Different cytogenetic patterns in skeletal breast cancer metastases

Short-term cultures of breast cancer metastases to bone from two patients were analyzed cytogenetically. One metastasis had a complex hypotriploid karyotype with numerous marker chromosomes, whereas the other had simple karyotypic changes in three unrelated clones, 46,XX, t(4;11)(p14;p13)/45,XX,-19/46,XX,del(3)(p13p23), suggesting that the metastasis had originated from a simultaneous invasion of multiple cells from the primary tumor. The metastasis with complex chromosomal aberrations developed quickly as part of a clinically aggressive disease, whereas that with simple changes developed more than 20 years after the initial breast cancer diagnosis. Our findings therefore indicate that the tumor karyotype may play a role in determining the clinical course in patients with breast cancer. Genes Chromosom Cancer 16:72–74 (1996). © 1996 Wiley-Liss, Inc.     

Inv(14)(q11q32) in one of four different clones in a case of angioimmunoblastic lymphadenopathy

A case of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) with four cytogenetically different cell clones (49,XX,+5,+19,+21/47,XX,+X/46,XX,inv (14)(q11q32)/45,X,-X) is reported. To our knowledge, this is the first case of AILD with an inv(14)(q11q32), thus probably involving the T-cell receptor alpha-chain gene. The cytogenetic findings are discussed with respect to the possible progression of AILD to malignant lymphoma.     

Simple numerical chromosome aberrations in well-differentiated malignant epithelial tumors

Cytogenetic analysis of four well-differentiated malignant epithelial tumors revealed primary clones with only numerical abnormalities. The karyotypes were 49,XX, +5, +5, +7, +7, -17/50,XX, +5, +5, +7, +7, -17, +r in an adenocarcinoma of the lung; 47,XX, +3/47,XX, +5/47,XX, +7 in a squamous cell carcinoma of the epiglottis; 47,XX, +5/48,XX, +5, +10 in a squamous cell carcinoma developing in an ovarian dermoid cyst; and 52,XX, +5, +7, +8, +14, +15, +21 in a seropapillary ovarian adenocarcinoma. Also, in previously published cases exclusively numerical aberrations were much more common in highly differentiated epithelial tumors (22/74) than in moderately to low-differentiated carcinomas (13/281). Our findings and the literature data thus agree with a developmental scheme in which numerical changes, possibly reflecting an early-onset genomic instability in the tumor cells, may precede massive structural anomalies in the gradual malignization of epithelial tumors.     

Monozygotic twins discordant for trisomy 21 and maternal 21q inheritance: a complex series of events

We report on a monochorionic/diamniotic twin pregnancy discordant for trisomy 21. Amniocentesis (at 13(5/7) weeks) was performed following ultrasound signs of hydrops and cystic hygroma in twin 1 (T1). Prenatal karyotype showed non-mosaic trisomy 21 in T1 (47,XX,+21[7]), and low-grade mosaic trisomy 21 in twin 2 (T2) (47,XX,+21[2]/46,XX[19]). Post mortem examination of fetal skin, kidneys and lungs confirmed trisomy 21 in T1 (47,XX,+21[548]) and the placenta (47,XX,+21[200]). T2 had a normal karyotype (46,XX[648]). Analysis of microsatellite polymorphisms in multiple samples from the placenta, hand, lungs, kidneys and the umbilical cords of both twins confirmed monozygosity for all loci tested, and trisomy 21 in T1. Unexpectedly, T1 and T2 inherited different maternal alleles for markers of the most distal 4 Mbp of 21q. At least four successive events are needed to explain the genetic status of both twins and include maternal MI premature chromatids separation or maternal II meiotic nondisjunction and post-zygotic events such as, chromosome rescue, nondisjunction, an/or recombination.     

