论文部分内容阅读
目的应用免疫组织化学(immunohistochemistry,IHC)联合荧光原位杂交(fluorescence in situhybridization,FISH)技术检测多发性骨髓瘤(multiple myeloma,MM)常见分子细胞遗传学异常。方法对按照WHO诊断标准确诊的20例初诊MM患者,取骨髓组织石蜡包埋并切片,应用IHC技术对骨髓石蜡切片进行CD138单克隆抗体标记,选取CD138+细胞丰富的骨髓石蜡切片,采用1q21/RB1、D13S319/p53、IGH三组序列特异性基因探针进行FISH检测。同时以10例非恶性血液病患者骨髓组织切片为对照建立各探针FISH检测的正常阈值,检测结果大于阈值为阳性,小于阈值为阴性。结果 (1)20例初诊MM患者中16例检出分子细胞遗传学异常(占80.0%),其中1q21扩增5例(占25.0%),RB1缺失6例(占30.0%),D13S319缺失9例(占45.0%),p53缺失3例(占15.0%),IGH基因重排10例(占50.0%)。检出1种异常者5例(占25.0%),同时有2种异常者6例(占30.0%),3种异常者4例(20.0%),4种异常者1例(占5.0%)。(2)IHC联合FISH技术检出率80%,而染色体G显带技术检出率10.0%,两组差异有统计学意义(P<0.05)。(3)按年龄<50岁、50～60岁、>60岁分组,分子细胞遗传学异常检出分别为5例(83.3%)、6例(100%)、5例(62.5%),经Fisher检验,年龄>60岁与其他两组比较差异有统计学意义(P<0.05)。(4)MM患者染色体与基因异常和其临床分型、分期之间,即p53基因与IgG型及Ⅲa期之间有关(P<0.05),染色体与基因异常以Ⅲ期和IgG型为主。结论 IHC联合FISH技术检测MM分子细胞遗传学异常有助于提高检测效率,明显优越于染色体G显带技术,同时可发现MM的分子细胞遗传学改变多数为复杂核型,且多数存在数目与结构异常。MM患者染色体和基因异常与其临床分型、分期、患者年龄之间具有相关性。
Objective To detect common molecular cytogenetic abnormalities in multiple myeloma (MM) by immunohistochemistry (IHC) combined with fluorescence in situ hybridization (FISH). Methods Twenty cases of newly diagnosed MM patients diagnosed according to the WHO diagnostic criteria were recruited. The bone marrow tissues were embedded in paraffin and sliced. The bone marrow paraffin sections were labeled with CD138 monoclonal antibody by IHC. The bone marrow paraffin sections enriched in CD138 + cells were selected. , D13S319 / p53, IGH three groups of sequence-specific gene probes FISH detection. At the same time, 10 normal bone marrow sections of non-hematologic malignancies were used as control to establish the normal threshold value of FISH for each probe. The detection results were more than the threshold value and less than the threshold value. Results (1) The molecular cytogenetic abnormalities (80.0%) were detected in 16 out of 20 newly diagnosed MM patients, of which 5 cases (25.0%) were amplified by 1q21, 6 cases (30.0%) were RB1 deletion, 9 (45.0%), p53 deletion in 3 cases (15.0%) and IGH gene rearrangement in 10 cases (50.0%). There were 5 abnormal cases (25.0%), 2 abnormalities in 6 cases (30.0%), 3 abnormalities in 4 cases (20.0%), 4 abnormalities in 1 case (5.0%), . (2) The detection rate of IHC combined with FISH was 80%, while the detection rate of chromosome G banding was 10.0%. There was significant difference between the two groups (P <0.05). (3) According to the age <50 years old, 50-60 years old,> 60 years old, the molecular cytogenetic abnormalities were detected in 5 cases (83.3%), 6 cases (100%) and 5 cases (62.5% Fisher test, age> 60 years old compared with the other two groups the difference was statistically significant (P <0.05). (4) There was a correlation between chromosomal and genetic abnormalities in MM patients and their clinical stage and staging, that is, between p53 gene and IgG and Ⅲa (P <0.05). Chromosome and gene abnormalities were mainly in stage Ⅲ and IgG. Conclusion IHC combined with FISH detection of MM molecular cytogenetic abnormalities help to improve the detection efficiency was significantly superior to the chromosome G banding technology, and found that most of the molecular cytogenetic changes in MM complex karyotype, and most of the number and structure abnormal. MM patients with chromosomal and genetic abnormalities and their clinical classification, staging, patient age has a correlation between.