Application of chromosome microdissection probes for elucidation of BCR- ABL fusion and variant Philadelphia chromosome translocations in chronic myelogenous leukemia

Zhang, J; Meltzer, Paul S.; Jenkins, Ronald J.; Guan, Xin Yuan; Trent, Josiah C.

doi:10.1182/blood.v81.12.3365.3365

Cited by 36 publications

(11 citation statements)

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“…One was a commercially obtained whole chromosome 12 painting probe (BRL Life Technologies, Inc.), and the other was a painting probe for 12p generated by us by microdissection of 12p from metaphase cells obtained from phytohemagglutinin-stimulated peripheral blood lymphocytes from a normal individual as described (Meltzer et al, 1992). T h e microdissected DNA was PCR-amplified and biotin-labeled also as described (Zhang et al, 1993).…”

Section: Painting Probes For Detection Of Chromosome Imentioning

confidence: 99%

Molecular cytogenetic analysis of i(12p)‐negative human male germ cell tumors

Rodríguez

Houldsworth

Reuter

et al. 1993

Genes Chromosomes & Cancer

155

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The i( I2p) chromosome has been shown to characterize more than 80% of male germ cell tumors (GCTs) and is an important diagnostic marker. Although recent cytogenetic analyses of GCTs have defined nonrandom chromosome abnormalities in these tumors, no attempt has so far been made to compare i( I2p)-positive and -negative tumors in terms of their cytogenetic, histologic, and clinical features. During a 5-year period, we have ascertained 202 GCTs, of which I17 had clonally abnormal karyotypes. Among the latter, 9 I had one or more copies of i( I2p), whereas 26 lacked an i( I2p). We report here the karyotypic analysis of these 26 i( I2p)-negative GCTs. In this group, nonrandom sites of chromosomal rearrangements included I2p I 3 (9/26) and I p I I -q I I (5126). Comparison of the cytogenetic features of i( I2p)-negative tumors with i( I2p)-positive tumors revealed the only significant difference t o be rearrangements affecting I2pl3 in the former (35%) as compared t o their absence in the latter (3%). Hybridization of metaphase preparations of 9 i( I 2p)-negative tumors with a chromosome I 2 painting probe and with a microdissected I2p painting probe revealed extra copies of chromosome I 2 segments incorporated into marker chromosomes whose composition could not otherwise be resolved by banding analysis; all were shown t o be derived from I2p. These data demonstrate that both i( I 2p)-negative and -positive groups are characterized by an increased copy number of I 2p, which is consistent with a lack of significant clinical or biological difference between them. An increased I2p copy number thus is a specific aberration of significance t o the development of germ cell tumors.

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Section: Painting Probes For Detection Of Chromosome Imentioning

confidence: 99%

Molecular cytogenetic analysis of i(12p)‐negative human male germ cell tumors

Rodríguez

Houldsworth

Reuter

et al. 1993

Genes Chromosomes & Cancer

155

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“…Replacement of the Taq polymerase with a modified bacteriophage T7 DNA polymerase, also known as Sequenase, resulted in larger DNA fragment size than by use of the Taq. This finding is parallel to chromosome microdissection (Bohlander et al, 1992;Guan et al, 1993;Zhang et al, 1993). Since the optimal size of the DNA fragments for the CGH is relatively large (approximately 600-1,200 bp), the use of Sequenase improved clearly both amplification of genomic DNA and quality of CGH hybridization.…”

Section: Discussionmentioning

confidence: 82%

Optimizing DOP-PCR for universal amplification of small DNA samples in comparative genomic hybridization

