MLPA技术在检测胰腺癌细胞系9p21大片段缺失中的应用

目的:评价MLPA技术检测染色体9p21区大片段缺失的有效性及判断缺失边界的准确性,并探讨染色体9p21区大片段缺失的意义.方法:利用MLPA技术检测胰腺癌细胞系PANC-1和BxPC3的9p21区缺失情况,并利用常规PCR实验对MLPA缺失边界进行验证.结果:MLPA检测发现PANC-1和BxPC3均存在累及MTAP、CDKN2A和CDKN2B等基因大片段缺失的情况.PCR实验证实MLPA对PANC-1端粒侧的缺失边界判断准确.PANC-1细胞系染色体9p21区大片段缺失的端粒侧缺失断点位于MTAP基因内.结论:MLPA技术是检测染色体9p21大片段缺失的有效方法,能准确判断大片段缺失边界范围,适合于aCGH检测后大样本筛查.PANC-1细胞系中染色体9p21区大片段缺失可能改变MTAP基因结构.

Application of multiplex ligation-based probe amplification in detection of large deletions on chromosome 9p21 in pancreatic cancer cell lines

Objective:To evaluate the efficiency and accuracy of multiplex ligation-based probe amplification (MLPA) method in detection of large deletions and their borders on chromosome 9p21 as well as the significance of these large deletions.Methods:MLPA was used to detect large deletions in pancreatic cell lines PANC-1 and BxPC3.Conventional PCR was used to confirm distal borders of large deletions.Results:Large deletions involved in MTAP,CDKN2A and CDKN2B were found to exist in these two pancreatic cancer cell lines by MLPA.Conventional PCR confirmed the accuracy in detection of deletion borders by MLAP.Moreover,in PANC-1,the telomeric border of large deletion was located within MTAP.Conclusions:MLPA is an efficient way for detection of large deletions and their borders on chromosome 9p21,which is suitable for deletion screening in large sample sizes after array-based comparative genomic hybridization (aCGH) study.In PANC-1,MTAP gene structure may be altered by chromosome 9p21 deletion.