Clinical and cytogenetic correlations of abnormal megakaryocytopoiesis in patients with acute leukemia and chronic myelogenous leukemia in blast crisis

It has been suggested that abnormalities of chromosome 3 at bands q21 and q26 are associated with the presence of increased numbers of abnormal megakaryocytes in patients with hematologic malignancies. The pretreatment bone marrows of 287 patients with leukemia (acute myeloid leukemia (AML), 225 patients; acute lymphocytic leukemia (ALL), 36 patients; or chronic myelogenous leukemia in blast crisis (CML-B), 26 patients) were reviewed to identify those with normal or increased numbers of megakaryocytes. Thirty-two patients with AML, one with ALL, and 10 with CML-B had normal or increased numbers of megakaryocytes. Of the 32 patients with AML, 19 patients had significant numbers of mononuclear or binuclear small megakaryocytes as well as megakaryocytes with separated nuclei ("micromegakaryocytes"). Cytogenetic analyses were obtained in 29 of 32 patients with AML and showed inv(3)(q21q26) (one patient); Ph1 (two patients); -5 and/or -7 (seven patients); normal karyotypes (10 patients). No patient with micromegakaryocytes had a chromosomal abnormality associated with a favorable prognosis. Overall, among 225 patients with AML, four had inv(3)(q21q26) or t(3;3)(q21;q26). Only one of these four patients had normal or increased numbers of megakaryocytes, although all four had micromegakaryocytes. One patient with CML-B had inv(3)(q21q26) but had decreased numbers of megakaryocytes and a platelet count of 24 x 10(3)/microliters. All five patients with abnormal chromosome 3 at bands q21 and q26 had additional cytogenetic abnormalities (Ph1 in two patients; -7 in three patients). Mean and median platelet counts were greater than 100,000/microliters for patients with marrow megakaryocytosis regardless of morphology, as well as for the patients with abnormalities involving 3q21 and 3q26. Abnormalities of megakaryocyte morphology, increases in the numbers of megakaryocytes, and normal to increased platelet counts are not uncommon in patients with acute leukemia and CML-B, and are not uniquely associated with changes involving chromosome 3.     

Acute minimally differentiated myeloid leukemia (M0) with inv(3)(q21q26)

We describe a 55-year-old Japanese man with acute minimally differentiated myeloid leukemia (M0) with an inversion in the long arm of chromosome 3, i.e., inv(3)(q21q26). Patients with this chromosomal abnormality usually show normal or elevated platelet counts. However, our case had a low platelet count with megakaryocytic dysplasia at diagnosis. Furthermore, the 3q21q26 aberration is generally detected in patients with acute myelogenous leukemia (AML) or myelodysplastic syndrome. To the best of our knowledge, it has also been reported in two cases of AML-M0 with 3q21q26 and this is the third case of AML-M0 with 3q21q26. Thus it is suggested that there is some relationship between this type of karyotype abnormality and leukemogenesis and/or thrombopoiesis.     

A novel translocation t(3;22)(q21;q11) involving 3q21 in myelodysplastic syndrome-derived overt leukemia with thrombocytosis

We report here a novel translocation t(3;22)(q21;q11) in myelodysplastic syndrome (MDS)-derived overt leukemia with thrombocytosis. A 44-year-old female was initially diagnosed as MDS with a low platelet count and normal karyotype. After 4 months, blood leukemic cells and platelets rapidly increased concomitantly and a diagnosis of acute myeloblastic leukemia (AML M1) was made. Chromosome analysis showed 46, XX, t(3;22)(q21;q11) in 14 of 20 metaphases. Fluorescence in situ hybridization analysis confirmed both the der(3)t(3;22) and the der(22)t(3;22). Our results suggest that unidentified gene(s) at 3q21 breakpoint may be implicated in the pathogenesis of abnormal thrombopoiesis as observed in the 3q21q26 syndrome.     

Clinical correlations of the 3q21;q26 cytogenetic anomaly. A leukemic or myelodysplastic syndrome with preserved or increased platelet production and lack of response to cytotoxic drug therapy

Three patients presenting with acute leukemic disorder and chromosome 3 rearrangement involving bands q21;q26 are reported, and the literature on chromosome 3q abnormalities is reviewed. All reported patients carrying a paracentric 3q inversion or a translocation 3;3 with breakpoints in q21;q26 had a myelodysplastic or acute leukemic disorder with a normal or elevated platelet count and lack of response to cytotoxic drug therapy. They showed an associated incidence of -7 or 7q- anomalies higher than de novo acute leukemia and appear to constitute a definite subgroup of the leukemic disorders with very poor prognosis. The majority of patients showing other chromosome 3 long arm rearrangements showed evidence of leukemic process, were in blastic crisis, or had been exposed to chemotherapy, exhibiting also a higher incidence of associated -5 or -7 cytogenetic abnormalities than is observed in patients not exposed to toxic agents.     

