Asparagine synthetase is partially localized to the plasma membrane and upregulated by L-asparaginase in U937 cells
Asparagine synthetase is partially localized to the plasma membrane and upregulated by L-asparaginase in U937 cells
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U937 细胞中天冬酰胺合成酶部分定位于质膜并被 L-天冬酰胺酶上调
DOI：
10.1007/s11596-011-0243-4
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Longjun Gu
中科院分区：
文献类型：
--
作者：
Yingyi He;Ben;C. Luo;S. Shen;J. Chen;H. Xue;Jingyan Tang;Longjun Gu
本研究探讨了天冬酰胺合成酶（ASNS）的细胞内定位与白血病化疗耐药的关系。将pIRES-GFP-ASNS-Flag/Neo表达载体分别瞬时转染SK-N-MC细胞和297 T细胞。免疫荧光和Western blot分析ASNS的细胞定位。用L-天冬酰胺酶处理U937细胞48 h，通过免疫荧光染色检测质膜上内源性ASNS的表达。免疫荧光染色显示ASNS部分定位于转染的SK-N-MC细胞表面。Western blotting结果显示ASNS在-293T细胞的胞浆和质膜上均有表达。用抗ASNS特异性单克隆抗体免疫荧光染色显示，内源性ASNS除了分布在胞液中外，还定位于U937细胞质膜上。此外，ASNS在细胞膜上的表达量在L-天冬酰胺酶处理后明显高于未处理细胞。结论：ASNS的亚细胞易位可能在白血病细胞L-天冬酰胺酶耐药中起重要作用。
SummaryThis study investigated the intracellular localization of asparagine synthetase (ASNS) in the relation with chemoresistance in leukemia. pIRES-GFP-ASNS-Flag/Neo expression vector was transiently tansfected into SK-N-MC cells and 297T cells respectively. Immunofluorescence and Western blot analysis were performed for cellular localization of ASNS respectively. U937 cells were treated with L-asparaginase for 48 h and examined for endogenous ASNS expression on plasma membrane by immunofluorescence staining. Immunofluorescence staining showed that the transiently expressed ASNS was partly localized on transfected-SK-N-MC cell surface. Moreover, Western blotting exhibited that ASNS expressed both in cytosol and on plasma membrane of transfected-293T cells. Immunofluorescence staining with anti-ASNS-specific monoclonal antibody revealed that endogenous ASNS was localized on the plasma membrane of U937 cells, except for its distribution in the cytosol. In addition, ASNS exhibited a higher expression on plasma membrane after treatment with L-asparaginase as compared with the untreated cells. It was concluded that the subcellular translocation of ASNS may play an important role in L-asparaginase resistance in leukemia cells.
DOI：
10.1002/9780470123188.ch5
发表时间：
1998
期刊：
Advances in enzymology and related areas of molecular biology
影响因子：
--
作者：
Nigel G. J. Richards;S. Schuster
通讯作者：
Nigel G. J. Richards;S. Schuster
影响因子：
4.1
作者：
Aslanian, AM;Kilberg, MS
通讯作者：
Kilberg, MS
DOI：
10.1152/ajpcell.1997.272.5.c1691
发表时间：
1997-05-01
影响因子：
5.5
作者：
Hutson, RG;Kitoh, T;Kilberg, MS
通讯作者：
Kilberg, MS