Yamada H, Koizumi S, Kimura M, Shimizu N
National Institute of Industrial Health, Kawasaki, Japan.
Exp Cell Res. 1989 Sep;184(1):90-8. doi: 10.1016/0014-4827(89)90367-4.
An expression vector was constructed from part of pSV2neo with the 3'-ClaI fragment of the epidermal growth factor (EGF) receptor cDNA inserted in an inverted orientation downstream from the human metallothionein (MT) IIa promoter. The human squamous carcinoma cell line NA, which overproduces EGF receptor, was transfected with this vector and selected for resistance to the neomycin derivative G418. One of the stable transfectants had a 90% reduction in cell-surface EGF receptor in response to ZnSO4. The nascent EGF receptor peptide was also decreased with concurrent induction of MT mRNA. These data suggest that the antisense transcript regulated by the MT promoter inhibits the expression of the endogenous EGF receptor genes. Although no transcripts from the antisense gene were detected, the results indicate that transfection with the antisense vector provides a technique by which to modulate the number of EGF receptors on the cell surface of squamous cell carcinomas.
从pSV2neo的一部分构建了一个表达载体，将表皮生长因子（EGF）受体cDNA的3'-ClaI片段以反向方向插入人金属硫蛋白（MT）IIa启动子下游。过度产生EGF受体的人鳞状癌细胞系NA用该载体转染，并选择对新霉素衍生物G418具有抗性的细胞。其中一个稳定转染子在加入ZnSO4后细胞表面EGF受体减少了90%。同时MT mRNA的诱导伴随着新生EGF受体肽的减少。这些数据表明，由MT启动子调控的反义转录物抑制内源性EGF受体基因的表达。虽然未检测到反义基因的转录物，但结果表明用反义载体转染提供了一种调节鳞状细胞癌细胞表面EGF受体数量的技术。