Kuukasjärvi

Tanner

Pennanen

et al. 1997

Genes Chromosom. Cancer

121

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The standard comparative genomic hybridization (CGH) protocol relies on availability of macroscopic tumor samples, which do not contain too much interfering normal cells. Recently, CGH after universal amplification of genomic DNA with degenerate oligonucleotide primed PCR (DOP‐PCR) has been used to detect genetic aberrations in microdissected tumor specimens. However, owing to the technical difficulties, CGH results of only few microdissected samples have so far been published. We have developed an improved protocol for DOP‐PCR, which includes direct incorporation of fluorochrome‐conjugated nucleotides into the PCR product. Among the four polymerase enzymes tested, ThermoSequenase gave the best yield, with PCR products ranging from 100–4,000 bp. A two‐step PCR‐procedure was used, consisting of a preamplification with low stringency conditions followed by amplification in more stringent conditions. The method was first validated by hybridizing DOP‐PCR‐amplified normal DNA against nick‐translated reference DNA, which showed uniform and even hybridization result for all chromosomes. Comparison of DOP‐PCR CGH to conventional CGH in MCF‐7 breast cancer cell line further indicated that genetic aberrations can be reliably detected after DOP‐PCR amplification. The sensitivity of the DOP‐PCR‐CGH was tested by serial dilution of MCF‐7 DNA. Fifty picograms of sample DNA (corresponding roughly to two MCF‐7 cells) was sufficient for high quality CGH. Experiments with cells microdissected from intraductal breast cancer demonstrated that carcinoma cells from 1 to 2 ducts were sufficient for a successful DOP‐PCR CGH analysis. We conclude that the improved DOP‐PCR‐CGH protocol provides a powerful tool to study genetic aberrations in different histological subpopulations of malignant as well as precancerous lesions. DOP‐PCR also improves the success rate of conventional paraffin‐block CGH, because a poor quality or a too low yield of extracted DNA can be compensated by universal DNA amplification by DOP‐PCR. Genes Chromosom. Cancer 18:94–101, 1997. © 1997 Wiley‐Liss, Inc.

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“…Until recently it was not possible to examine individual interphase cells in the BM or PB for BCR gene rearrangement. However, there are now several approaches for detection of BCR gene rearrangement using FISH (Arnoldus et al, 1990;Tkachuk et al, 1990;Lengauer et al, 1992;Zhang et al, 1993;Zhao et al, 1993;Bentz et al, 1994). We used the sequence-independent amplification method of Bohlander et al (1994) with a different degenerate primer (Telenius et al, 1992) to prepare the BCR-containing YAC (Lengauer et al, 1992) as a probe for detection of BCR gene rearrangement in interphase cells.…”

Section: Discussionmentioning

confidence: 99%

“…T h e BCR YAC contains a small, specific sequence of genomic DNA and therefore provides a more compact, reliable interphase signal (Fig. 1) than probes for the region generated following chromosome microdissection (Zhang et al, 1993) or from somatic cell hybrids (Zhao et al, 1993). Finally, since the YAC probe allows for detection of BCR gene rearrangement using only one color, it can be used simultaneously with other probes detected with a second color.…”

Section: Discussionmentioning

confidence: 99%

Y chromosome loss in chronic myeloid leukemia detected in both normal and malignant cells by interphase fluorescence in situ hybridization

Kirk

VanDevanter

Biberman

et al. 1994

Genes Chromosomes & Cancer

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Loss of the Y chromosome in bone marrow (BM) cells is a normal age-associated event. Y chromosome loss is also observed in the Philadelphia chromosome (Ph) positive BM cells of some patients with chronic myeloid leukemia (CML) in chronic phase, but at a younger age than in normal individuals. While the significance of loss of the sex chromosome in normal males is uncertain, -Y marrow cells are not believed to be of clonal origin. However, because CML is a clonal disease, CML sub-populations with Y loss may constitute a disease-related sub-clone. We used a PCR-amplified yeast artificial chromosome containing the BCR gene region for single color interphase analysis of BCR rearrangement by fluorescence in situ hybridization (FISH). Then, using two color FISH, with one fluorochrome detecting the BCR gene region and the other detecting Y chromosome repeat sequences, we surveyed peripheral and BM Y loss in both normal Ph- (BCR not disrupted) and CML Ph+ (BCR rearranged) interphase nuclei of two patients with Y loss in Ph positive cells observed by metaphase analysis. -Y was seen in a proportion of Ph+ cells in both cases, and the proportion matched that seen in Ph- cells, indicating that Y loss is probably sporadic in both normal and CML populations, and that the propensity for Y loss in normal BM cells may be a phenotype that can be retained by malignant cells in CML.

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Application of chromosome microdissection probes for elucidation of BCR- ABL fusion and variant Philadelphia chromosome translocations in chronic myelogenous leukemia

Cited by 36 publications

References 0 publications

Molecular cytogenetic analysis of i(12p)‐negative human male germ cell tumors

Molecular cytogenetic analysis of i(12p)‐negative human male germ cell tumors

Optimizing DOP-PCR for universal amplification of small DNA samples in comparative genomic hybridization

Y chromosome loss in chronic myeloid leukemia detected in both normal and malignant cells by interphase fluorescence in situ hybridization

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