Inositide-specific Phospholipase C beta1 gene deletion in the progression of Myelodisplastic Syndrome to Acute Myeloid Leukemia

Myelodysplastic syndrome (MDS) is an adult hematological disease that evolves into acute myeloid leukemia (AML) in about 30% of the cases. The availability of a highly specific probe moved us to perform in patients affected with MDS/AML, associated with normal karyotype, painting and fluorescence in situ hybridization (FISH) analysis aimed to check the inositide-specific phospholipase C (PI-PLC) beta1 gene, a player in the control of some checkpoints of the cell cycle. Here we present a preliminary observation in which FISH analysis disclosed in a small group of MDS/AML patients with normal karyotype the monoallelic deletion of the PI-PLCbeta1 gene. On the contrary, PI-PLC beta4, another gene coding for a signaling molecule, located on 20p12.3 at a distance as far as less than 1Mb from PI-PLCbeta1, is unaffected in MDS patients with the deletion of PI-PLC beta1 gene, hinting at an interstitial deletion. The MDS patients, bearing the deletion, rapidly evolved to AML. The data suggest the possible involvement of PI-PLCbeta1 in the progression of the disease and pave the way for a larger investigation aimed at identifying a possible high-risk group among MDS patients with a normal karyotyp

Inositide-specific Phospholipase C beta1 gene deletion in the progression of Myelodisplastic Syndrome to Acute Myeloid Leukemia

Myelodysplastic syndrome (MDS) is an adult hematological disease that evolves into acute myeloid leukemia (AML) in about 30% of the cases. The availability of a highly specific probe moved us to perform in patients affected with MDS/AML, associated with normal karyotype, painting and fluorescence in situ hybridization (FISH) analysis aimed to check the inositide-specific phospholipase C (PI-PLC) beta1 gene, a player in the control of some checkpoints of the cell cycle. Here we present a preliminary observation in which FISH analysis disclosed in a small group of MDS/AML patients with normal karyotype the monoallelic deletion of the PI-PLCbeta1 gene. On the contrary, PI-PLC beta4, another gene coding for a signaling molecule, located on 20p12.3 at a distance as far as less than 1Mb from PI-PLCbeta1, is unaffected in MDS patients with the deletion of PI-PLC beta1 gene, hinting at an interstitial deletion. The MDS patients, bearing the deletion, rapidly evolved to AML. The data suggest the possible involvement of PI-PLCbeta1 in the progression of the disease and pave the way for a larger investigation aimed at identifying a possible high-risk group among MDS patients with a normal karyotyp

Molecular and cytogenetic characterization of stem-like cells in human papillary thyroid carcinoma B-CPAP cell line

Several kinds of tumors and cancer cell lines may contain a subpopulation of cancer stem cells (CSCs) (Setoguchi et al., 2004). In this study, we attempted to enrich CSCs from the PTC-derived B-CPAP cell line using the sphere culture system, in order to molecularly and cytogenetically characterize them. B-CPAP cell lines were able to grow as floating spheres (thyrospheres) in serum-free medium supplemented with EGF and bFGF. Thyrospheres were able to expand for more than 20 passages, with exponentiallyincreasing sphere-forming efficiency and self-renewal, up to passage 8. RT-PCR analysis showed that stem cell markers Oct 4 and Nanog were expressed at the same level in both B-CPAP adherent cells and thyrospheres, whereas ABCG2 and ALDH1A1 were expressed more in thyrospheres. Moreover, thyroid differentiation markers TTF1 and PAX8 were both expressed in B-CPAP thyrospheres and adherent cells, although TTF1 expression was reduced in thyrospheres. Expression of ALDHA1 protein was present only in thyropspheres, while CK19 protein, a differentiation epithelial marker, was expressed only in adherent cells. Spectral karyotyping imaging investigation showed a more complex karyotype, with additional chromosomal rearrangements in thyrospheres as compared to adherent cells, in keeping with possible higher genomic instability of CSCs. Stem-like cells from thyrospheres were isolated by flow cytometry cell sorting assay, using the lipophylic fluorescent dye PKH26. Two cell populations were isolated: the brightest (PKH26 High) making up putative stem cells and the dimmest (PKH26 Low) representing proliferating cells. PKH26 High retained the ability to form secondary spheres and showed increased expression of stem cell markers as compared to PKH26 Low. As expected, differentiated markers were expressed more in PKH26 Low. As a whole, our data suggest that the B-CPAP cell line contains cells with stem-like features, including genomic instability. Supported by Regione Autonoma Sardegna, POR -FSE 2007–2013

A quarter of men with idiopathic oligo-azoospermia display chromosomal abnormalities and microdeletions of different types in interval 6 of Yq11.

Cytogenetic investigations and molecular analysis of the Y chromosome by the polymerase chain reaction amplification of sequence-tagged sites (STS-PCR) technique were performed in 126 patients affected by idiopathic oligo-azoospermia following accurate selection of cases. Seventeen patients evidenced an abnormal karyotype. Fourteen patients with a normal karyotype had microdeletions of the Y chromosome within interval 6. In azoospermic patients microdeletions were scattered along different subintervals, while in oligozoospermic patients they were clustered in subinterval 6E. The size of the deletion was not apparently related to the severity of the disease. These results suggest that cytogenetic analysis and the STS-PCR technique can detect a genetic cause of infertility in about one-quarter of patients with idiopathic oligo-azoospermia