Multiple apparently unrelated clonal chromosome abnormalities in a squamous cell carcinoma of the tongue

We have cytogenetically examined short-term cultures from a squamous cell carcinoma of the tongue, a tumor type in which chromosome aberrations hitherto have not been reported. No less than 12 pseudodiploid clones were detected, giving the tumor karyotype 46,X,der(X)t(X;1)(q26;p32),der(1)(Xqter→Xq26::1p32→cen→1q42:),del(13)(q11q21),t(15;?) (q26;?)/46,XX,t(1;?)(p34;?),inv(2)(p21q11)/46,XX,t(1;10)(p32;q24)/46,XX,+der(1)(12pter→ 12p11::1p11→cen→1q32::11q13→11q32→1q42:),del(11)(q13q22), - 12, der(17)t(1:17) (q42;p13)/46,XX,inv(1)(p22q44)/47,XX,del(1)(q32),der(17)t(1:17)(p22;q25),der(1)inv(1) (q25q44)t(1;17)(p22;q25),ins(14;7)(q11;q22q36), + 14/46,XX,t(1;4)(q23;q35)/46,XX,t(1;21) (q25;q22),t(2;10)(q31;q26),t(22;?)(q12;?)/46,XX,del(1)(q32)/46,XX,t(1;8)(q44;q21)/46,XX, t(2;21)(q11;p11)/46,XX,t(9;11)(q34;q13). The large number of apparently unrelated abnormalities leads us to suggest that the carcinoma may have been of multiclonal origin.     

Analysis of the steroidogenic acute regulatory protein (StAR) gene in Japanese patients with congenital lipoid adrenal hyperplasia

Genomic DNA from 19 Japanese patients with congenital lipoid adrenal hyperplasia (lipoid CAH) representing 16 different families was examined to identify the genetic alterations of steroidogenic acute regulatory protein (StAR). Ten of 19 patients had a 46,XX karyotype and nine had a 46,XY karyotype. Six of the 46,XX patients have experienced spontaneous pubertal changes including breast development and irregular menstruation whereas none of the 46,XY subjects displayed pubertal changes. Eight different mutations were identified. Sixteen patients were either homozygotes or compound heterozygotes for the Q258X mutation. The seven other mutations identified were 189delG, 246insG, 564del13bp, 838delA, Q212X, A218V and M225T. The 189delG, 246insG, 546del13bp and Q212X mutants encode truncated proteins. COS-1 cells transfected with expression vectors encoding cDNAs for the mutant StAR proteins which affect the C-terminus, 838delA, A218V and Q258X, exhibited no steroidogenesis enhancing activity. However, the M225T mutant retained some steroidogenic activity. The patient with the M225T mutation had late onset of this disorder and some capacity to secrete testosterone in response to hCG. These findings suggest: (i) that the Q258X mutation can be used as a genetic marker for the screening of Japanese for lipoid CAH, (ii) that the C-terminus of StAR plays an important role in the protein's activity and (iii) that there are differences in the extent of functional impairment of the testis and ovaries in lipoid CAH.      

Complex karyotypic anomalies, including an i(5p) marker chromosome, in malignant mixed mesodermal tumor of the ovary

Cytogenetic analysis of short-term cultures initiated from an ovarian malignant mixed mesodermal tumor yielded the following karyotype: 59-61, XX,t(1;?)(p36;?), +t(1;9) (q43;q21), +t(2;?)(p25;?), +i(5p), +i(5p), +7, +t(7;?)(p13;?), +8,der(11) (pter----cen----q23::q13----q23::q13----q23::?), +12, + der(13)t(13;15)(q21;q15), -15,der(16) (16qter----cen----16p13::hsr::8q21----8qter), +19, + der(20)t(X;20)(q13;p13), -22, +4 - 6mar. Because the only other cytogenetically characterized ovarian neoplasm of this rare histopathologic subtype also had a small metacentric marker interpreted as an isochromosome for the short arm of a B-group chromosome, we suggest that i(5p) constitutes a nonrandom anomaly in mixed mesodermal tumors.     

Reversibility of lymphokine-induced NK-like activity in virus-specific cytotoxic T-lymphocyte clones.

