Tsai L H, Nanu L, Smith R G, Ozanne B
CRC Laboratories, Beatson Institute for Cancer Research, Glasgow, UK.
Oncogene. 1991 Jan;6(1):81-8.
The c-fos proto-oncogene is found to be overexpressed at least 30-fold in SMS-SB, a pre-B leukemic cell line compared to other cell types. No gross alteration of the c-fos gene structure in SMS-SB cells can be detected by karyotypic or Southern analyses. C-fos in SMS-SB cells can still be induced by serum, TPA and the calcium ionophore, A23187, and superinduced by the combination of serum and cycloheximide. The elevated levels of c-fos transcripts are not due merely to an increased life span of mRNA, as the half-life of steady state c-fos mRNA in SMS-SB cells is about 40 min. Nuclear run-on transcription assays demonstrate that the transcription rate of c-fos in SMS-SB cells is up-regulated about 30-fold as compared to another pre-B leukemic cell line, NALM-6. In order to determine whether this is a cis- or transacting effect, we sequenced the 550 base pairs upstream of the SMS-SB c-fos coding region and found no mutations upstream of the mRNA CAP site. However, two point mutations at positions +23 and +98, respectively, were found in the 5' noncoding region of the first exon. Consistent with the overexpression of c-fos mRNA, the 55 kD c-fos protein is present in SMS-SB cells through detection by immunoprecipitation, but could not be detected in NALM-6 cells. SMS-SB cells however, do not appear to contain more abundant AP-1 DNA binding activity than NALM-6 cells.
与其他细胞类型相比,在一种前B淋巴细胞白血病细胞系SMS-SB中发现原癌基因c-fos的表达至少上调了30倍。通过核型分析或Southern分析未检测到SMS-SB细胞中c-fos基因结构的明显改变。SMS-SB细胞中的c-fos仍可被血清、佛波酯(TPA)和钙离子载体A23187诱导,并被血清与环己酰亚胺联合作用超诱导。c-fos转录本水平的升高不仅仅是由于mRNA寿命的延长,因为SMS-SB细胞中稳态c-fos mRNA的半衰期约为40分钟。核转录分析表明,与另一种前B淋巴细胞白血病细胞系NALM-6相比,SMS-SB细胞中c-fos的转录速率上调了约30倍。为了确定这是顺式作用还是反式作用,我们对SMS-SB c-fos编码区上游550个碱基对进行了测序,发现在mRNA帽位点上游没有突变。然而,在第一个外显子的5'非编码区分别发现了位于+23和+98位置的两个点突变。与c-fos mRNA的过表达一致,通过免疫沉淀检测发现55 kD的c-fos蛋白存在于SMS-SB细胞中,但在NALM-6细胞中未检测到。然而,SMS-SB细胞似乎并不比NALM-6细胞含有更丰富的AP-1 DNA结合活性。