沉默乙酰肝素酶基因对腺样囊性癌细胞侵袭和迁移能力的影响  

Role of heparanase gene silence on the invasion and migration of human salivary adenoid cystic carcinoma cells

作　　者：王新[1] 龚中坚[1] 於俊[1] 

机构地区：[1]南京医科大学附属无锡市人民医院口腔科,江苏无锡214023

出　　处：《南京医科大学学报（自然科学版）》2013年第10期1383-1387,共5页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的:探讨siRNA干扰沉默乙酰肝素酶(heparanase,HPA)基因对人腺样囊性癌SACC-M细胞侵袭和迁移能力的影响。方法:根据人HPA基因序列设计并合成质粒表达载体转染至人涎腺腺样囊性癌(salivary adenoid cystic carcinoma,SACC)细胞,采用逆转录-聚合酶链反应(RT-PCR)和Western blot检测转染前后HPA mRNA和蛋白的表达变化,选出对HPA沉默效果最佳的质粒,获得稳定表达细胞株。通过划痕损伤实验和Transwell小室细胞侵袭实验检测RNA干扰沉默HPA表达后对SACC-M细胞侵袭和迁移能力的影响。结果:重组质粒pGPH1/GFP/Neo/HPA-siRNA显著降低HPA mRNA和蛋白的表达水平,干扰组中穿透Transwell小室基质的SACC-M细胞数明显低于对照组(P<0.05),划痕损伤实验结果显示,干扰组与对照组细胞迁移距离比较,24、48 h内差异有统计学意义(P<0.05)。结论:沉默人SACC-M细胞中HPA的mRNA和蛋白表达,可抑制SACC-M细胞的侵袭和迁移能力。HPA可能是治疗人类涎腺腺样囊性癌的一个新靶点。Objective：To investigate the role of heparanase（HPA） gene silencing by miRNA interference on invasion and migration of SACC-M cells. Methods：Expression vectors of recombinant plasmids expressing artificial HPA miRNA were constructed and transfected to knockdown HPA expression in SACC-M cells. Expressions of HPA mRNA and protein were detected by RT-PCR and Western blot,respectively. Matrigel invasion assay and scratch-wound assay were performed to examine the invasive and migration ability of the SACC-M cells. Results：Transfection of recombinant plasmid of pGPH1/GFP/Neo/HPA-siRNA significantly suppressed the HPA mRNA and protein levels. This knockdown significantly inhibited the invasion and migration of SACC-M cells as compared with controls. Conclusion： HPA may play an important role in invasion and migration in human salivary adenoid cystic carcinoma cells and should be considered as a potential small molecule inhibitor therapeutic targeting strategy for the future.

关 键 词：腺样囊性癌 基因沉默 乙酰肝素酶 侵袭 转移 

分 类 号：R739.8[医药卫生—肿瘤]