A limiting dilution microculture system, supplemented with a source of interleukin-2 (IL-2), was employed to evaluate the frequency of Moloney-murine leukaemia/sarcoma virus (M-MuLV/M-MSV)-specific cytotoxic T-lymphocyte precursors (CTL-p) which also exhibited NK-like activity. Spleen cells, obtained from M-MuLV/M-MSV regressor mice, were restimulated in bulk secondary mixed leucocyte-tumour cell cultures (MLTC), and subsequently plated in a culture medium supplemented with two different supernatants (SN) produced following PMA-stimulation of the same EL-4 thymoma cell line. SN 20, obtained from the cell line maintained in vitro, contained IL-2 and only negligible amounts (less than 3 U/ml) of interferon (IFN), while SN 19, obtained after passage of the ascitic form of EL-4 thymoma in syngeneic mice, contained both IL-2 and IFN in high titres. The frequency of CTL-p specific for MBL-2 lymphoma cells was high and comparable in cultures supplemented with both SN (1/2 X 84 cells and 1/2 X 40 cells, respectively), while the frequency of CTL-p directed against NK-susceptible YAC-1 target cells was low in SN 20 (1/90 cells) and high in SN 19 (1/5 X 40 cells). An analysis of individual microcultures established at low cell dose (1 cell/well) indicated that specific and NK-like activity could be ascribed to the same precursor cells. Furthermore, using different long-term CTL clones, we observed that, after passage in SN 20, double-reactive clones gradually lose the capacity to lyse NK-susceptible targets, while most of MBL-2 specific clones acquired NK-like activity following a few passages in SN 19. Therefore, the induction of NK-like activity is reversible and may be modulated by soluble factors present in supernatant in which CTL clones are maintained. Double-reactive clones were unable to lyse NK-resistant allogeneic tumour cells or normal syngeneic blast cells. A few clones cross-reacting with H-2d alloantigens also exhibited NK-like activity when maintained in SN 19. The different pattern of CTL clone activity was associated with a morphological change in the clones themselves: the acquisition of double activity was accompanied by an increase in cell size and the appearance of numerous cytoplasmic granules. All CTL clones were phenotypically Thy-1+ and Lyt-2+ on indirect immunofluorescence and complement-dependent cytotoxicity investigation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Multiple unrelated clonal chromosome abnormalities in an in situ squamous cell carcinoma of the skin

We have cytogenetically analyzed short-term cultures from an in situ squamous cell carcinoma of the skin (Bowen's disease). The following mosaic tumor karyotype was found: 46,XX, -1, +der(1)(pter----p22::q11----cen----p22:), -9, +der(9)t(1;9)(q11; p24)/46,XX,t(3;6) (q21;p21)/46,XX,t(5;14)(q13;q24),t(7;18)(q32;q11)/46,XX,t(8;11)(p22;q13) /46, XX,t(8;11) (p22;q13),t(15;17) (q13;q24)/46,XX,t(12;15)(q12;p11). None of the rearrangements correspond to previously known cancer-associated abnormalities. Two of the clones are obviously related, and it is reasonable to assume that the t(15;17) developed as an evolutionary change in a cell that already contained t(8;11)(p22;q13). Since five clones without cytogenetic similarities were found in this in situ skin carcinoma, we suggest that the tumor was of polyclonal origin. It is impossible to decide whether all, or indeed any, of the visible abnormalities constitute pathogenetically essential primary changes, or merely represent chromosomal markers of secondary importance in tumorigenesis.     

Cytogenetic characteristics of T-lymphoid cell strains ofPapio hamadryas

A detailed cytogenetic study of three T-lymphoid strains ofPapio hamadryas cells was carried out. Two strains, LNPH-5(T) and SPH-7(T), had normal karyotype 42,XX at the 43rd and 56th passages. After long culturing the initial SPH-7(T) cells were completely replaced by the 87th passage by cells with a pseudodiploid karyotype 42,XX,der(1), der(2), der(13), der(14), der(X). SPH-8(T) strain (36th passage) contained two cell clones, with predominant normal (42,XY) and pseudodiploid 42,XY, der(7)t(7;X) (qter;pter), del(13)(q21),del(14)(q16) karyotype. The patterns of karyotypic variability of simian and human T-lymphoid cell strains are similar. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 126, No. 10, pp. 444–447, October, 1998     

A boy with true hermaphroditism and sex chromosome mosaicism and a fertile woman with Turner mosaicism in a family with a translocation 8p:19p

In a family with a balanced translocation t(8;19)(p21p13), there was a boy with true hermaphroditism and a karyotype 46, XX/46XY, t(8p;19p), and a woman with Turner mosaicism 46, XX, t(8p;19p)/45, X, t(8p;19p). Both of them had whole body chimerism, which in the boy and possibly also in the woman was due to the occurrence of double fertilization followed by fusion of the zygotes. The pathogenetic importance of the translocation for the development of these aberrations, and the clinical picture in the two patients are discussed.     

