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本文分析了人肝癌细胞株7404,7721细胞中表皮生长因子受体(EGFR)基因表达和EGF对肝癌细胞生长的促进作用。~(125)Ⅰ-EGF对7404细胞的结合试验表明结合是可饱和的和专一的,从~(125)Ⅰ-EGF对7404、7721细胞结合浓度曲线作Scatchard作图和计算,提示每个7404和7721细胞表面分别有1.1×10~5和0.7×10~5的EGFR分子。Northern杂交分析证明EGFR基因在7404,7721细胞中的转录产物主要是5.6 kb EGFR mRNA,免疫印迹分析证明7404细胞和7721细胞的EGFR为170kd的蛋白。EGF对培养于含10%或0.5%小牛血清的RPMI-1640培液中的7404、7721细胞的贴壁依赖性生长有促进作用,促进作用的程度与培液中CS含量有一定关系,提示EGF的促生长作用可能是EGF与血清中其他成分协同作用的结果。EGF对培养于软琼脂中的7404,7721细胞的贴壁不依赖性生长也有明显促进作用。综合上述实验结果说明EGFR基因在人肝癌细胞中是活跃表达的,EGF可能是肝癌细胞生长依赖的一个重要有丝分裂原。
This paper analyzed the expression of epidermal growth factor receptor (EGFR) gene in human hepatocellular carcinoma cell line 7504,7721 cells and the promotion of EGF on hepatoma cell growth. The binding assay of ~(125)I-EGF on 7 440 cells showed that the binding was saturable and specific. Scatchard plots and calculations of ~(125)I-EGF vs. 7404, 7721 cell binding concentration curves suggested that each There are 1.1×10~5 and 0.7×10~5 EGFR molecules on the surface of 7404 and 7721 cells respectively. Northern blot analysis showed that the transcript of EGFR gene in 7404 and 7721 cells was mainly 5.6 kb EGFR mRNA. Immunoblot analysis demonstrated that the EGFR of 7404 cells and 7721 cells was a 170 kd protein. EGF promoted the anchorage-dependent growth of 7404 and 7721 cells cultured in RPMI-1640 culture medium containing 10% or 0.5% calf serum. The degree of the promotion effect was related to the CS content in the culture solution. The growth-promoting effect of EGF may be the result of a synergistic effect of EGF with other components in the serum. EGF also significantly promoted anchorage-independent growth of 7404,7721 cells cultured in soft agar. The above experimental results show that EGFR gene is actively expressed in human hepatoma cells, and EGF may be an important mitogen dependent on the growth of hepatoma cells.