Yamada T, Odawara K, Kaneko H
Environmental Health Science Laboratory, Sumitomo Chemical Co. Ltd., 1-98, 3-Chome, Kasugade-naka, Konohana-ku, Osaka 554-8558, Japan.
Mutat Res. 2000 Nov 20;471(1-2):29-36. doi: 10.1016/s1383-5718(00)00115-7.
We previously established a transgenic Chinese hamster CHL/IU cell line, designated as KN63, for concurrent analysis of gene mutations and chromosome aberrations. The KN63 cell line contains copies of a shuttle vector with the Escherichia coli gpt gene as a mutational target in its chromosome. To evaluate the sensitivity of the cell line to various types of mutagens, methyl methanesulfonate (MMS), N-ethyl-N-nitrosourea (ENU), mitomycin C (MMC), vincristine sulfate (VIN) and C.I. basic red 9 hydrochloride (CIB) were assayed. KN63 cells were treated with each test chemical and gene mutations were detected in the gpt gene of the shuttle vector rescued from the KN63 cell genome into an E. coli host. Chromosome aberrations were concurrently evaluated by conventional metaphase analysis. MMS, ENU and MMC induced both gene mutations and structural chromosome aberrations in KN63 cells, with more efficient induction of the latter. VIN, a well-known aneugen, produced only numerical changes to chromosomes, while CIB was negative for both types of alteration. KN63 cells were as sensitive to MMS, ENU, MMC and VIN as Chinese hamster cell lines such as CHL, CHO and V79 cells. The characteristics of test chemicals indicated by this system should be useful for understanding endpoints in chemical mutagenesis.
我们之前建立了一种转基因中国仓鼠CHL/IU细胞系，命名为KN63，用于同时分析基因突变和染色体畸变。KN63细胞系在其染色体中含有携带大肠杆菌gpt基因作为突变靶点的穿梭载体拷贝。为了评估该细胞系对各种类型诱变剂的敏感性，对甲磺酸甲酯（MMS）、N-乙基-N-亚硝基脲（ENU）、丝裂霉素C（MMC）、硫酸长春新碱（VIN）和盐酸碱性品红9（CIB）进行了检测。用每种测试化学品处理KN63细胞，并在从KN63细胞基因组拯救到大肠杆菌宿主中的穿梭载体的gpt基因中检测基因突变。同时通过常规中期分析评估染色体畸变。MMS、ENU和MMC在KN63细胞中诱导了基因突变和结构性染色体畸变，对后者的诱导更有效。VIN是一种众所周知的非整倍体诱变剂，仅引起染色体数目变化，而CIB对两种类型的改变均为阴性。KN63细胞对MMS、ENU、MMC和VIN的敏感性与中国仓鼠细胞系如CHL、CHO和V79细胞相同。该系统所显示的测试化学品的特性应有助于理解化学诱变中的终点。