Translocation 1;19--a new cytogenetic abnormality in acute lymphocytic leukemia

A study of the chromosomes of 125 consecutive patients with acute lymphocytic leukemia (ALL) showed the same translocation between chromosomes #1 and #19 in 5 patients. In 4 of the 5, the t(1;19)(q21;q13) was present at diagnosis. The fifth patient, who had Philadelphia chromosome positive (Ph1+) ALL, developed t(1;19) in first relapse. Trisomy 1q was involved in 2 of the 5 patients; 3 patients had additional abnormalities. All patients had low white cell counts at presentation (less than 35 X 10(9)/L), and the 4 patients tested had common ALL antigen (CALLA) positive leukemic blast cells. All achieved complete remission, including the Ph1+ ALL patient in first relapse, and survival times ranged from 4 to 21+ mo from the time the t(1;19) first appeared. Our data suggest that t(1;19) is a previously unrecognized nonrandom structural abnormality in ALL that is also found in other lymphoid malignancies. Unlike the other specific translocations, it is not associated with a poor prognosis.     

Cytogenetic analysis of ependymoma and teratoma of the ovary

Cytogenetic analysis was performed on G-banded chromosomes from short-term cultures established from surgically removed tumor material from two patients. A karyotype of 56, XX, +5, +7, +7, +8, +13, +13, +18, +19, +20, +21 was found in one ovarian ependymoma. The grade 3 immature teratoma showed a karyotype of 47,XX, +3.     

Cytogenetic studies of eight primary gastric cancers.

Cytogenetic analysis was performed on eight primary gastric cancers. Three of them had simple chromosome changes: 47,XX,+X/48,XX,+X,+X; 48,XX,+8,+19,t(3;5) (q21;q31) and 47,XY,+del(7)(q22). The five others had complicated chromosome changes; both 3p- and 7q- were noted in four cases and i(5p) was noted in two cases.     

T-cell receptor beta gene rearrangements in clones derived from human CD4-8- cells expressing natural killer cell activity.

Clones derived from highly purified human peripheral blood Leu 19+ cells in the presence of phytohaemagglutinin (PHA) and interleukin-2 (IL-2) expressed cytotoxic activity against natural killer (NK)-resistant as well as NK-sensitive targets. All 66 clones analysed had a germ line configuration of T-cell receptor (TCR) beta genes and 38/40 also had unrearranged TCR gamma genes. The two exceptions were both CD3+ clones, but these did not have a cytotoxic repertoire noticeably different from CD3- clones without TCR gamma gene rearrangements. Clones were also obtained from highly purified CD4-8- cells, most of which were also cytotoxic for NK-resistant and NK-sensitive targets. About 90% of these clones were CD3+ but only around 50% remained negative for CD4 and CD8 while a significant number (12.7%) were positive for both CD4 and CD8. All clones analysed had rearranged TCR gamma genes and most had also rearranged TCR beta genes, including 20/25 of the clones which were CD3+4-8-. Many of the clones showed two rearrangements of TCR beta genes, and 3/4 CD3- clones had rearranged TCR beta as well as TCR gamma genes. There was no correlation between cytotoxic activity and TCR gene status or phenotype of these CD4-8- derived clones, except that clones which were Leu 19+ tended to have higher cytotoxic activity against NK-sensitive and NK-resistant targets than Leu 19-clones. The results strongly indicate that TCR beta and gamma gene products are not involved in the cytotoxicity mediated by these clones. They also suggest that some CD4-8- cells may be capable of limited differentiation in vitro.     

Clonal chromosome abnormalities in two chemodectomas

Short-term cultures from two histologically benign chemodectomas, one from the carotid body and one from the vagal nerve, were analyzed cytogenetically. The former had a small abnormal clone with the karyotype 46,XX,t(3;19)(q21;q13),t(12;15) (p13;q12–14), whereas the majority of the cells from the latter tumor displayed two related abnormal clones: 46,XY, i(l)(q10)/46, idem,add(2)(q37). The findings add to the evidence that chemodectomas are heterogeneous neoplasms and suggest that the heterogeneity may possibly be associated with the site of origin. Genes Chromosom Cancer 15:178–181 (1996). © 1996 Wiley-Liss, Inc.     

