权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Expression of cellular oncogenes in human salivary gland adenocarcinoma cell line

细胞癌基因在人唾液腺腺癌细胞系中的表达

基本信息

批准号：
06671868
负责人：
SATO Nobuko
金额：
$ 1.34万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671868/
关键词：
Salivary gland adenocarcinoma Celluar oncogene EGF EGF receptor Tyrosine kinase MAP kinase c-fos c-myc

项目摘要

The effect of EGF on the cell growth and EGF receptor : The effect of EGF on the cell growth and EGF receptor was examined in HSG-AZA 3 cells. When the cells were treated with EGF,DNA synthesis was approximately 3-fold higher than that without EGF treatment. Western blotting using anti-human EGF receptor antibody showed the band around 170 kDa estimated as EGF receptor, and EGF showed the more high density band with 170 kDa as compared to that without EGF.EGF receptor mRNA was analyzed by RT-PCR using specific primers for EGF receptor. EGF receptor cDNA with 142 bp in unstimulated cells was observed and a maximal enhancement of this 142 bp band was obtained 12 hr after EGF stimulation.The effect of EGF on activities of EGF receptor-associated tyrosine kinase and MAP kinase : Autophosphorylation of EGF receptor was analyzed by Western blotting using a specific antibody against phosphotyrosine. Although there was no phosphorylated band in the region corresponding to EGF receptor in unstimulated cells, a phosphorylated tyrosine band appeared around 170 kDa 5 min after EGF stimulation. Furthermore, the activity of EGF receptor-associated tyrosine kinase was enhanced after EGF treatment using the specific substrate for tyrosine kinase. On the other hand, MAP kinase phosphorylated after EGF stimulation was detected by a specific antibody against phospho-MAP kinase. In addition, the activity of MAP kinase increased 15 min after EGF stimulation.Analysis of c-fos and c-myc proto-oncogenes : Cellular oncogenes, c-myc, c-jun or c-fos which are induced after simulation of growth factors, may play a role as nuclear transcriptional factors. By Western blotting, the band around 62 kDa which seems to be c-fos was found. The intensity of this band increased during 3-6 hr treatment of EGF and then gradually decreased. The effect of EGF on c-fos mRNA and c-myc mRNA was also examined by RT-PCR.EGF caused maximal enhancement of c-fos mRNA at 30 min and that of c-myc mRNA at 2 hr.

EGF对细胞生长和EGF受体的影响：在HSG-AZA 3细胞中检测EGF对细胞生长和EGF受体的影响。当细胞用EGF处理时，DNA合成比没有EGF处理的细胞高约3倍。用抗人EGF受体抗体进行的Western blotting显示了170 kDa左右的EGF受体条带，EGF在170 kDa附近显示了比不含EGF的EGF更高的密度条带。EGF受体cDNA在未刺激的细胞中为142 bp，在EGF刺激后12小时，这条142 bp的条带得到最大增强，EGF对EGF受体相关酪氨酸激酶和MAP激酶活性的影响：用磷酸酪氨酸特异性抗体通过Western印迹分析EGF受体的自磷酸化。虽然没有磷酸化带在相应的区域EGF受体在未刺激的细胞，磷酸化酪氨酸带出现约170 kDa的EGF刺激后5分钟。此外，EGF受体相关的酪氨酸激酶的活性增强EGF治疗后，使用酪氨酸激酶的特异性底物。另一方面，MAP激酶磷酸化EGF刺激后检测到的磷酸化MAP激酶的特异性抗体。c-fos和c-myc原癌基因分析：c-myc、c-jun或c-fos等细胞癌基因在生长因子刺激下被诱导，可能作为核转录因子发挥作用。Western blotting结果显示，EGF作用于细胞后，在62 kDa处出现一条类似c-fos的条带，该条带强度在EGF作用3-6 hr时逐渐增强，随后逐渐减弱。用RT-PCR检测EGF对c-fos mRNA和c-myc mRNA的影响，EGF使c-fos mRNA在30 min和c-myc mRNA在2 h时表达增强最明显。