Abstract
Objective To explore the expression and significance of DKK-1 in neuroblastoma (NB). Methods Immunohistochemical staining (IHC) was employed for detecting the expression of DKK-1 in 42 NB tissues and 19 gangliocytoma tissues. And 15 normal adrenal tissues were used as a positive control. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot were used for detecting mRNA and protein expressions of DKK-1 in 3 NB cell lines of IMR-32, SK-N-SH and SH-SY5Y. And HEK293 was used as a positive control. Results The results of IHC showed that mean expression scores for DKK-1 were 1.912±0.047, 1.214±0.095 and 0.371±0.293 in normal adrenal gland tissues, ganglioneuroma and NB cells respectively. DKK-1 protein was down-regulated in ganglioneuroma and NB cells as compared to normal adrenal gland tissues (all P<0.001). Multiple comparison indicated that the differences of protein expression for DKK-1 were statistically significant within 3 tissues (F=211.375, P<0.001). The results of qRT-PCR showed that the relative mean mRNA expression of DKK-1 was (25.323±8.651)%, (58.852±10.223)% and (75.385±9.381)% in IMR-32, SK-N-SH and SH-SY5Y cells respectively. DKK-1 mRNA level became down-regulated in all 3 NB cell lines as compared to HEK293 (all P<0.001). Multiple comparison indicated that the differences of DKK-1 expression were statistically significant among 3 NB cell lines (F=57.628, P<0.001). Western blot showed that the relative mean protein expression of DKK-1 was 0.093±0.016, 0.397±0.051 and 0.411±0.062 in IMR-32, SK-N-SH and SH-SY5Y cells respectively. DKK-1 protein became down-regulated in 3 NB cell lines as compared to HEK293 (all P<0.001). Multiple comparison showed statistically significant differences of DKK-1 expression among 3 NB cell lines (F=156.783, P<0.001). Conclusions DKK-1 becomes down-regulated in NB tissues and cells and it has close relationship with differentiation. Thus DKK-1 may serve as a potential molecular marker for early diagnosis and prognostic monitoring of NB. Key words: Neuroblastoma; Gene expression; Comparative study
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