神经母细胞瘤不同侵袭能力细胞株的建立及研究

Exploring the correlation between expression of CXCR4 and in-vitro metastatic capacity in neuroblastoma cell sublines taken from the same patient

目的建立同一患儿神经母细胞瘤（NB，neuroblastoma）细胞系来源的不同侵袭能力的细胞株，探讨CXCR4（趋化因子受体4）的表达水平对神经母细胞瘤细胞株体外侵袭能力的影响。方法采用有限稀释法，从人神经母细胞瘤细胞系（QDDQ-NM）中获得5株CXCR4表达水平不同的单克隆细胞株，应用免疫组化和Rrr_PCR检测细胞株CXCR4蛋白和CXCR4mRNA的表达水平，Tran—swell体外趋化侵袭实验检测5株细胞株趋化侵袭能力的差异。结果CXCR4在细胞株QDDQ-NM1中的表达（94．20±4．19）明显高于细胞株QDDQ-NM5（79．58±5．10），P〈0．05；CXCIⅥmRNA在QDDNM1中的表达（2．26±0．23）明显高于细胞株QDDKvNM5（1．83±0．08），Pd0．05。QD—NM1的体外趋化侵袭能力（54±4）明显强于QDDQ-NM5（45±1．58），P〈0．05，其与CXCR4蛋白和CXCR4mRNA表达水平呈正相关。结论同一患儿神经母细胞瘤细胞系中的细胞具有异质性，不同细胞株的侵袭能力不同，神经母细胞瘤细胞株高度的体外侵袭能力与CXCR4的高表达呈高度相关性。

Objective To explore the correlation between expression of CXCR4 and in-vitro me- tastatic capacity in neuroblastoma cell sublines taken from the same patient. Methods Neuroblastoma cell sublines were selected from human neuroblastoma cell line （QDDQ-NM） cultured by ourselves using unicell cloning technique,the expressions of protein and mRNA of CXCR4 were detected by immunohistochemistry and RT-PCR, the pulmonary metastatic potential was detected by the Transwell in-vitro invasion assay. Results The two neuroblastoma cell sublines （QDDQ-NM 1, QDDQ-NM 5） were ,obtained, The expression of protein CXCR4 of sublines QDDQ-NM 1 was 94. 20 ± 4. 19 and was significantly higher than in QDDQ-NM 5 （79. 58 ±5. 10） ,P〈0. 01. The expression of CXCR4 mRNA in QDDNM 1 （2. 26 ± 0. 23） was also significantly higher than QDDQ-NM 5 （1.83±0.08）, P〈 0. 01. The in-vitro metastatic potentials of QDDO-NM 1 （54± 4）was significantly higher than QDDQ NM 5 （45 ±1.58）, P〈0. 01, which showed high correlation with the expression of CXCR4 and CXCR4 mRNA. Conclusions High expression of CXCR4 is assiociated with high in-vitro metastatic potential of human neuroblastoma cell sublines.