Analysis of genes involved in response to doxorubicin and a GD2 ganglioside-specific 14G2a monoclonal antibody in IMR-32 human neuroblastoma cells.
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.
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Irena
I
Horwacik
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Małgorzata
M
Durbas
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Elżbieta
E
Boratyn
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Anna
A
Sawicka
Department of Cellular Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Paulina
P
Węgrzyn
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Sylwia
S
Krzanik
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Anna
A
Górka
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Joanna
J
Drożniak
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Ewa
E
Augustyniak
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Aleksandra
A
Kowalczyk
Laboratory of Molecular Genetics and Virology, Faculty of Biochemistry, Biophysics and Biotechnology, The Jagiellonian University, Kraków, Poland.
Hanna
H
Rokita
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D045744:Cell Line, Tumor / Q000187:drug effects; D016000:Cluster Analysis; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000737:chemistry*; D005732:Gangliosides / Q000737:chemistry*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D051379:Mice; D024101:Mitochondrial Proteins / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000473:pathology*; D009687:Nuclear Proteins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D015513:Oncogene Proteins / Q000378:metabolism; D012984:Software; D019772:Topotecan / Q000737:chemistry; D059467:Transcriptome
D000903:Antibiotics, Antineoplastic; D000911:Antibodies, Monoclonal; D005732:Gangliosides; C459801:MYCN protein, human; D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; D000071447:N-Myc Proto-Oncogene Protein; C501824:NDUFAF2 protein, human; D009687:Nuclear Proteins; D015513:Oncogene Proteins; C019403:ganglioside, GD2; D019772:Topotecan; D004317:Doxorubicin