肺癌患者血小板和巨核细胞观察

本文对３０例肺癌患者血象和骨髓象进行了观察，发现有６．７％的患者血小板减少，２３．３％血小板增多，且血小板数和骨髓巨核细胞数无相关性；并在骨髓象中发现巨核细胞的形态学改变，出现小巨核细胞、核分叶过多巨核细胞、巨大巨核细胞和吞噬血细胞的巨核细胞等病理性改变。     

Cytogenetic Abnormalities in B-Immunoblastic Lymphoma

We have considered the cytogenetic abnormalities present in 27 unpublished cases of B-immunoblastic lymphoma. Among these 27 patients, the chromosome changes were heterogeneous and complex. The chromosomes most commonly gained were 3 (44% of cases), 18 (44%), 6 (30%) and 11 (30%). The most common structural abnormalities involved band 14q32 (26%), band 18q21 (15%) and bands 6q 16-21 (19%). Study of these 27 immunoblastic lymphomas did not allow us to tentatively identify a common primary cytogenetic abnormality unique to B-immunoblastic lymphoma, however, a translocation at 14q32 may be the primary cytogenetic lesion in some of the cases. Rather, we have added to the number of abnormalities reported in immunoblastic lymphoma and in non-Hodgkin's lymphoma in general.     

Stimulation of CFU-Mk colony growth by normal plasma and plasma from myelodysplastic patients

The ability of plasma from myelodysplastic patients to support the clonal growth of normal megakaryocyte progenitors (CFU-Mk) was compared with that of plasma from normal subjects. The resultant megakaryocyte colonies were expressed as a plasma factor index megakaryocyte (PFI-Mk). All cultures included PHA-LCM and medium conditioned by the human bladder carcinoma cell line 5637, and some of them had EPO. PFI-Mk (MDS) was significantly lower than PFI-Mk (normal), both with and without EPO. A positive correlation was found between megakaryocyte and platelet count in normal subjects, but was not present in MDS patients. There was no correlation between platelet count and PFI-Mk in neither group. In MDS there was a negative correlation between megakaryocyte number and PFI-Mk, both with and without EPO. Although, the mean megakaryocyte number in MDS and in normal bone marrow was similar, the proportion of immature megakaryocytes was much higher in MDS. Previous work indicates an abnormal clonal origin of megakaryocytes in MDS. The present study suggests that abnormal plasma factors affects megakaryocytopoiesis in this condition.     

Acute Leukemia with Structural Rearrangements of Chromosome 3

Cytogenetic investigations have distinguished at least 3 distinct clinical-cytogenetic syndromes of hematopoietic malignancy with structural rearrangement of 3q. The majority of cases have breakpoints at both 3q21 and 3q26, frequently associated with monosomy 7, abnormal thrombopoiesis, and adverse outcome. Cases with only one of these breakpoints may have milder features of the syndrome. A subgroup with t(3q;5q) occurs in younger patients, occasionally with megakaryocytic dysplasia but rarely having thrombocytosis. The t(3;21) is encountered in secondary leukemias or after chemotherapy of myeloproliferative disorders. The genetic deregulations associated with each of these syndromes involve distinct genes on 3q. The majority of cases of acute leukemias with 3q rearrangements have a poor prognosis and do not respond to current modes of therapy.     

Specific abnormalities of chromosome 14 in patients with acute type of adult T-cell leukemia/lymphoma

