Activation of protein kinase C-alpha isoform in murine melanoma cells with high metastatic potential
Activation of protein kinase C-alpha isoform in murine melanoma cells with high metastatic potential
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DOI：
10.1023/a:1018447531813
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发表时间：
1997-11-01
影响因子：
4
通讯作者：
Comolli, R
中科院分区：
文献类型：
--
作者：
LaPorta, CAM;Comolli, R
转移是一个多步骤的过程，其中蛋白激酶C（PKC）似乎显着参与。我们分析了体外不同培养条件下B16-F1和B16-BL 6黑色素瘤细胞中经典（α，β，γ）和新型PKC β亚型的活性和表达。我们在不同的培养基（分别为DMEM或RPMI 1640）中使用高浓度和低浓度的酪氨酸和苯丙氨酸，其影响细胞的转移潜能和增殖能力。我们还测试了弱转移性无黑色素B78-H1黑色素瘤细胞系，其不受不同培养条件的影响。在两种B16黑色素瘤细胞系中，PKC α的活化（表达没有增加）发生在允许转移的生长条件（DMEM）下。相反，弱转移性无黑色素B78-H1细胞系在两种不同的培养基中表现出该亚型的实质性失活，表明PKC α特异性参与转移过程。此外，在B16黑色素瘤细胞中，新的PKC α在刺激生长但不刺激转移的培养条件下被激活（RPMI 1640）。为了确定PKC激活和转移过程之间的关系，我们还测定了组织蛋白酶B的释放。PKC活性和组织蛋白酶B释放之间没有相关性，在B16黑色素瘤细胞系可以证明。
Metastasis is a multistep process in which protein kinase C (PKC) appears to be significantly involved. We analysed the activity and expression of classical (alpha, beta, gamma) and novel PKC epsilon isoforms in B16-F1 and B16-BL6 melanoma cells maintained under different culture conditions in vitro. We used high and low concentrations of tyrosine and phenylalanine in different media (DMEM or RPMI 1640 respectively) that affect the metastatic potential and also the proliferative, capacity of the cells. We also tested a weakly metastatic amelanotic B78-H1 melanoma cell line which is unaffected by the different culture conditions. In both B16 melanoma tell lines activation of PKC alpha (without increased expression) occurred under growth conditions permissive of metastasis (DMEM). In contrast, the weakly metastatic amelanotic B78-H1 cell line showed a substantial inactivation of this isoform in the two different culture media, suggesting a specific involvement of PKC alpha in the metastatic process, Moreover, in B16 melanoma cells, novel PKC epsilon was activated under culture conditions which stimulated grow th but not metastasis (RPMI 1640). In order to define the relationship between PKC activation and the metastatic process we also determined the release of cathepsin B. No correlation between PKC activity and cathepsin B release in either B16 melanoma cell lines could be demonstrated.