Reversal of the transformed phenotype and inhibition of peptidylglycine alpha-monooxygenase in Ras-transformed cells by 4-phenyl-3-butenoic acid.
Reversal of the transformed phenotype and inhibition of peptidylglycine alpha-monooxygenase in Ras-transformed cells by 4-phenyl-3-butenoic acid.
复制标题
4-苯基-3-丁烯酸逆转 Ras 转化细胞中的转化表型并抑制肽基甘氨酸 α-单加氧酶。
DOI：
10.1002/mc.20060
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发表时间：
2004
期刊：
影响因子：
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通讯作者：
Matesic,DianeF
中科院分区：
文献类型：
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作者：
Sunman,JeffreyA;Foster,MichaelS;Folse,StaceyL;May,SheldonW;Matesic,DianeF
最近的研究表明，一些肿瘤细胞的增殖依赖于需要酰胺化自分泌生长因子的自分泌生长环。肽基甘氨酸α-单加氧酶（PAM）是这些生长因子酰胺化所必需的，因此，这种酶是抗肿瘤化合物的一个有吸引力的靶标。4-苯基-3-丁烯酸（PBA）是一种体外不可逆的PAM周转依赖性抑制剂，已被证明可通过抑制酰胺化生长因子的合成来降低肺癌细胞增殖。我们发现PBA（0.1 mg/mL）抑制Ras转化上皮细胞（WB‐Ras）的生长，但对正常上皮细胞（WB‐Neo）的增殖几乎没有影响。与正常上皮细胞生长相比，浓度低10倍的PBA甲酯衍生物（PBA-Me）也显示出对Ras转化细胞生长的选择性抑制。此外，PBA处理2-5天后，WB‐ Ras细胞之间的间隙连接通讯显著上调，连接蛋白43磷酸化程度相应增加，含连接蛋白43的质膜间隙连接斑块数量增加。Western印迹分析表明PBA对细胞膜与胞浆中p21 Ras的比例或对p44/42 MAP激酶磷酸化没有影响。此外，PBA处理的WB-Ras细胞的细胞形态发生改变，从而更接近于未转化的WB-Neocell的细胞形态。在WB-Ras和WB-Neocell中测定PAM活性，并且我们证明PBA在长处理时间（4天）下在产生WB-Ras细胞的选择性生长抑制的浓度下抑制两种细胞类型中的PAM活性。然而，较短的PBA处理时间（24小时）抑制WB-Ras中的PAM活性，但不抑制WB-新细胞中的PAM活性，这是PBA-Me模拟的效果。综上所述，这些结果清楚地表明PBA使Ras转化的细胞恢复为更正常的表型，这一发现与转化的上皮细胞中已知的Rassignaling途径的优势增加一致。© 2004 Wiley利斯公司
Recent studies have shown that the proliferation of some tumor cells is dependent on autocrine growth loops that require amidated autocrine growth factors. Peptidylglycine α‐monooxygenase (PAM) is required for amidation of these growth factors and, therefore, this enzyme is an attractive target for anti‐tumor compounds. 4‐Phenyl‐3‐butenoic acid (PBA) is an irreversible turnover‐dependent inhibitor of PAM in vitro and has been shown to decrease lung cancer cell proliferation by inhibiting the synthesis of amidated growth factors. We show here that PBA (0.1 mg/mL) inhibits the growth ofRas‐transformed epithelial cells (WB‐Ras) but has little effect on the proliferation of normal epithelial cells (WB‐Neo). The methyl ester derivative of PBA (PBA‐Me) at 10‐fold lower concentration also exhibits a selective inhibition ofRas‐transformed cell growth compared to normal epithelial cell growth. In addition, PBA produces a significant upregulation of gap junctional communication between WB‐Rascells following 2–5 day treatments, with a corresponding increase in the degree of connexin 43 phosphorylation and an increase in the number of connexin 43‐containing plasma membrane gap junction plaques. Western blot analyses indicate no effect of PBA on the proportion of p21Rasin the membrane versus cytosolic fractions or on p44/42 MAP kinase phosphorylation. Furthermore, the cell morphology of PBA‐treated WB‐Rascells is altered, so as to more closely resemble that of non‐transformed WB‐Neocells. PAM activity was assayed in both WB‐Rasand WB‐Neocells, and we demonstrate that PBA at long treatment times (4 days) inhibits PAM activity in both cell types at concentrations that produce selective growth inhibition of WB‐Rascells. Shorter PBA treatment times (24 h), however, inhibit PAM activity in WB‐Rasbut not WB‐Neocells, an effect that was mimicked by PBA‐Me. Taken together, these results clearly demonstrate that PBA returnsRas‐transformed cells to a more normal phenotype, a finding consistent with the known increased dominance of theRassignaling pathway in transformed epithelial cells. © 2004 Wiley‐Liss, Inc.