摘要
目的探讨ACP1蛋白对人骨肉瘤细胞侵袭能力的影响。方法以骨肉瘤细胞MG-63为研究对象,针对ACP1基因设计小干扰RNA(si RNA),构建其短发夹状RNA(short hairpin RNA,shRNA)真核表达载体并转染入细胞,在mRNA和蛋白水平检测ACP1表达,通过黏附实验、细胞划痕、transwell检测MG-63体外侵袭能力。结果成功构建si RNA表达载体Pgenesil-1/ACP1-shRNA。shRNA可使ACP1在基因和蛋白表达水平显著降低,骨肉瘤细胞的迁移能力明显下降,黏附能力由(96.4±8.8)%到(43.4±6.0)%,P<0.01;穿过人工基底膜的细胞数量明显减少,由56.5±4.8减少到36.3±6.1,P<0.05。结论ACP1在骨肉瘤的侵袭过程中发挥着重要作用,为肿瘤的生物学治疗提供了新思路。
Objective To study the effects of ACP1 protein on invasion of human osteosarcoma MG-63 cells.Methods A plasmid of a short hairpin RNA（shRNA）targeting ACP1 was constructed and transfected into MG-63 cells.The expression of ACP1 mRNA and protein in MG-63 cells before and after transfection was detected by RT-PCR and Western blot,respectively.The capacity of adhesion,migration and invasion was examined by adhesion assay,wound-healing assay and transwell assay.Results The recombinant plasmid Pgenesil-1/ACP1-shRNA was successfully constructed.shRNA efficiently inhibited the expression of ACP1 mRNA and protein and suppressed cell migration.The adhesion was decreased from（96.4±8.8）% to（43.4±6.0）%（P〈0.01）,and invasion ability from 56.5±4.8 to 36.3±6.1（P〈0.05）.Conclusion Down-regulating ACP1 by shRNA reduced the capacity of proliferation and metastasis of MG-63 cells,which provided a novel approach to bio-therapy of cancer.
出处
《华中医学杂志》
CAS
2009年第4期175-178,共4页
Central China Medical Journal
基金
国家973计划基金(No.2002CB513100)
