Abstract:

Objective To elucidate the inhibitory effect of interferon-γ（IFN-γ） on the proliferation of neuroblastoma cells and the protentral gene signature of IFN-γ and the relationship between the expression of gene signature of IFN-γ in the neuroblastoma cells and its adverse prognosis， and to clarify the effect of IFN-γ and its gene signture in the neuroblastoma. Methods The SK-N-BE（2）（proto-oncogene N-MYC amplification type） and SH-SY5Y （proto-oncogene N-MYC non-amplification type） neuroblastoma cells were selected and treated with different concentrations （0， 500， 750， 1 000 and 1 500 μg·L^-1） of IFN-γ for 24 h， followed by cell proliferation assays using cell counting kit-8 （CCK-8）. Transcriptome sequencing was then performed to identify the gene signature of IFN-γ. Additionally， the tissue microarrays from 23 cases of neuroblastoma and 6 cases of normal adrenal gland samples were collected， immunohistochemistry （IHC） analysis was used to to detect the expression of gene signature of IFN-γ. Based on the expression levels of gene signature of IFN-γ， the samples were divided into SULT2B1 low and high expression groups. The correlation between the expression of gene signature of IFN-γ and poor prognosis of the patients was analyzed. Results The CCK-8 assay results showed that as IFN-γ concentration increased，the proliferation of SK-N-BE（2） cells was significantly inhibited （P<0.01）， with inhibitory rates of SK-N-BE（2） cells in four groups were 6.73%， 6.77%，7.67%， and 9.19%， respectively. In contrast， the proliferation rate of SH-SY5Y cells were significantly increased with the increase of IFN-γ concentrations （P<0.01）， and the proliferation rates of SH-SY5Y cells in four groups were 46.80%， 79.19%， 70.30%， and 72.33%， respectively. Transcriptome sequencing identified hydroxysteroid sulfotransferase 2B1 （SULT2B1） as a potential gene signature of IFN-γ. The IHC analysis results showed the expression amount of SULT2B1 protein in neuroblastoma tissues was increased. The clinical data analysis results revealed significant differences in age（Z=-2.618， P=0.018）， lymphnode metastasis （χ²=4.439， P=0.035）， and distant metastasis （χ²=5.856， P=0.016） between low and high SULT2B1 expression groups. Conclusion IFN-γ can inhibit the proliferation of SK-N-BE（2） cells while promoting the proliferation of SH-SY5Y cells. SULT2B1 is a potential gene signature of IFN-γ， and its expression is upregulated in neuroblastoma tissue. SULT2B1 high expression is significantly associated with poor prognosis in the neuroblastoma patients.

Key words: Neuroblastoma, Interferon-γ, Hydroxysteroid sulfotransferase 2B1, Oncogene, Tumor metastasis