LACTB基因的可变剪接分析及不同转录本在白血病细胞系中表达的特点

Alternative Splicing Analysis of LACTB Gene and Expression Characteristics of Different Transcripts in Leukemia Cell Lines

目的:检测白血病细胞系中β-内酰胺酶(Lactamaseβ,LACTB)基因不同转录本的表达。方法:使用NCBI网站和DNAstar软件对LACTB进行生物信息学分析。采用反转录PCR(RT-PCR)、DNA测序和克隆测序的方法检测白血病细胞系(THP-1、HL60、K562、U937、Jurkat和Raji)中LACTB不同转录本的表达,采用实时荧光定量PCR方法检测LACTB不同转录本的表达量。结果:LACTB存在多种剪接异构体,并可产生多种氮端保守且碳端不同的蛋白异构体。LACTB在白血病细胞系中存在多种剪接异构体,且不同的细胞系中存在不同的表达模式,包括XR1、V1、V2和V3。总LACTB在HL60细胞中高表达,在Raji细胞中低表达,差异具有统计学意义(P <0.05);V1在U937细胞中高表达,在Raji细胞中低表达,差异具有统计学意义(P <0.05);V2在HL60细胞中高表达,在Raji细胞中低表达,差异具有统计学意义(P <0.05);V3在THP-1细胞中低表达,与正常骨髓相比差异具有统计学意义(P <0.05)。结论:LACTB在白血病细胞系中存在多种剪接异构体,且不同的细胞系中存在不同的表达模式。

Objective: To detect the expression of different transcripts of lactamase β( LACTB) gene in leukemic cell lines. Methods: NCBI website and DNAstar software were used to detect the Bioinformatics analysis of LACTB. The expression of different transcripts of LACTB gene in leukemic cell lines( THP-1,HL60,K562,U937,Jurkat and Raji)was detected by reverse transcription PCR( RT-PCR),DNA and clone sequencing;the expression of different transcripts of LACTB gene in leukemic cell lines was detected by Quantitative Real-time PCR. Results: There were a variety of splicing isomers in LACTB,and it could produce a variety of protein isomers with conserved N-terminal and different C-terminal,moreover,there were many splice isoforms of LACTB in leukemia cell lines,and there were different expression patterns in different cell lines,including XR1,V1,V2 and V3. The expression of total LACTB showed high in HL60 cells,while low in Raji cells,and the difference was statistically significant( P < 0. 05). The V1 was high expression in U937 cells but low in Raji cells,and the difference was statistically significant( P < 0. 05). V2 was high expression in HL60 cells but lowly in Raji cells,and the difference was statistically significant( P < 0. 05).The expression of V3 was low in THP-1 cells,which was significantly different as compared with that in normal bone marrow( P < 0. 05). Conclusion: The reaserch found that there are many splice isomers of LACTB in leukemic cell lines,and there are different expression patterns in different cell lines.