NOVEL AND KNOWN PROTEIN-TYROSINE KINASES AND THEIR ABNORMAL EXPRESSION IN HUMAN-MELANOMA
NOVEL AND KNOWN PROTEIN-TYROSINE KINASES AND THEIR ABNORMAL EXPRESSION IN HUMAN-MELANOMA
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DOI：
10.1111/1523-1747.ep12371675
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发表时间：
1993-11-01
影响因子：
6.5
通讯作者：
BENNETT, DC
中科院分区：
文献类型：
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作者：
EASTY, DJ;GANZ, SE;BENNETT, DC
我们已经使用聚合酶链式反应和Northern blotting来鉴定在黑色素瘤的发生和发展过程中可能起重要作用的蛋白酪氨酸激酶。利用酪氨酸激酶基因保守催化结构域的简并引物，从人黑色素瘤细胞系(DX3-LT5.1)和正常人黑素细胞中扩增和克隆了部分c DNA序列。当对黑色素瘤反应产物进行测序时，发现了13个不同的克隆，其中一个是迄今为止的新克隆，暂时被命名为MEK(黑素细胞激酶)。在剩下的12个已知的激酶中，只有两个，ERB-B2和IGF1-R，以前在色素细胞中被报道过。黑素细胞的反应产物只包括这13个序列中的8个。为了测试正常细胞和恶性细胞之间酪氨酸激酶表达的定量差异，用Northern blotting对八个黑色素瘤株和正常黑素细胞进行了分析。两种酪氨酸激酶(JTK-14/Tie和Tyro-9)在一些黑色素瘤中检测到，但在正常黑素细胞中未发现，而包括MEK在内的其他酪氨酸激酶在一些恶性黑色素瘤中似乎过表达。少数蛋白激酶基因表达水平无变化或降低。酪氨酸激酶的表达是独立的，个别品系含有这些酶的不同组合。我们的发现与这些假定的生长因子受体在黑色素瘤发展过程中总体表达增加是一致的。
We have used the polymerase chain reaction and Northern blotting to identify protein tyrosine kinases that may play an important role in the process of melanoma initiation and progression. Degenerate primers from the conserved catalytic domain of tyrosine kinase genes were used to amplify and clone partial cDNA sequences from a human melanoma cell line (DX3-LT5.1) and normal human melanocytes. When the melanoma reaction products were sequenced, 13 distinct clones were found, of Which one is novel to date and has provisionally been named MEK (for melanocytic kinase). Of the remaining 12 known kinases, only two, ERB-B2 and IGF1-R, have previously been reported in pigment cells. Reaction products from melanocytes included only eight of these 13 sequences. To test for quantitative differences in tyrosine kinase expression between normal and malignant cells, a panel of eight melanoma lines and normal melanocytes was analyzed by Northern blotting. Two tyrosine kinases (JTK-14/TIE and TYRO-9) were detected in some melanomas but were not found in normal melanocytes, whereas others, including MEK, appeared to be overexpressed in some malignant lines. A minority of kinases showed either no change or a reduction in the level of mRNA. Expression of tyrosine kinases varied independently, and individual lines contained various combinations of these enzymes. Our findings are consistent with an increased overall expression of these putative growth factor receptors during melanoma development.