ASNA1, an ATPase targeting tail-anchored proteins, regulates melanoma cell growth and sensitivity to cisplatin and arsenite
ASNA1, an ATPase targeting tail-anchored proteins, regulates melanoma cell growth and sensitivity to cisplatin and arsenite
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DOI：
10.1007/s00280-008-0762-2
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发表时间：
2009-02-01
影响因子：
3
通讯作者：
Naredi, P.
中科院分区：
文献类型：
--
作者：
Hemmingsson, O.;Zhang, Y.;Naredi, P.
ASNA1与大肠杆菌ArsA同源，后者是一种具有良好特征的atp酶，参与亚砷酸盐和锑酸盐的外排。对亚砷酸盐和锑酸盐耐药的细胞对化疗药物顺铂有交叉耐药。ASNA1也是将尾锚定蛋白插入内质网膜所必需的atp酶，也是胰岛素分泌的一种新型调节剂。本研究的目的是确定ASNA1水平的改变是否影响人类黑色素瘤细胞对亚砷酸盐和顺铂的生长和敏感性。用含有正义或反义ASNA1的质粒转染培养的黑色素瘤T289细胞。细胞暴露于顺铂、亚砷酸盐和锌中。MTT法和TUNEL法分别评价细胞生长和化学敏感性。ASNA1的表达是生长所必需的。ASNA1表达降低的T289克隆比野生型T289的倍增率高出51 +/- 5% （P = 0.0091）。暴露于顺铂后，ASNA1下调的细胞凋亡显著增加。ASNA1低表达细胞的顺铂IC（50）为野生型的41.7 +/- A 1.8% (P = 0.00097)，亚砷酸盐IC（50）为野生型的59.9 +/- A 3.2% （P = 0.0067）。ASNA1表达降低与细胞生长明显抑制、细胞凋亡增加以及对顺铂和亚砷酸盐敏感性增加有关。
ASNA1 is homologous to E. coli ArsA, a well characterized ATPase involved in efflux of arsenite and antimonite. Cells resistant to arsenite and antimonite are cross-resistant to the chemotherapeutic drug cisplatin. ASNA1 is also an essential ATPase for the insertion of tail-anchored proteins into ER membranes and a novel regulator of insulin secretion. The aim of this study was to determine if altered ASNA1 levels influenced growth and sensitivity to arsenite and cisplatin in human melanoma cells.Cultured melanoma T289 cells were transfected with plasmids containing sense or antisense ASNA1. Cells were exposed to cisplatin, arsenite and zinc. Cell growth and chemosensitivity were evaluated by the MTT assay and apoptosis by a TUNEL assay.ASNA1 expression was necessary for growth. T289 clones with decreased ASNA1 expression exhibited 51 +/- A 5% longer doubling times than wildtype T289 (P = 0.0091). After exposure to cisplatin, ASNA1 downregulated cells displayed a significant increase in apoptosis. The cisplatin IC(50) in ASNA1 underexpressing cells was 41.7 +/- A 1.8% compared to wildtype (P = 0.00097) and the arsenite IC(50) was 59.9 +/- A 3.2% of wildtype IC(50) (P = 0.0067).Reduced ASNA1 expression is associated with significant inhibition of cell growth, increased apoptosis and increased sensitivity to cisplatin and arsenite.