RNAi沉默Notch1基因对胃癌BGC-823细胞增殖和迁移及侵袭能力的影响  

Influence of silencing Notch1 on proliferation, migration and invasion of BGC-823 gastric cancer cells

作　　者：赵恩昊[1] 金鑫[1] 沈志勇[1] 刘骅[1] 曹晖[1] 

机构地区：[1]上海交通大学附属仁济医院普通外科,200127

出　　处：《中华胃肠外科杂志》2012年第12期1296-1300,共5页Chinese Journal of Gastrointestinal Surgery

摘　　要：目的研究Notch1基因在胃癌细胞增殖、迁移和侵袭过程中的作用。方法采用RT-PCR和Western blot分别检测不同的胃癌细胞（SGC-7901、BGC．823、MKN-45和MKN-28）和正常胃黏膜上皮细胞（GES-1）Notch1的表达水平。构建RNAi片段靶向抑制Notch1高表达的胃癌细胞株，采用CCK8实验和迁移侵袭实验检测RNAi对胃癌细胞体外的增殖、迁移和侵袭能力的影响。结果胃癌BGC-823细胞中Notch1mRNA和蛋白均呈现高表达，RNAi片段转染后BGC-823细胞的Notch1 mRNA和蛋白表达水平明显下降。CCK8实验显示，从转染第3天起，RNAi组细胞生长速度明显减缓（F=4．644，P=0．046）；迁移实验结果显示，RNAi组的穿膜细胞[（30．1＋3．1）个]显著低于阴性对照组[（44．5±3．2）个]和空白对照组[（46．1±2．6）个]；侵袭实验的结果也显示，RNAi组的穿膜细胞数[（22．9±4．5）个]显著低于阴性对照组[（37．3±4．2）个]和空白对照组[（37．8±3．7）个]，各组间的差异有统计学意义（P〈0．05）。结论Notch1基因表达的降低可以抑制胃癌BGC-823细胞的增殖，降低细胞的迁移和侵袭能力，Notch1可能与胃癌的发生、发展和转移有着密切的关系。Objective To investigate the role of Notchl in the proliferation, migration and invasion of gastric cancer cells. Methods Notch1 expression was detected by RT-PCR and Western blot in four gastric cancer cell lines （SGC-7901, BGC-823, MKN-45 and MKN-28）. The RNAi segments targeting Notch1 were designed and transfected into cell lines with the highest level of Notch1. The transcription and translation level of Notch1 expression was detected by real-time PCR and Western blot. The proliferation of transfected cells were determined by CCK8 test. Meanwhile, the migration and invasive ability of transfected tumor cells were detected by Transwell test. Results Gastric cell line BGC-823 was selected with the highest expression of Notchl at both mRNA level and protein level. The expression of Notchl was significantly reduced after the transfection. The CCK8 test showed the growth rate was significantly reduced in RNAi group after two days （F=4.644, P=0.046）. In migration test, the number of cells crossing through chambers in RNAi group （30.1±3.1） was significantly lower than that in the negative control group（44.5±3.2） and in the mock group（46.1±2.6）（P〈0.05）. In invasion test, the number of cells crossing through chambers in RNAi group （22.9±4.5） was also significantly lower than that in the negative control group （37.3±4.2） and in the mock group （37.8±3.7） （P〈0.05）. Conclusion Down-regulation of Notchl inhibits the proliferation and reduces the ability of migration and invasion in gastric cell line BGC-823, suggesting that Notchl may play an important role in the gastric carcinogenesis and metastasis.

关 键 词：胃肿瘤细胞 NOTCH1基因 RNA干扰 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]