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应用聚合酶链反应-单链构象多态性分析技术检测胃肠道淋巴瘤中T细胞受体γ基因重排

Investigation of T-cell receptor-gamma gene rearrangement in gastrointestinal lymphomas by PCR-SSCP analysis.

作者信息

Han Xi-Qun, He Li, Shong Lan-Ying, Jiang Hui-Yong, Zhu Mei-Gang, Zhao Tong

机构信息

Department of Pathology, the First Military Medical University, Guangzhou 510515, Guangdong Province, China.

出版信息

World J Gastroenterol. 2004 Apr 1;10(7):995-9. doi: 10.3748/wjg.v10.i7.995.

Abstract

AIM

To analyze the characterization of T-cell receptor-gamma (TCR-gamma) gene rearrangement in the gastrointestinal lymphomas and evaluate the value of PCR-SSCP analysis in gastrointestinal lymphomas investigation.

METHODS

TCR-gamma gene rearrangement segments of gastrointestinal lymphomas were cloned and sequenced. Single clone plasmid and mixed clone plasmids were subsequently submitted to PCR-SSCP analysis to investigate the relationship between the number of amplified clones and band patterns of the amplified products. The PCR products of TCR-gamma gene rearrangement of 40 gastrointestinal lymphomas were electrophoresed on agarose gels and the positive cases on agarose gels were studied by SSCP analysis.

RESULTS

The sequencing showed that TCR-gamma gene rearrangement of the gastrointestinal lymphomas included functional gene and pseudogene with extensive variety in the junctional regions. In SSCP analysis, the number of the single-stranded bands was about two times of the number of amplified clones, and double-stranded band became broad with the increased number of the amplified clones. Thirteen of the 25 B-cell gastrointestinal lymphomas and 14 of the 15 gastrointestinal T-cell lymphomas were positive detected on agarose gel electrophoresis. Of the positive cases detected by SSCP analysis, 3 B-cell lymphomas and 13 T-cell lymphomas showed positive bands. The other cases showed only smears. The rearranged pattern included 13 monoallelic gene rearrangements and 3 biallelic or oligoclonal gene rearrangements.

CONCLUSION

The pattern of TCR-gamma gene rearrangement in gastrointestinal lymphomas are similar to that of the nodular lymphomas. PCR-SSCP analysis for TCR-gamma gene rearrangement can be applied both for adjuvant diagnosis of gastrointestinal lymphomas and analysis of the gene rearrangement pattern. The ratio of TCR-gamma gene rearrangements occurred in T-cell gastrointestinal lymphomas is significantly higher than that in B-cell gastrointestinal lymphomas. The gene rearrangement pattern involves monoallelic and biallelic (or oligoclonal) gene rearrangement.

摘要

目的

分析胃肠道淋巴瘤中T细胞受体γ(TCR-γ)基因重排的特征,并评估聚合酶链反应-单链构象多态性分析(PCR-SSCP)在胃肠道淋巴瘤研究中的价值。

方法

对胃肠道淋巴瘤的TCR-γ基因重排片段进行克隆和测序。随后将单克隆质粒和混合克隆质粒进行PCR-SSCP分析,以研究扩增克隆数量与扩增产物条带模式之间的关系。对40例胃肠道淋巴瘤的TCR-γ基因重排的PCR产物进行琼脂糖凝胶电泳,并对琼脂糖凝胶上的阳性病例进行SSCP分析。

结果

测序显示胃肠道淋巴瘤的TCR-γ基因重排包括功能性基因和假基因,在连接区具有广泛的多样性。在SSCP分析中,单链带的数量约为扩增克隆数量的两倍,双链带随着扩增克隆数量的增加而变宽。25例B细胞胃肠道淋巴瘤中有13例,15例胃肠道T细胞淋巴瘤中有14例在琼脂糖凝胶电泳上检测为阳性。在SSCP分析检测为阳性的病例中,3例B细胞淋巴瘤和13例T细胞淋巴瘤显示出阳性条带。其他病例仅显示涂片。重排模式包括13种单等位基因重排和3种双等位基因或寡克隆基因重排。

结论

胃肠道淋巴瘤中TCR-γ基因重排模式与结节性淋巴瘤相似。PCR-SSCP分析TCR-γ基因重排可用于胃肠道淋巴瘤的辅助诊断和基因重排模式分析。TCR-γ基因重排在胃肠道T细胞淋巴瘤中的发生率明显高于B细胞胃肠道淋巴瘤。基因重排模式涉及单等位基因和双等位基因(或寡克隆)重排。

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