凋亡抑制蛋白Survivin在骨髓增殖性疾病中的表达及其意义

【作者】 蓝海峰；

【导师】 杨仁池；

【作者基本信息】 中国协和医科大学 ， 内科学， 2008， 博士

【摘要】 第一部分Survivin在骨髓增殖性疾病中的表达及其意义【目的】探讨Survivin在骨髓增殖性疾病的表达与临床相关性【方法】采用western blot方法检测21例ET、12例PV、12例CML和5例正常对照骨髓单个核细胞Survivin的表达,采用K562细胞系作为阳性对照。以β-actin作为内参计算Survivin表达水平的相对值。应用线性回归分析Survivin表达与临床特征的相关性。【结果】ET、PV和CML患者骨髓中分别有5例(23.8%)、8例(66.8%)和10例(83.3%)Survivin表达呈阳性,5例正常对照均为阴性。PV和CML组阳性率明显高于ET组(P=0.02和P=0.001)。各组总Survivin表达水平与外周血WBC、巨核细胞数呈显著正相关。分组分析显示,在CML中其水平与外周血WBC计数、外周血幼稚细胞比例均显著正相关,但ET、PV中无此相关性。PV组中Survivin表达与外周血Hb水平及PLT计数呈正相关,而CML中呈负相关。ET中Survivin表达与外周血WBC、Hb、PLT数、JAK2V617F以及骨髓幼稚细胞等均无显著相关性。【结论】Survivin在CML和ET、PV中的不同表达反映了它在这些疾病中的不同作用。在CML中,Survivin的异常表达可能发生于原始阶段细胞并与外周血幼稚比例呈正相关,而PV和ET中在相对成熟阶段起作用。PV患者的高Hb的维持与Survivin的高表达有关,提示Survivin在红系过度和异常增生中起作用。第二部分原发性血小板增多症的临床特点分析为了解我国原发性血小板增多症(ET)患者临床特点及疾病自然过程,对本院1980年5月至2006年12月间诊治的ET患者进行回顾性分析。438例患者中,男201例,女237例,中位发病年龄48岁。有出血者101例(23.1%),有栓塞者86例(19.6%),同时有出血和血栓者13例(3.0%),脾大150例,肝大60例。无症状者149例(34%),在诊治其他疾病时发现者145例(33.1%)。初诊时中位血小板数为1000×109/L。255例做骨髓活检,以大而多分叶的成熟巨核细胞增生为主,其中51例局部伴有网状纤维增生。114例患者检出90例JAK2V617F突变阳性,阳性率78.9%。180例做染色体核型检查,6例(3.3%)染色体异常。261例随访＞12月,中位随访时间60月(12～300月)。17例(6.5%)进展为骨髓纤维化(MF)。1例转为真性红细胞增多症(PV)。1例6年后转为PV,20年后转为MF。转为急性单核细胞白血病(M5)、骨髓增生异常综合征(MDS)、多发性骨髓瘤(MM)各1例。ET是以血栓和出血为主要症状的慢性骨髓增殖性疾病,无症状者比例高,易漏诊误珍。主要进展为MF,大多预后较好。第三部分180例原发性血小板增多症的细胞遗传学分析【目的】总结我国原发性血小板增多症(ET)患者的细胞遗传学特点及其临床意义【方法】对我院180例ET患者的细胞遗传学结果进行回顾性分析,并探讨特异性染色体畸变(Chromosomal Abnormalities,CA)的临床意义。【结果】1、细胞遗传学结果:应用常规染色体检查(Conventional Cytogenetics,CC)患者总体的克隆性CA检出率为3.3%;染色体数目畸变中最多见的是三体是6、11,其次为11、15、22;最多见的单体为21,其次为8、13、14、18、Y。染色体结构畸变中最常累及的长短臂是9q、11q、20q。常累及的断裂点是9q12-31、11q14、20q11～12、5q13-31。2、相关的临床特点分析:核型正常组和核型异常组的临床特征未见显著性差异,治疗有效率前者高于后者。【结论】我国ET患者的CA发生率较西方国家稍低,以染色体数目和染色体片段的增减为主,很少发生易位改变。克隆性染色体异常多为单一克隆性异常或伴有其他亚克隆异常核型。CA可累及所有的染色体,我国ET患者中+8的发生率明显低于西方国家,而累及9和21号染色体的几率较高。更多还原

