摘要
目的研究Dickkopf-1（DKK-1）在儿童神经母细胞瘤（NB）中的表达及其意义。方法运用免疫组织化学方法检测DKK-1在42例NB，19例神经节细胞瘤及15例正常肾上腺组织中的表达水平。运用实时荧光定量PCR（qRT-PCR）、Western blotting检测DKK-1在分化程度依次增高的3株NB细胞株IMR-32、SK-N-SH和SH-SY5Y中的mRNA及蛋白表达水平。结果免疫组织化学研究发现DKK-1在正常肾上腺组织中呈强阳性表达（免疫组织化学评分为1．912±0．047），在神经节细胞瘤组织中呈弱阳性表达（免疫组织化学评分为1．214±0．095），在NB组织中呈阴性表达（免疫组织化学评分为0．371±0．293）。神经节细胞瘤和NB中DKK-1的表达量分别与正常对照组相比较，差异均有统计学意义（all P〈0．001）。在3种组织中，不同组间DKK-1表达量的差异有统计学意义（F=211．375，P〈0．001）。qRT-PCR研究发现DKK-1在IMR-3、SK-N-SH和SH-SY5Y中平均相对表达量分别为（25．323±8．651）％、（58．852±10．223）％和（75．385±9．381）％，均呈低表达水平，不同组间DKK-1表达差异有统计学意义（F=57．628，P〈0．001）。Western blotting研究发现在IMR-32、SK-N-SH和SH-SY5Y中DKK-1蛋白表达相对灰度值依次增高，分别为0．093±0．016、0．397±0．051和0．411±0．062，不同组间蛋白表达差异有统计学意义（F=156．783，P〈0．001）。结论DKK-1在NB组织及细胞株中均呈低表达水平，且DKK-1在NB中的表达可能与该肿瘤的分化密切相关，提示其或许可以作为一个特殊的分子标记物用于NB的早期诊断及预后监测。
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-SYSY 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.
出处
《中华小儿外科杂志》
CSCD
2016年第9期643-647,共5页
Chinese Journal of Pediatric Surgery
基金
国家自然科学基金（81502187）
