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目的 :验证所构建的组织型纤溶酶原激活剂 (tPA)乳腺特异性表达载体pWTA在乳癌细胞系MCF 7中的瞬时表达。方法 :用脂质转染胺试剂LipofectamineTM2 0 0 0 将pWTADNA转染入乳癌细胞系MCF 7和食管癌细胞系Eca 1 0 9。应用原位杂交技术 ,以地高辛标记的tPAcDNA为探针 ,观察PWTA的瞬时表达。结果 :不同浓度LipofectamineTM2 0 0 0MCF 7细胞原位杂交均为阳性 ;Eca 1 0 9细胞及未加转染液者未见阳性信号。结论 :载体pWTA为tPA乳腺特异性表达载体。
Objective : To verify the transient expression of the constructed tissue-type plasminogen activator (tPA) breast-specific expression vector pWTA in breast cancer cell line MCF 7. METHODS: pWTA DNA was transfected into breast cancer cell line MCF 7 and esophageal cancer cell line Eca 109 using lipofectamine 2000. Using in situ hybridization technique, digoxigenin-labeled tPA cDNA was used as a probe to observe the transient expression of PWTA. Results : The in situ hybridization was positive in different concentrations of LipofectamineTM200MCF 7 cells. There was no positive signal in Eca109 cells and those without transfection. Conclusion : Vector pWTA is tPA mammary gland-specific expression vector.