Notch1 siRNA影响Notch-Wnt信号通路对人成骨肉瘤细胞增殖的抑制作用  

RIPK4 participates in osteosarcoma cell proliferation by regulating the Notch-Wnt signaling pathway

作　　者：郝俊龙[1] 杨凯[1] 王亚鹏 王旭[1] 齐进[1] 王拴科[1] 汪静[1] HAO Junlong;YANG Kai;WANG Yapeng;WANG Xu;QI Jin;WANG Shuanke;WANG Jing(Department of Orthopaedics,Second Hospital of Lanzhou University,Lanzhou 730030,Gansu Province,China)

机构地区：[1]兰州大学第二医院骨肿瘤科,甘肃兰州730030

出　　处：《中国癌症杂志》2018年第8期567-576,共10页China Oncology

基　　金：甘肃省科技厅创新团队项目(1308RJIA004)

摘　　要：背景与目的:骨肉瘤是骨肿瘤中最常见的原发性恶性肿瘤。迄今为止,骨肉瘤发生、发展的相关分子机制尚不明确,临床上也没有针对性的有效治疗药物。该研究探索Notch1蛋白在骨肉瘤中的作用及与影响骨肉瘤发生、发展的Notch-Wnt信号通路之间的关系。方法:采用免疫组织化学染色法检测骨肉瘤组织中Notch1的表达;采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法与流式细胞术检测靶向Notch1si RNA转染的人骨肉瘤细胞MG-63与U-2 OS凋亡的影响,实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)、蛋白质印迹法(Western bolt)检测转染后骨肉瘤细胞内Notch与Wnt信号通路中相关蛋白及RIPK4蛋白的表达变化。结果:Notch1在人骨肉瘤活检组织中的表达呈阳性,同时Notch1 si RNA能够显著抑制人骨肉瘤MG-63和U-2 OS细胞的增殖,并促进骨肉瘤细胞发生凋亡。此外诱导降低或缺失Notch1后,Notch与Wnt信号通路中相关蛋白及RIPK4蛋白表达明显减少。结论:si RNA-Notch1脂质体转染骨肉瘤细胞后,可影响Notch1-Wnt信号通路并降低促增殖分子RIPK4的表达,发挥抑制肿瘤细胞生长的作用。Background and purpose:Osteosarcoma is the most common primary malignancy in bone tumors.So far,the molecular mechanism of the development of osteosarcoma is not clear,and there is no effective therapeutic drug in clinic.This study explored the relationship between Notch1 protein and osteosarcoma and whether their influence involved in the occurrence and development of Notch-Wnt signal pathways.Methods:The expression levels of Notch1 mRNA and protein in tissue specimens were detected by immunohistochemistry,real-time fluorescent quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Transfection of siRNA interference Notch1 gene was carried out in MG-63 and U-2 OS osteosarcoma cell lines respectively to achieve reduction of Notch1 osteosarcoma cells.Osteosarcoma cell proliferation and apoptosis were determined by MTT test and flow cytometry.RTFQ-PCR and Western blot were used to detect the protein content changes of Notch and Wnt signaling pathway.Results:Notch1 in the tissue of bone sarcoma obtained by biopsy was positive.Notch1 siRNA significantly inhibited MG-63 and U-2 OS cells proliferation,and promoted apoptosis.It showed that the protein expression decreased significantly in Notch and Wnt signaling pathway after transfection of osteosarcoma cells with Notch1-siRNA.Conclusion:Notch1 plays a role in promoting proliferation by influencing the Notch-Wnt signaling pathway in the occurrence and development of osteosarcoma.

关 键 词：NOTCH1 MG-63细胞 U-2 OS细胞 Notch-Wnt信号通路 

分 类 号：R738.1[医药卫生—肿瘤]