双调蛋白诱导乳腺癌细胞表达尿激酶型纤溶酶原活化物

目的:在人乳腺癌模型上研究双调蛋白与尿激酶型纤溶酶原活化物表达之间的关系.方法:乳腺癌NS2T2A1细胞经双调蛋白反义cDNA质粒转染后经潮霉素B筛选获得表达双调蛋白反义RNA的AR-AS1及AR-AS3两个细胞克隆,转染空载体获得NS2T2A1 V对照细胞,接种至裸鼠皮下形成肿瘤.测定细胞及肿瘤uPA表达水平,并研究uPA与细胞侵袭性之间的关系.结果:AR-AS1及AR-AS3细胞体外及体内uPA表达均被抑制.外源性双调蛋白可刺激对照细胞uPA的表达,并部分恢复AR-AS1及AR-AS3细胞uPA的表达水平.双调蛋白反义cDNA质粒转染及抗uPA抗体均导致乳腺癌细胞体外侵袭性的降低.结论:在乳腺癌模型上,uPA表达与肿瘤细胞的侵袭密切相关.双调蛋白反义RNA表达可有效地抑制uPA的表达,进而抑制肿瘤细胞的侵袭性.

Amphiregulin Induces uPA Expression in Breast Cancer Cells

Objective:To investigate the relationship between amphiregulin expression and uPA production in a human breast cancer model.Methods:Human amphiregulin cDNA antisense plasmid was transfected in NS2T2A1 cells.Two clones,selected by hygromycin B,expressed AR antisense RNA(AR AS1 and AR AS3 cell lines)in which AR protein expression was reduced.Control cell line NS2T2A1 V was obtained by empty vector transfection.These cells were injected subcutaneously in nude mice to obtain tumors.The in vitro and in vivo uPA production were detected,and the correlation between uPA expression and invasive capability of tumor cells was studied.Results:The in vitro and in vivo uPA expression were inhibited in AR AS1 and AR AS3 cells in comparison with that of the control cells.Exogenous amphiregulin stimulated uPA production in control cells and partially restored the uPA level in AS1 and AR AS3 cells.Amphiregulin cDNA antisense transfection as well as anti-uPA antibody treatment suppressed the invasive capability of tumor cell.Conclusion:The uPA expression is closely related with tumor cell invasion in the breast cancer model.Amphiregulin antisense RNA expression can efficiently inhibit uPA production and the invasive capability of tumor cell.