;34(6):733-40. doi: 10.1016/j.canep.2010.06.010. Epub 2010 Jul 31.

DOI:10.1016/j.canep.2010.06.010

PMID:20674537

Abstract

BACKGROUND

The immunoscope spectratyping to TRBV CDR3 had been applied in infectious diseases, tumors, autoimmune diseases and so on, this study aimed to primarily explore CDR3 spectratyping and molecular features of TCR β chain in the peripheral blood and tissue of patients with colorectal carcinoma.

METHODS

Blood and tissue samples were collected from seven patients with colorectal carcinoma (CRC), while blood samples were also collected from two healthy controls as control. Using the real-time fluorescence quantitative reverse transcription polymerase chain reaction (RQ-PCR) and DNA melting curve analysis techniques, the features of T-cell receptor beta chain variable region (TRBV) were determined.

RESULTS

The gene melting spectral pattern (GMSP) of 24 TRBV gene families exhibited a highly diverse multimodal shape for most of the TRBV gene families, compared to healthy controls, the more GMSP of patients with CRC showed either a single peak, or several prominent melting peaks (skewed) for certain TRBV gene families. Different patients have different skewed patterns. In the analysis results of sequence, some TRBV CDR3 gene families showed sharing the same motif, such as, TRBV6 of P5, TRBV13.1 of P6 and TRBV21 of P7 (tissue sample) shared the same motif 'GT'; TRBV1 of P1 and TRBV21 of P7 shared the same motif 'AGG', TRBV11 of P1, TRBV21 of P5 and TRBV21 of P7 (tissue sample) shared the same motif 'TDTQY', and even TRBV21 of P5 and P7 (tissue sample) shared the large motif 'SGTDTQY'. As a whole, most of TRBV gene families have the similar motifs 'X-Q', the nucleotide 'X' mainly was 'E', it also was possible be 'T', 'G' or 'K' in some CDR3 gene families of patients with CRC.

CONCLUSIONS

There were different GMSPs in different patients with CRC, CDR3 spectratyping and the molecular features of TCR β chain in the peripheral blood and tissue of patients with CRC were not same or similar, this information would provide ideas for individualized therapy to CRC.

摘要

背景

免疫谱分析 TRBV CDR3 已应用于传染病、肿瘤、自身免疫性疾病等领域，本研究旨在初步探讨结直肠癌患者外周血和组织中 TCRβ 链 CDR3 谱分析和分子特征。

方法

收集 7 例结直肠癌（CRC）患者的血液和组织样本，同时收集 2 例健康对照者的血液样本作为对照。采用实时荧光定量逆转录聚合酶链反应（RQ-PCR）和 DNA 熔解曲线分析技术，确定 T 细胞受体β链可变区（TRBV）的特征。

结果

24 个 TRBV 基因家族的基因熔解光谱模式（GMSP）呈现出高度多样化的多峰形状，大多数 TRBV 基因家族与健康对照组相比，CRC 患者的 GMSP 要么呈现单个峰，要么呈现几个突出的熔解峰（偏斜），对于某些 TRBV 基因家族。不同患者的偏斜模式不同。在序列分析结果中，一些 TRBV CDR3 基因家族显示出相同的基序，例如，P5 的 TRBV6、P6 的 TRBV13.1 和 P7 的 TRBV21（组织样本）共享相同的基序“GT”；P1 的 TRBV1 和 P7 的 TRBV21 共享相同的基序“AGG”，P1 的 TRBV11、P5 的 TRBV21 和 P7（组织样本）共享相同的基序“TDTQY”，甚至 P5 和 P7（组织样本）的 TRBV21 共享大基序“SGTDTQY”。总体而言，大多数 TRBV 基因家族具有相似的基序“X-Q”，核苷酸“X”主要是“E”，在 CRC 患者的一些 CDR3 基因家族中也可能是“T”、“G”或“K”。