摘要
本文将构建的活化癌基因Ｔ２４－ｒａｓｃＤＮＡ的正、反义真核表达重组体，按先正义后反义和先反义后正义的不同转染顺序，转染ＮＩＨ３Ｔ３细胞，经药物筛选，建立细胞系。通过细胞形态及生物学特性观察、Ｓｏｕｔｈｅｒｎ杂交实验、软琼脂集落形成实验和Ｐ２１蛋白表达量的检测等实验对转化细胞进行了研究。结果表明：本室构建的正义重组体能诱导ＮＩＨ３Ｔ３细胞转化，而反义重组体可有效抑制这一转化，并且抑制效果与正、反义质粒转染的先后顺序有关。
In this paper,the sense and antisense mammalian expression vectors of T24-ras activated oncogene were introduced into cultured NIH3T3 cells in sens→antisense and antisense→sense orders.The transfected cells were selected by drug selection.The morphology change,Southern hybridization,frequency of soft agar colonies and the detection of P21 amount were investigated.The results showed that the sense recombinant could cause the malignant transformation of NIH3T3 cells while the antisense recombinant could inhibite this kind of transformation.Furthermore,the inhibitory effect varied according to the different transfected orders.
关键词
肿瘤
癌基因
反义RNA
Oncogene,DNA transfection,Antisense RNA
