【摘要】 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。更多还原

【Abstract】 Objective Using immunohistochemistry ( Immunohistochemistry,IHC) combined with fluorescence in situ hybridization ( fluorescence in situ hybridization,FISH) technology to detect multiple myeloma ( MM) common molecular cytogenetic abnormalities. Methods 20 cases of newly diagnosed MM patients diagnosed according to WHO criteria,take the bone marrow tissue embedded in paraffin and sliced; Application of IHC to FISH detection of CD138 + cells in bone marrow biopsies. Detected by 1q21 / RB1,D13S319 / p53,IGH three groups of sequence-specific gene probe,and 10 cases of non-malignant hematologic system disease in patients with bone marrow tissue sections as controls to establish probes for the detection threshold,the test results greater than the threshold positive,less than the threshold value was negative. Results ( 1) 16 cases detected in the molecular cytogenetic abnormalities in IHC combined with FISH technique detected in 20 cases of newly diagnosed MM patients ( 80. 0%) ,including 1q21 amplification in 5 cases ( 25. 0%) ,the RB1 deletion of 6 cases ( 30. 0%) ,D13S319 deletion 9 cases ( 45. 0% ) of p53 missing 3 cases ( 15. 0% ) ,the IGH gene rearrangement in 10 cases ( 50. 0% ) . Detection of abnormal 5 cases ( 25. 0% ) ,while there were two kinds of abnormalities in 6 cases ( 30. 0% ) ,three kinds of abnormalities in 4 cases ( 20. 0% ) ,four kinds of abnormal cases ( 5. 0% ) . ( 2) The IHC combined with FISH technique detection rate of 80% ,and chromosome G-banding technique detection rate of 10. 0% ,two sets of rate differences were significant ( P < 0. 05) . Prompt IHC combined with FISH detected MM molecular cytogenetic abnormalities was significantly superior to chromosome G-banding technique. ( 3) By age < 50 years old,50-60 years old,60-year-old grouping,molecular cytogenetic abnormalities detected in 5 cases ( 83. 3% ) ,respectively,in 6 cases ( 100%) ,5 cases ( 62. 5%) . Three groups of ages molecular cytogenetic abnormal results of the chi-square test age > 60 years compared with the other two groups there was significant difference ( P < 0. 05 ) . ( 4 ) Between chromosomal with genetic abnormalities of MM patients and with their clinical genotyping staging,that between p53 and IgG type with stage Ⅲa were significant( P < 0. 05) ,mainly IgG type and stage Ⅲ. Conclusions IHC combined with FISH to detect MM molecular cytogenetic abnormalities contribute to improve the detection efficiency. Between chromosomal with genetic abnormalities of MM patients and their clinical genotyping,staging,ages were relevant.更多还原