HA117 inhibits the differentiation via regulation of DPF3 in neuroblastoma

Abstract

Objective To investigate the expression and signification of HA117 and DPF3, as well as the effects of them on the differentiation in neuroblastoma. Methods Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry were employed to detect the expression of HA117 and DPF3 mRNA and protein expression in 35 neuroblastomas (NB), 8 ganglioneuroblastomas (GNB) and 10 ganglioneuromas(GN). HA117 overexpression and control groups were established by lentivirus transduction in SH-SY5Y. qRT-PCR and Western blot were used to detect HA117 and DPF3 mRNA and protein expression level in cells. After SH-SY5Y cells of HA117 overexpression group and control group were induced with 10μmol/L retinoic acid(RA) for 7 days, cell morphology was observed by phase contrast microscop, immunofluorescence was performed to examine MAP2 expression, Western blot was employed for detecting the expression of MAP2 and NSE. Results The RNA levels of HA117 in NB were significantly higher than those in GNB/GN, conversely(30.14±15.96 VS 1.03±0.37, P<0.05), the mRNA levels of DPF3 in NB were significantly lower than those in GNB/GN(0.81±0.17 VS 3.14±1.09, P<0.05). The immunohistochemical staining of DPF3 was strongly positive in GNB/GN, weakly positive in differentiating NB and negative in undifferentiated or poorly differentiated NB. The mRNA and protein expression levels were significantly negatively regulated by HA117 overexpression in SH-SY5Y cells. After induced with RA, cells in control group growed slowly and differentiated with synapses. The protein levels of MAP2 and NSE in HA117 overexpression group were significantly lower than those in control group(P<0.05). Conclusions The expression of HA117 and DPF3 are related with the differentiation of neuroblastoma, HA117 may inhibits the differentiation of neuroblastoma through regulation of DPF3. Key words: Neuroblastoma; Gene expression; Differentiation