中华临床医师杂志(电子版)
中華臨床醫師雜誌(電子版)
중화림상의사잡지(전자판)
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
8期
3345-3350
,共6页
杨璐璐%聂玉%刘欣%汪健%王晓秋%王志华%郑昌成%王兴兵%朱小玉%朱薇波%孙自敏%蔡晓燕%吴竞生
楊璐璐%聶玉%劉訢%汪健%王曉鞦%王誌華%鄭昌成%王興兵%硃小玉%硃薇波%孫自敏%蔡曉燕%吳競生
양로로%섭옥%류흔%왕건%왕효추%왕지화%정창성%왕흥병%주소옥%주미파%손자민%채효연%오경생
多发性骨髓瘤%免疫组织化学%原位杂交,荧光%细胞遗传学分析
多髮性骨髓瘤%免疫組織化學%原位雜交,熒光%細胞遺傳學分析
다발성골수류%면역조직화학%원위잡교,형광%세포유전학분석
Multiple myeloma%Immunohistochemistry%Situ hybridization,fluorescence%Molecular cytogenetic abnormalities
目的应用免疫组织化学( immunohistochemistry ,IHC)联合荧光原位杂交( fluorescence in situ hybridization,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。
目的應用免疫組織化學( immunohistochemistry ,IHC)聯閤熒光原位雜交( fluorescence in situ hybridization,FISH)技術檢測多髮性骨髓瘤(multiple myeloma,MM)常見分子細胞遺傳學異常。方法對按照WHO診斷標準確診的20例初診MM患者,取骨髓組織石蠟包埋併切片,應用IHC技術對骨髓石蠟切片進行CD138單剋隆抗體標記,選取CD138+細胞豐富的骨髓石蠟切片,採用1q21/RB1、D13S319/p53、IGH三組序列特異性基因探針進行FISH檢測。同時以10例非噁性血液病患者骨髓組織切片為對照建立各探針FISH檢測的正常閾值,檢測結果大于閾值為暘性,小于閾值為陰性。結果(1)20例初診MM患者中16例檢齣分子細胞遺傳學異常(佔80.0%),其中1q21擴增5例(佔25.0%),RB1缺失6例(佔30.0%),D13S319缺失9例(佔45.0%),p53缺失3例(佔15.0%),IGH基因重排10例(佔50.0%)。檢齣1種異常者5例(佔25.0%),同時有2種異常者6例(佔30.0%),3種異常者4例(20.0%),4種異常者1例(佔5.0%)。(2) IHC聯閤FISH技術檢齣率80%,而染色體G顯帶技術檢齣率10.0%,兩組差異有統計學意義( P<0.05)。(3)按年齡<50歲、50~60歲、>60歲分組,分子細胞遺傳學異常檢齣分彆為5例(83.3%)、6例(100%)、5例(62.5%),經Fisher檢驗,年齡>60歲與其他兩組比較差異有統計學意義(P<0.05)。(4)MM患者染色體與基因異常和其臨床分型、分期之間,即p53基因與IgG型及Ⅲa期之間有關( P<0.05),染色體與基因異常以Ⅲ期和IgG型為主。結論 IHC聯閤FISH技術檢測MM分子細胞遺傳學異常有助于提高檢測效率,明顯優越于染色體G顯帶技術,同時可髮現MM的分子細胞遺傳學改變多數為複雜覈型,且多數存在數目與結構異常。 MM患者染色體和基因異常與其臨床分型、分期、患者年齡之間具有相關性。
목적응용면역조직화학( immunohistochemistry ,IHC)연합형광원위잡교( fluorescence in situ hybridization,FISH)기술검측다발성골수류(multiple myeloma,MM)상견분자세포유전학이상。방법대안조WHO진단표준학진적20례초진MM환자,취골수조직석사포매병절편,응용IHC기술대골수석사절편진행CD138단극륭항체표기,선취CD138+세포봉부적골수석사절편,채용1q21/RB1、D13S319/p53、IGH삼조서렬특이성기인탐침진행FISH검측。동시이10례비악성혈액병환자골수조직절편위대조건립각탐침FISH검측적정상역치,검측결과대우역치위양성,소우역치위음성。결과(1)20례초진MM환자중16례검출분자세포유전학이상(점80.0%),기중1q21확증5례(점25.0%),RB1결실6례(점30.0%),D13S319결실9례(점45.0%),p53결실3례(점15.0%),IGH기인중배10례(점50.0%)。검출1충이상자5례(점25.0%),동시유2충이상자6례(점30.0%),3충이상자4례(20.0%),4충이상자1례(점5.0%)。(2) IHC연합FISH기술검출솔80%,이염색체G현대기술검출솔10.0%,량조차이유통계학의의( P<0.05)。(3)안년령<50세、50~60세、>60세분조,분자세포유전학이상검출분별위5례(83.3%)、6례(100%)、5례(62.5%),경Fisher검험,년령>60세여기타량조비교차이유통계학의의(P<0.05)。(4)MM환자염색체여기인이상화기림상분형、분기지간,즉p53기인여IgG형급Ⅲa기지간유관( P<0.05),염색체여기인이상이Ⅲ기화IgG형위주。결론 IHC연합FISH기술검측MM분자세포유전학이상유조우제고검측효솔,명현우월우염색체G현대기술,동시가발현MM적분자세포유전학개변다수위복잡핵형,차다수존재수목여결구이상。 MM환자염색체화기인이상여기림상분형、분기、환자년령지간구유상관성。
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 .