人多发性骨髓瘤细胞系与初治多发性骨髓瘤遗传学异常的比较研究  

Comparative Study of Genetic Aberrations in Human Multiple Myeloma Cell Lines and Newly Diagnosed MM by Fluorescence In Situ Hybridization

作　　者：安刚 谢振卿 李长虹 李倩 易树华 邱录贵 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院,天津300020

出　　处：《中国实验血液学杂志》2010年第6期1505-1510,共6页Journal of Experimental Hematology

基　　金：天津市科技支撑计划重点项目(编号09ZCGYSF1000);卫生部临床科学重点项目基金

摘　　要：本研究采用荧光原位杂交(fluorescence in situ hybridization,FISH)检测人多发性骨髓瘤细胞系(human multiple myeloma cell lines,HMCL)和初治多发性骨髓瘤(MM)的分子遗传学异常并进行比较分析,以帮助筛选浆细胞肿瘤高危遗传学异常,为多发性骨髓瘤MM的危险度分层提供依据,并为利用HMCL进行MM研究提供遗传学资料。选用13q14(RB-1)、14q32(IGHC/IGHV)、1q12(CEP1)、17p13(TP53)荧光探针应用直接法检测7株HMCL,用CD138免疫磁珠分选(magnetic-activated cell sorting,MACS)联合FISH方法检测85例初治MM患者作为对照。对于存在14q32异常的患者进行IGH/CCND1、IGH/FGFR3、IGH/MAF双色双融合探针杂交以检测t(11;14)(q13;q32)、t(4;14)(p16;q32)、(14;16)(q32;q23)。结果显示:7株HMCL中RB-1/D13S19缺失6株(85.7%),P53缺失5株(71%),1q21扩增6株(85.7%);5株(71.4%)HMCL存在IGH异常,IGH/CCND1阳性细胞系1株,IGH/FGFR3阳性细胞系2株,IGH/MAF阳性细胞系3株;其中KMS11细胞系同时存在IGH/FGFR3和IGH/MAF两种异常。85例初治MM细胞遗传学总体检出率为85.9%,38例(44.7%)伴有RB-1缺失,17例(20%)出现p53缺失,45例(52.9%)伴有1q21扩增,53例(62.4%)存在IGH异常,其中IGH/CCND1阳性23例(27.1%),IGH/FGFR3阳性21例(24.7%),IGH/MAF阳性3例(3.5%)。与初治MM相比较,HMCL的抑癌基因p53缺失率和IGH/MAF明显升高(p=0.008,p=0.005),而其他分子遗传学异常检出率未达统计学差异。结论:HMCL作为浆细胞肿瘤恶性程度最高的形式,积累了大量的遗传学异常,p53缺失是其显著特征。绝大多数HMCL来源于疾病晚期的髓外浆细胞肿瘤细胞或者浆细胞白血病,提示p53缺失是这部分极高危浆细胞肿瘤患者的特征。Multiple myeloma （MM） is a neoplasm of a terminally differentiated B-cell. Human myeloma cell lines were shown to be suitable model systems for use in various fields of the biological sciences. This study was aimed to investigate the genetic aberrations in human multiple myeloma cell lines. Interphase fluorescence in situ hybridization （FISH） with probes for the regions containing 13q14（RB-1）,13q14.3 （D13S19）,14q32 （IGHC/IGHV） ,1q12（CEP1）,17p13（TP53） were was used to delect 7 HMCL and 85 cases of newly diagnosed MM. FISH with LSI IGH/CCND1 ,LSI IGH/FGFR3 and LSI IGH/MAF probes were used to detect t（11;14）（q13;q32） ,t（4;14）（p16;q32） and t（14;16）（q32;q23） in HMCL and MM with 14q32 rearrangement. The results showed that molecular cytogenetic aberrations were found in all 7 HMCL,six （85.7%） HMCL simultaneously had 13q14,13q14.3 deletion. Chromosome 1q21 abnormality was found in six （33.3%） HMCL with at least 3 copies amplifications. Illegitimate 14q32 rearrangement was found in five （71.4%） HMCL,including one with t（11;14）,two with t（4;14） and three with t（14;16）. 17p13 deletion was detected in 5 HMCL. Chromosomal changes were observed in 85.9% of the 85 cases of newly diagnosed MM. The del（13）,1q12 amplification,del（17p）,14q32 rearrangement,t（11;14）,t（4;14）,t（14;16） were present in 44.7%,52.9%,20%,62.4%,27.1%,24.7% and 3.5% of the patients respectively. There was no significant difference in the prevalence of genetic abnormalities of del（13q）,14q32 rearrangement,1q12 amplification,t（11;14）,t（4;14） except del（17p） and t（14;16）. It is concluded that HMCL representative of the most aggressive phase of plasma cell neoplasms accumulated a large amount of genetic gberrations. Loss of p53 are strikingly common in HMCL suggesting that the impairment of the P53 tumor suppressor pathway is an important contributor to extramedullary tumor expansion.

关 键 词：骨髓瘤细胞系 遗传学异常 荧光原位杂交 

分 类 号：R733.3[医药卫生—肿瘤]