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Regulation of acute phase reaction by transforming growth factor β in cultured murine hepatocytes

100%

Rokita H., Szuba K.

Acta Biochimica Polonica

1991

vol. 38

issue 2

241-249

New method for quantitative analysis of GD2 ganglioside in plasma of neuroblastoma patients

73%

Czaplicki D., Horwacik I., Kowalczyk A., Wieczorek A., Bolek-Marzec K., Balwierz W., Kozik A., Rokita H.

issue 3

423-431

Neuroblastoma, the most common extracranial solid tumour of childhood, is a malignancy of unknown origin and non-specific symptoms. One of the markers of the disease is GD2 ganglioside (disialoganglioside), which is abundantly expressed on the surface of neuroblastoma cells. Gangliosides are known to be shed by tumour cells and this phenomenon can be significant in cancer progression as they inhibit a number of immune responses both in vitro and in vivo. In search for novel markers useful in monitoring and prognosis of neuroblastoma, we developed and validated a new quantitative method of GD2 ganglioside analysis in human blood plasma. We evaluated the level of gangliosides in blood serum of 34 neuroblastoma patients using high-performance liquid chromatography. The technique was used to detect fluorescently labelled oligosaccharides derived from serum glycosphingolipids by enzymatic digestion with ceramide glycanase. The developed method allowed determination of GD2 concentrations at the picomole level and required only 40 µl of plasma, which should be particularly useful when the quantity of clinical material is limiting. Moreover, this method can be applied to study concentration of other gangliosides, as shown for GD3 ganglioside. Analysis of plasma samples from the 34 neuroblastoma patients did not reveal any correlations between the concentration of GD2 ganglioside and clinical parameters, including the results of therapy; it showed, however, that the concentration of GD2 ganglioside in the plasma of neuroblastoma patients decreased substantially in the course of treatment.

Analysis of genes involved in response to doxorubicin and a GD2 ganglioside-specific 14G2a monoclonal antibody in IMR-32 human neuroblastoma cells

60%

Horwacik I., Durbas M., Boratyn E., Sawicka A., Węgrzyn P., Krzanik S., Górka A., Drożniak J., Augustyniak E., Kowalczyk A., Rokita H.

2015

vol. 62

issue 3

423-433

Neuroblastoma is the most common extra-cranial solid tumor of childhood and it is characterized by the presence of a glycosphingolipid, GD2 ganglioside. Monoclonal antibodies targeting the antigen are currently tested in clinical trials. Additionally, several research groups reported results revealing that ganglioside-specific antibodies can affect cellular signaling and cause direct cytotoxicity against tumor cells. To shed more light on gene expression signatures of tumor cells, we used microarrays to analyze changes of transcriptome in IMR-32 human neuroblastoma cell cultures treated with doxorubicin (DOX) or a mouse monoclonal antibody binding to GD2 ganglioside 14G2a (mAb) for 24 h. The obtained results highlight that disparate cellular pathways are regulated by doxorubicin and 14G2a. Next, we used RT-PCR to verify mRNA levels of selected DOX-responsive genes such as RPS27L, PPM1D, SESN1, CDKN1A, TNFSF10B, and 14G2a-responsive genes such as SVIL, JUN, RASSF6, TLX2, ID1. Then, we applied western blot and analyzed levels of RPS27L, PPM1D, sestrin 1 proteins after DOX-treatment. Additionally, we aimed to measure effects of doxorubicin and topotecan (TPT) and 14G2a on expression of a novel human NDUFAF2 gene encoding for mimitin protein (MYC-induced mitochondrial protein) and correlate it with expression of the MYCN gene. We showed that expression of both genes was concomitantly decreased in the 14G2a-treated IMR-32 cells after 24 h and 48 h. Our results extend knowledge on gene expression profiles after application of DOX and 14G2a in our model and reveal promising candidates for further research aimed at finding novel anti-neuroblastoma targets.

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3 Acta Biochimica Polonica

1 2015

1 2009

1 1991

3 Rokita H.

2 Horwacik I.

2 Kowalczyk A.

1 Augustyniak E.

1 Balwierz W.

1 Bolek-Marzec K.

1 Boratyn E.

1 Czaplicki D.

1 Drożniak J.

1 Durbas M.

1 Górka A.

1 Kozik A.

1 Krzanik S.

1 Sawicka A.

1 Szuba K.

1 Wieczorek A.

1 Węgrzyn P.

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Regulation of acute phase reaction by transforming growth factor β in cultured murine hepatocytes

New method for quantitative analysis of GD2 ganglioside in plasma of neuroblastoma patients

Analysis of genes involved in response to doxorubicin and a GD2 ganglioside-specific 14G2a monoclonal antibody in IMR-32 human neuroblastoma cells