ANALYSIS OF ABNORMAL EXPRESSION OF G-CSF GENE IN A NOVEL TUMOR-CELL LINE (KHC-287) ELABORATING G-CSF, IL-1 AND IL-6 WITH CO-AMPLIFICATION OF C-MYC AND C-KI-RAS
ANALYSIS OF ABNORMAL EXPRESSION OF G-CSF GENE IN A NOVEL TUMOR-CELL LINE (KHC-287) ELABORATING G-CSF, IL-1 AND IL-6 WITH CO-AMPLIFICATION OF C-MYC AND C-KI-RAS
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DOI：
10.1002/ijc.2910480321
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发表时间：
1991-05-30
影响因子：
6.4
通讯作者：
IMURA, H
中科院分区：
文献类型：
--
作者：
SUZUKI, A;TAKAHASHI, T;IMURA, H
我们从一名伴有明显白细胞增多的大细胞型肺癌患者中建立了人类癌细胞系 (KHC 287)。 KHC 287细胞的培养上清液促进半固体培养中人骨髓细胞的粒细胞集落形成。用单克隆抗粒细胞集落刺激因子 (G-CSF) 抗体处理上清液，集落形成几乎完全被抑制。通过ELISA法，在培养上清液中不仅检测到G-CSF，还检测到白细胞介素-1-α(IL-1-α)、IL-1-β和IL-6。 KHC 287 细胞的 Northern 印迹分析揭示了这些细胞因子基因的独特表达。 KHC 287 DNA 的 Southern 印迹杂交显示 c-myc 和 c-ki-ras 分别共扩增 20 倍和 40 倍。在用插入 pSV00CAT 的 G-CSF 基因的 360 bp 上游区域转染的 KHC 287 细胞中，氯霉素乙酰转移酶 (CAT) 活性明显增强，但在不产生 G-CSF 的肿瘤细胞系中则没有。这些结果表明，与G-CSF上游区域360bp结合的反式激活因子的过量产生是KHC 287细胞系中G-CSF基因异常表达的原因。
We established a human carcinoma cell line (KHC 287) from a patient wiht large-cell-type lung carcinoma associated with marked leukocytosis. The culture supernantant of KHC 287 cells promoted granulocytic colony formation of human bone-marrow cells in semi-solid culture. Colony formation was almost completely suppressed by treatment of the supernatant with a monoclonal anti-granulocyte colony-stimulating factor (G-CSF) antibody. Not only G-CSF but also interleukin-l-alpha (IL-l-alpha), IL-l-beta and IL-6 were detected in the culture supernatant by an ELISA method. Northern blot analysis of KHC 287 cells revealed distinct expression of these cytokine genes. Southern blot hybridization of KHC 287 DNA showed 20- and 40-fold co-amplification of c-myc and c-ki-ras, respectively. The chloramphenicol acetyl transferase (CAT) activity was distinctly enhanced in the KHC 287 cells which were transfected with the 360 bp upstream region of G-CSF gene inserted into pSV00CAT, but not in non-G-CSF-producing tumor cell lines. These results suggest that over-production of the transactivating factor(s) which binds to the 360 bp of the G-CSF upstream region is responsible for the abnormal expression of G-CSF gene in KHC 287 cell line.