CAJ | 학술논문

目的以自主建立的有骨髓转移的神经母细胞瘤细胞系(QDDQNM1)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(QDDQ-NM1)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P65 mRNA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48 h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.
목적 이자주건립적유골수전이적신경모세포류세포계(QDDQNM1)위대상,초보탐토NF-κB통로재신경모세포류(NB)전이과정중적작용급기궤제.방법 본실험장세포계(QDDQ-NM1)분위3조,즉:종류배사인자간예조(Ⅰ조)、정상대조조(Ⅱ조)화PDTC간예조(Ⅲ조).이용RT PCR검측NF-κB통로주요단백P65 mRNA급기하유단백추화인자수체4(CXCR4) mRNA적표체；Western-blot법검측P65단백급CXCR4단백적표체；Transwell소실검측각조간류세포적전이침습능력.결과 Ⅰ、Ⅱ、Ⅲ조P65 mRNA상대함량분별위0.7211±0.2233、0.6002±0.1822화0.5553±0.0904,CXCR4 mRNA상대함량분별위0.4128±0.0176、0.3045±0.1910화0.1829±0.0091,각조간차이유통계학의의(P＜0.05).Ⅰ、Ⅱ、Ⅲ조P65단백상대함량분별위0.9026±0.0375、0.8340±0.0379화0.8007±0.0793,CXCR4단백상대함량분별위0.9472±0.0193、0.9091±0.0216화0.6025±0.0384,각조간차이유통계학의의(P＜0.05).류세포배양48 h후,Ⅰ、Ⅱ、Ⅲ조중천과ECM효적세포수분별위55.8000±2.7749、46.6000±5.9414화30.2000±7.4967,각조간차이유통계학의의(P＜0.05).결론 NF-κB통로통과조절기하유단백CXCR4적표체래증강NB세포적침습능력.통과억제NF-κB통로래강저NB세포적침습전이능력,유망성위치료NB적신도경.
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-α treatment group; group Ⅱ,the control group; and group Ⅲ,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 QDDQ-NM1cells.Results The mRNA expression of P65 of the QDDQ-NM1 cells was significantly different between the 3 groups (0.721 1 ± 0.2233,0.6002 ± 0.1822 and 0.5553 ± 0.0904,respectively,P＜0.05).The mR NA expression of CXCR4 of the QDDQ-NM1cells of group Ⅰ,group Ⅱ and group Ⅲ was also significantly different (0.4128 ± 0.0176,0.3045 ± 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).Conclusions NF-kappa B pathway is involved in neuroblastoma invasion and metastasis