摘要
目的以自主建立的有骨髓转移的神经母细胞瘤细胞系（QDDQ-NMI）为对象，初步探讨NF-κB通路在神经母细胞瘤（NB）转移过程中的作用及其机制。方法本实验将细胞系（QDDQNM1）分为3组，即：肿瘤坏死因子干预组（Ⅰ组）、正常对照组（Ⅱ组）和PDTC干预组（Ⅲ组）。利用RT-PCR检测NF-κB通路主要蛋白P65 mRNA及其下游蛋白趋化因子受体4（CXCR4）mRNA的表达；Western-blot法检测P65蛋白及CXCR4蛋白的表达；Transwell小室检测各组间瘤细胞的转移侵袭能力。结果Ⅰ、Ⅱ、Ⅲ组P65mRNA相对含量分别为0．7211±0．2233、0．6002±0．1822和0．5553±0．0904，CXCR4 mRNA相对含量分别为0．4128±0．0176、0．3045±0．1910和0．1829±0．0091，各组间差异有统计学意义（P〈0．05）。Ⅰ、Ⅱ、Ⅲ组P65蛋白相对含量分别为0．9026±0．0375、0．8340±0．0379和0．8007±0．0793，CXCR4蛋白相对含量分别为0．9472±0．0193、0．9091±0．0216和0．6025±0．0384，各组间差异有统计学意义（P〈0．05）。瘤细胞培养48h后，Ⅰ、Ⅱ、Ⅲ组中穿过ECM胶的细胞数分别为55．8000±2．7749、46．6000±5．9414和30．2000±7．4967，各组间差异有统计学意义（P〈0．05）。结论NF-κB通路通过调节其下游蛋白CXCR4的表达来增强NB细胞的侵袭能力。通过抑制NF-κB通路来降低NB细胞的侵袭转移能力，有望成为治疗NB的新途径。
Objective To investigate the role of NF-kappa B pathway in neuroblastoma invasion and metastasis. Methods QDDQ-NM1 cells, an independent neuroblastoma cell line, were used in this study. The cells were divided into 3 groups according to the treatment to the cells： group I , TNF-Ⅰ treatment group; group Ⅱ, the control group; and groupⅢ, the PDTC treatment group. The mRNA expressions of P65 and the NF-KB pathway downstream protein CXCR4 were investigated by RT-PCR. The protein expressions of P65 and CXCR4 were studied by western blotting. Furthermore,a Transwell invasion chamber was used to study the invasion and metastasis of QDDQI-NMlcells. Results The mRNA expression of P65 of the QDDQ-NM1 cells was significantly different between the 3 groups （0. 7211 ± 0. 2233,0. 6002 ± 0. 1822 and 0. 5553 ± 0. 0904, respectively, P〈0. 05）. The mRNA expression of CXCR4 of the QDDQ-NM1cells of group Ⅰ, group Ⅱ and group Ⅲ was also significantly different （0. 4128 ± 0. 1）176,0. 3045 ± 0. 1910 and 0. 1829± 0. 0091, respectively, P〈0.05）. The protein level of P65 of the 3 groups was 0. 9026 ± 0. 0375,0. 8340 ± 0. 0379 and 0. 8007 ± 0. 0793,respectively. The protein level of CXCR4 of the 3 groups was 0. 9472 ± 0. 0193,0. 9091 ± 0. 0216 and 0. 6025 ±0. 0384, respectively. The protein levels of P65 and CXCR4 were significantly different between the 3 groups （P〈0. 05）. The Transwell assay revealed that, after 48 hours of culture, the number of QDDQ-NM1 cells passing through ECM membrane between the 3 groups was significantly different （55. 8000 ± 2. 7749, 46. 6000 ± 5. 9414 and 30. 2000 ± 7. 4967 respectively, P〈0. 05）. Conelusions NF-kappa B pathway is involved in neuroblastoma invasion and metastasis
出处
《中华小儿外科杂志》
CSCD
北大核心
2013年第3期195-198,共4页
Chinese Journal of Pediatric Surgery
