尿激酶受体反义RNA抑制人乳腺癌细胞的侵袭作用

将尿激酶受体 u PAR反义 RNA表达质粒 p URAS以脂质体法转染高侵袭性人乳腺癌细胞株 MDA- MB- 2 31 ,G41 8筛选抗性克隆 .Northern印迹法检测 u PAR反义 RNA的表达 ,RT- PCR法检测 u PAR的表达 ,牛奶板法测定细胞培养上清中纤溶活性 .改良 Boyden小室模型和裸小鼠乳房脂肪垫接种试验分别检测肿瘤细胞体外和体内侵袭能力 .反义克隆细胞能表达 u PAR反义RNA,其 u PAR表达水平及培养上清中纤溶活性明显降低 .反义细胞克隆体外侵袭能力比原代细胞 MDA- MB- 2 31和转染载体细胞克隆显著降低 .裸小鼠体内侵袭实验表明 ,反义细胞克隆的成瘤性、生长性和侵袭性均显著受到抑制 .u PAR至少在一部分恶性乳腺癌侵袭行为中发挥重要作用 ,反义 RNA可望成为抗肿瘤侵袭治疗的一种有效手段 .

Inhibition of Invasiveness of Human Breast Cancer Cells by Antisense RNA for Urokinase Receptor

Urokinase receptor antisense RNA expression plasmid,pURAS was introduced into the highly invasive human breast cancer cell line MDA MB 231 with lipofectin,and positive clones were screened out with G418.Northern blot method was applied to detect the expression of antisense RNA,and RT PCR method was employed to detect the expression of uPAR.The plasminogen activity of culture supernatant was detected with milk plate method.The in vitro and in vivo invasive ability of cancer cells was defined with Boyden's chamber model and nude mice transplantation,respectively.One of the clones MAS4 was found to express antisense RNA for uPAR.Its expression of uPAR and plasminogen activity in culture supernatant were significantly decreased.The in vitro invasive ability of antisense clone was significantly compromised in comparison with both parental cell and pcDNA3 vector\|only transfected cell.The in vivo invasion experiment on mice showed the tumorigenicity,growth and invasiveness of MAS4 had been significantly inhibited.Antisense uPAR significantly decreased the invasiveness both in vitro and in vivo ,as well as the tumorigenicity and growth of at least a certain kind of human breast cancer.The antisense technique is expected to become one effective method in anti\|invasion therapy of cancer.