Chromosome analysis was performed on 1 patient with diffuse lymphoma of mixed type by histologic diagnosis and on 7 patients with the acute type of adult T-cell leukemia (ATL). Specific abnormalities in chromosome 14 at break band q11 with the assigned locus of the alpha-chain gene of the T-cell antigen receptor were identified in 6 of 8 patients. Inv(14) (q11q32) was found in 2 patients and translocation of chromosome 14 at break band q11 was observed in 4. Donor chromosomes involved in translocation of the 14q11 varied, i.e., chromosomes 3, 7 or X, with the exception of one patient whose donor chromosome origin could not be determined. The breakpoint in chromosome 3 was in band p25, a region reported to include the locus of the c-raf-I oncogene. In chromosome 7, it was in band p11, a region reported to include the locus of the c-erb-B oncogene, and in the sex chromosome X, it was in band q11. One patient also had a chromosome 14 aberration at break band q32. Of the 2 remaining patients, one had lost chromosome 14 and the other had an isochromosome 14q. Our observation and other reported findings suggest that the rearrangement of chromosome 14 at break band q11 is specific for lymphoma-type or acute-type ATL patients, and aberrations of proto-oncogene expression or the coding sequence by recombination involving a T-cell antigen receptor gene due to chromosome inversion or chromosome translocation may play an important role in T-cell neoplasia including ATL.     

Recurrent chromosome changes at 3p21 in Epstein-Barr virus-transformed cells derived from human prolymphocytic leukemia

Cytogenetic studies of Epstein-Barr virus-transformed lymphoblastoid cells obtained from a patient with prolymphocytic leukemia revealed the cells to contain recurrent chromosome aberrations involving band 3p21. The in vivo leukemic cells from which the cell lines originated contained a der(3)t(3;17?)(p21;q11?) and a der(13)t(13;3)(q34;p21), as well as numerical (-Y, -8, and -17) and other structural [8p- and der(8)t(8;?)(q13?;?)] changes. The cells in established culture showed additional chromosome aberrations involving band 3p21 of the previously normal and abnormal chromosomes 3. After 200 days of culture, the cells contained a new translocation, i.e., t(3;14)(p21;q32). The cells showed further chromosomal breakage and reunion at band 3p21 at later passages. The affected chromosomal band (3p21) is close to one of the constitutive fragile sites, i.e., 3p14.2. Northern blot analysis of the messenger RNA of the cultured cells did not show an increased or altered expression of the c-raf-1 protooncogene (located at 3p25) when compared with the mRNA of Epstein-Barr virus-transformed lymphoblastoid cell lines from normal subjects with a normal karyotype. Also, the established cells did not show DNA rearrangements or RNA alterations of the c-myc gene.     

A newly established human lymphoma cell line, FL-18, carrying a 14;18 translocation

A new human cell line named FL-18, carrying a 14;18 translocation [t(14;18)(q32;q21)], was established from a Japanese patient with follicular small cleaved cell lymphoma. The established FL-18 cells had monoclonal surface immunoglobulins (IgG-kappa) and were negative for Epstein-Barr virus nuclear antigens. The high-resolution banding technique indicated the breakpoints of chromosomes 14 and 18 involved in the characteristic 14;18 translocation to be at sub-bands 14q32.3 and 18q21.3, respectively. The FL-18 cell line should be useful for studying oncogenic events associated with 14;18 translocation and karyotype evolution.     

Effect of friend leukemia virus on megakaryocytes and platelets in mice

The thrombocytopenia that follows infection of C3H/Bi mice with Friend leukemia virus (FV) has been investigated. At various times after infection, megakaryocytes in bone marrow and spleen were collected on Nuclepore filters, stained with the Feulgen procedure, and counted. At the same times, platelets in circulating blood were enumerated by the direct method of Brecher et al., 1963 We found that megakaryocyte numbers decreased within 3 days of infection; platelet numbers began to fall at 7 to 9 days. At 11 days megakaryocyte numbers in femoral marrow and spleen were reduced to less than one-third of control values, platelets to about one-quarter of control values. The data are compatible with the hypothesis that the thrombocytopenia which occurs after FV infection in mice results from the reduction in megakaryocytes that is brought about by the infection. Whether all classes of megakaryocytes are equally affected by FV was next examined. At various times following infection, megakaryocytes in marrow suspensions were concentrated by unit gravity sedimentation and microspectrophotometric measurements of nuclear DNA content were made. The kinetic data showed that, following FV infection, megakaryocytes of high nuclear DNA content were markedly reduced in number, but those of low nuclear DNA content were relatively unaffected. Thus the virus infection may interfere with the normal sequential doubling in DNA content of megakaryocytes and/or it may lead to preferential elimination by the host of those megakaryocytes in the higher DNA classes.     