【Abstract】 Objective To investigate the expression of Survivin and its clinical implication in CMPD.Methods The expression of Survivin protein in bone marrow mononuclear cells was measured by western blot in 21 ET,12 PV and 12 chronic myelogenous leukemia(CML) patients as well as five healthy donors.K562 cells were tested as a positive control.β-actin was used as an internal control.The relative levels of Survivin expression in each sample were calculated by normalizing to the levels ofβ-actin.The correlation between Survivin expression levels and blood cell counts was determined by drawing the best-fit linear curve with Regression Analysis.Results Survivin expression was observed in 23.3%(5 of 21) cases in ET, 66.8%(8 of 12) in PV and 83.3%(10 of 12) in CML,but undetected in the five healthy donors.The positive rate of Survivin expression in PV and CML was markedly higher than that in ET(P=0.02 and P=0.001) and healthy group. Survivin expression levels in all patients correlated with WBC count of peripheral blood and with megakaryocyte count of bone marrow.When analyzing 3 groups of CML,ET and PV separatively,we found that Survivin expression levels correlated with peripheral blood WBC count and with the percentage of peripheral blood blast cell in CML group,but not in ET and PV groups. However,Survivin expression levels in PV group was associated with the amount of Hb.In contrast,the levels of Survivinwere reverse to the amount of Hb in CML group.Moreover,the levels of Survivin in ET group has no significant relationship with WBC count,Hb,PLT count,JAK2V617F or the percentage of blast cell in bone marrow.Conclusions The different profile of Survivin expression in CML,PV and ET might reflects its different roles in these diseases.In CML,the aberrant expression of Survivin might happen earlier in blast cell and correlated with the percentage of blast cells.The correlation of Survivin and Hb in PV suggests a role of Survivin in Excessive and abnormal erythropoiesis. To summarize the clinical feature and natural course of essential thrombocy-themia (ET) in China,A retrospective analysis was conducted in ET patients treated in our hospital during May 1980 and December 2006.In this period,438 patients(201 males and 237 females with a median age 48 years) were diagnosed in our hospital, hemorrhage occurred in 101 cases(23.1%),thrombosis occurred in 86 cases(19.6%), and 13 cases(3.0%) had both hemorrhage and thrombosis.Splenomegaly occurred in 150 cases and hepatomegaly occurred in 60 cases.One hundred and forty-nine cases (34%) had no symptoms at diagnosis and 145 cases(33.1%) confirmed by routine blood tests due to other diseases.The median platelet count at diagnosis was 1,000×10~9 /L(533～3,740×10~9/L).Bone marrow biopsy was performed in 255 cases showing mainly increase of enlarged mature megakaryocytes with hyperlobulated nuclei and local proliferation of reticular fiber were revealed in 51 cases.JAK2V617F mutation was detected in 90(78.9%) of the 114 patients.Karyotype analysis was performed in 180 cases and 6 cases(3.3%) had clonal chromosomal aberrations.Two hundred and sixty-one patients were followed-up over 12 months with a median follow-up period of 60 months(range from 12 to 300 months).Seventeen cases(6.5%) evolved into marrow fibrosis(MF) and one case evolved into polycythemia vera(PV).One case evolved into PV after 6 years and MF after 20 years.Three cases developed acute monocyte leukemia(MS),myelodysplastic syndrome(MDS) and multiple myeloma (MM),respectively.ET is a chronic myeloproliferative disorder characterized predominately by thrombocytosis and hemorrhage.The percentage of asymptomatic cases is high.The main sequelae of ET is MF and most cases have good prognosis. Objective To summarize the cytogenetic characteristics and their clinical significances of the patients with essential thrombocythemia(ET) in China.Methods Cytogenetic findings of 180 patients with ET were retrospectively analysed and the clinical implications of specific chromosomal abnormalities(CA) were investigated.Result1.Cytogenetic findings:clonal CAs were detected in 6 of 180 patients(3.3%) by conventional cytogenetics(CC);In the numerical aberrations,the most common trisomies were 6,1 and the most common monosomies were 21,followed by 8, 13,14,18 and Y.In the structural aberrations,the most frequently involved arms were 9q,11q and 20q,and the most frequently involved breakpoints were 9q12～31,11q14,20q11～12 and 5q13～31.2.Clinical implications:there was no significant difference between the clinical characteristics of chromosomal abnormalities group and normal control,however the latter have a higher efficiency in therapy.ConclusionsThe occurency of CA in patients with ET from China is mildly lower than those from western country.The chromosomes numerical aberrations and deletant or addend of chromosomes arms are main abnormal in CA,but translocation occur seldom.Clonal CAs often display single clonal CA or company with other subclonal CAs.All chromosomes can be involved in CAs.The occurancy of +8 in patients of ET from China is obviously lower than those from western country,but chromosome 9 and 21 can be involved more frequency.更多还原