摘要
目的:比较正常人和B细胞白血病患者外周血中黏膜相关淋巴样组织淋巴瘤转位基因1(MALT1)2种异构体的分布和表达差异。方法:根据MALT1基因的结构特点,设计2对引物,通过建立2种转录本的逆转录聚合酶链式反应(RT-PCR)方法分析8例B细胞急性淋巴细胞白血病(B-ALL)、8例B细胞慢性淋巴细胞白血病(B-CLL)和8例正常人的外周血单个核细胞(PBMC)中MALT1的表达情况。实时定量PCR分析各组样本的MALT1 mRNA表达水平。结果:所有样本中均能得到所预期的PCR产物,其中第1对引物可以扩增出2条片段,分别为MALT1转录本1(包含第5、6、7、8、9外显子)和MALT1转录本2(包含第5、6、8、9外显子),第2对引物只能扩增出1条包含转录本1的片段,所有PCR产物均通过核苷酸序列分析证实。MALT1 2种转录本均在外周血中有表达,并且MALT1转录本2表达水平高于转录本1。通过灰度分析发现,在B-ALL中,MALT1转录本1的灰度与正常人相比显著增高(P<0.05),而MALT1转录本2显著降低(P<0.05);在B-CLL中,MALT1 2种转录本的灰度与正常人相比无显著差异(P>0.05)。此外,B-ALL中,MALT1 mRNA表达水平(中位数:1.253)低于正常人(中位数:1.976)(P=0.05);而B-CLL中,MALT1 mRNA的表达水平与正常人相比无显著差异(P>0.05)。结论:MALT1 2种转录本均表达于正常和B细胞白血病样本中,但B-ALL中两者的表达水平有所差异,同时,B-ALL的MALT1 mRNA表达水平也有所降低。B-ALL中MALT1基因表达的异常可能影响MALT1信号通路而与疾病的发生发展相关。
AIM： To investigate the difference of the distribution and expression of mucosa-associated lymphoid tissue lymphoma translocation gene 1 （MALT1） transcripts in healthy individuals and B-cell leukemia patients. METH- ODS： Two pairs of primers were designed according to the sequences of different MALT1 transcripts. RT-PCR was per- formed to detect the distribution of MALT1 gene in peripheral blood mononuelear cells （PBMC） from 8 healthy individuals, 8 patients with B-cell acute lymphocytic leukemia （B-ALL） and 8 patients with B-ceU chronic lymphocytic leukemia （B- CLL）. The mRNA expression of MALT1 in all samples from the 3 groups was determined by real-time PCR. RESULTS： Expected amplicons were amplified by the 2 primer pairs, and 2 PCR products of different sizes were amplified by the first primer pair （ the larger one included exons 5, 6, 7, 8 and 9, and the small one included exons 5, 6, 8 and 9）. Moreover, the MALT1 transcript variant 1 was conformed by the second primer pair. All the PCR products were confirmed by the nu-cleotide sequencing analysis. Two transcripts of MALT! were identified in all samples from both healthy individuals and B- cell leukemia patients, and the expression level of MALT1 was predominant in transcript variant 2. The gray level of MALT! transcript variant 1 in B-ALL patients was significantly higher than that in healthy individuals, whereas the MALT1 tran- script variant 2 was significantly lower than that in healthy group. However, no significant difference of the gray level was found in the 2 MALT! transcript variants in B-CLL group as compared with healthy group. In addition, the expression level of MALT1 mRNA in B-ALL group （ median = 1. 253 ） was lower than that in healthy group （ median = 1. 976, P = 0.05 ）, while no significant difference of the expression level of MALT1 mRNA between B-CLL patients and healthy individuals was found. CONCLUSION： Two transcripts of MALT1 have common distribution in both healthy individuals and B-cell leuke- mia patients, while the significant differences of distribution of MALT1 transcript variants and the expression level of MALT1 mRNA are characterized in B-ALL patients. The abnormal expression of MALT1 gene might involve in the genesis and development of leukemia via MALT1 signaling pathway.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第6期1124-1128,1132,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.91129720)
广东省科技计划(国际合作)项目(No.2012B050600023)
关键词
黏膜相关淋巴样组织淋巴瘤转位基因1
转录本
白血病
B细胞
基因表达
Mucosa-associated lymphoid tissue lymphoma translocation gene 1
Transcript variants
Leukemia, B-cell
Gene expression