采用对比基因杂交技术建立嗅神经母细胞瘤基因畸变模型

目的建立嗅神经母细胞瘤(ENB)的基因畸变模型并分析其特征。方法采用对比基因杂交(CGH)技术对12例原发、3例复发、7例转移ENB标本进行基因检测,用特殊软件系统对所采集照片的荧光信号进行量化分析,确定肿瘤DNA与正常对照DNA之间的基因差别。结果ENB常见DNA丢失主要见于1P、2q、3p／q、4p／q、5p／q、6q、8p／q、9p、10p／q、11P、12q、13q、18q和21q,DNA过度表达见于1p、7q、9q、1lq、14q、16p／q、17p／q、19p／q、20p／q和22p／q。ENB的1p21-p31DNA丢失与该类肿瘤患者预后差有明显相关性。死于ENB的患者均具有以下共性：1p21-p31DNA丢失、临床分期为C或D期、分化程度低(Ⅲ或Ⅳ级)。对4例发生转移、复发的患者,将检测结果与其原发病灶基因畸变情况进行对比分析表明,转移、复发病灶与其原发病灶间具有高度的克隆一致性。结论采用CGH能建立ENB的典型基因畸变模型,该模型有助于解释此肿瘤的生物学特性并对其预后进行评估,并与神经母细胞瘤、小细胞肺癌以及头颈部鳞癌进行鉴别。     

t(3;5)(q21;q31) in a myelodysplastic syndrome

Cytogenetic analysis from the bone marrow of a patient with a myelodysplastic syndrome revealed the balanced translocation t(3;5)(q21;q31). Although this translocation has recently been described in six cases of AML, this is the first such observation in a preleukaemic syndrome. Subsequent evolution into RAEB and AML(M2) was noted without the acquisition of additional cytogenetic changes and complete remission achieved with conventional cytotoxic chemotherapy. The relationships with other acquired abnormalities of chromosomes 3 and 5 in MDS/AML are discussed.     

Two Cases of Translocation t(3;6)(p14;p22): A Non Random Chromosomal Abnormality?

It is generally accepted that the short arm of chromosome 6 is a likely site to be involved in chromosomal rearrangements of MDS/ANLL following radio/chemotherapy. We report here two cases of t(3;6)(p14;p22). One patient is a 55 years old male with a previous history of occupational exposure who developed, an acute megakaryoblastic leukemia after a preleukemic phase. Chromosome analysis showed a t(3;6)(p14;p22), associated with del (5)(q14q31), -7, with variations and a trend to hypoploidy. The second patient is a 33 years old man, with chronic myeloid leukemia treated with Hydroxyurea (HU), HU + $aL-IFN and $aL-IFN alone. The first cytogenetic study before treatment, showed a t(9;22)(q34;q11). In the following months the patient had simultaneously t(9;22)(q34;q11) + t(3;6)(p14;p22) in a minority and thereafter in all the mitoses, with progressive deterioration, megakaryocyte abnormalities, but no blast crisis. Our patients are compared with the only 5 other published cases with t(3;6)(p14;p22), who shared some common features, namely a past history of chemo/radiotherapy or exposure to chemical mutagens and an association with other, so-called “secondary” chromosome aberrations, on segments 3p, 5q, 7q, 12p and 17p. We suggest that this uncommon translocation t(3;6) is nonrandom. It is worth noting that band 6p21 is the site of pim 1 oncogen, and that a fragile site is located on band 3p14.     

Cytogenetic characterisation of Warthin's tumour

Warthin's tumour is a peculiar subtype of monomorphic adenomas of the salivary glands, frequently cystic, and that characteristically associates an epithelial glandular cell component to a dense lymphoid infiltrate. Short-term cultures from 12 Warthin's tumours of salivary glands, including 5 previously reported cases were successfully karyotyped and clonal numerical and/or structural changes were detected in 7 of them (58%). 3 cases showed numerical abnormalities with loss of chromosomes Y (2 cases) and X (1 case). The remaining 4 abnormal cases presented the following structural changes: complex translocation t(11;19;16)(q21;p12;p13.3); reciprocal translocations t(6;8)(p23;q22) and t(6;15)(p21;q15) (2 cases); and 1p22, 3p26, 11p13 changes. In 1 case, clonal numerical deviations (+7 and -Y) were concurrent with the structural rearrangement t(6;8). Two of these aberrations are suggested to be Warthin's tumour-associated: 11q;19p translocation has already been described in 3 cases, and structural rearrangements of 6p23 have also been reported in another case. Our study extends the cytogenetic information about Warthin's tumour and identifies two recurrent abnormalities —6p rearrangements and t(11;19) — specific for this salivary neoplasm.