Origin and mechanism of formation of 45,X/47,XX,+21 mosaicism in a fetus

Chromosome analysis of amniotic fluid cells from a 17-week-old fetus with a nuchal cystic hygroma showed a 45,X/47,XX,+21 karyotype. Analyses of cord blood lymphocytes, skin fibroblasts, amniotic membrane, and chorionic villi demonstrated both cell lines in various proportions. We studied the origin and mechanism of formation of the double mosaic aneuploid using Q-banded chromosomal heteromorphisms, and one RFLP, two VNTRs, one tetranucleotide repeat, 28 CA repeat markers, mapped to every member of chromosomes. The heteromorphic markers examined showed no discordant patterns in parent-to-child transmission or between the two cell lines except for those in chromosomes 21 and X. Fetal DNA was extracted from its established monoclonal fibroblast cell lines with 45,X or 47,XX,+21 karyotypes. Genotyping with the DNA markers showed that each cell line was identical at every locus, except for chromosome 21 or X loci, indicating that the fetus was not a chimera but a mosaic. The 21-trisomic cells had one paternal allele and two maternal heterozygous alleles at the D21S270 locus, and the 45,X (21-disomic) cells had two biparental alleles. Alleles at two X chromosomal loci, DXS991 and DXS8057, were biparental in the 47,XX,+21 cells, whereas only the paternal allele was retained in the 45,X cells. Based on these findings, we concluded that the fetus started as a 47,XX,+21 zygote that had resulted from nondisjunction at the maternal first meiotic division and that one each of the maternally derived chromosomes 21 and X was lost during an early mitotic division, leading to the mosaicism. Am. J. Med. Genet. 75:432-437, 1998. © 1998 Wiley-Liss, Inc.     

A variant Burkitt-type translocation (2p-;8q+) in a patient with diffuse large cell lymphoma

A case of diffuse large cell lymphoma with t(2p-;8q+) is reported. Immunologically the lymphoma cells were shown to be of B-cell origin and positive for surface gamma and kappa chains, B4, CALLA, and Ia1 markers. Karyotypically three major clones were detected: 47,XX, + 12,t(2;8)(p11-13;q24) (52%); 47,XX, + 12 (26%); and 46,XX,t(2;8)(p11-13;q24) (15%). A t(2p-;8q +) has been exclusively reported in cases of Burkitt's lymphoma or Burkitt-type acute lymphocytic leukemia. The present case is the first one with t(2p-;8q +) observed in non-Burkitt-type lymphoid malignancy of the B-cell lineage. The t(2p-;8q +) may play a primary role in the early stage of transformation of B cells, and trisomy 12 may provide them secondarily with an advantage for tumor progression. The phenotypic pictures provided by 8q24 rearrangements seem to be heterogeneous, as previously suggested.     

Cytogenetic and fluorescence in situ hybridization analysis of breast fibroadenomas.

We report cytogenetic and fluorescence in situ hybridization (FISH) analysis findings in 7 patients with breast fibroadenomas (FA). Three patients were cytogenetically abnormal. One patient had a translocation t(3;5)(p22;q13), the second had trisomy 8, and the third two clones, 47, XX, +11 and 47,XX, +10.     

Chromosome aberrations in 35 primary ovarian carcinomas.

Cytogenetic analysis was performed on short-term cultures of primary ovarian carcinomas from 62 patients. Cytogenetic analysis was successful in 59 cases. Clonal chromosome aberrations were detected in 35 tumors. Only numerical changes or a single structural change were found in five carcinomas: trisomy 12 was the sole anomaly in two tumors, one tumor had the karyotype 50,XX, + 5, + 7, + 12, + 14, a fourth tumor had a balanced t(1;5), and the fifth tumor had an unbalanced t(8;15). The fact that four of these five carcinomas were well differentiated suggests that simple karyotypic changes are generally characteristic of these less aggressive ovarian tumors. The majority of the cytogenetically abnormal tumors (n = 30) had complex karyotypes, with both numerical and structural aberrations and often hypodiploid or near-triploid stemlines. The numerical imbalances (comparison with the nearest euploid number) were mostly losses, in order of decreasing frequency -17, -22, -13, -8, -X, and -14. The structural aberrations were mostly deletions and unbalanced translocations. Recurrent loss of genetic material affected chromosome arms 1p, 3p, 6q, and 11p. The breakpoints of the clonal structural abnormalities clustered to several chromosome bands and segments: 19p13, 11p13-15, 1q21-23, 1p36, 19q13, 3p12-13, and 6q21-23. The most consistent change (16 tumors) was a 19p + marker, and in 12 of the tumors the 19p + markers looked alike.