左旋门冬酰胺酶抗白血病作用与胞周Asn水平的关系

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摘要目的：通过检测左旋门冬酰胺酶（L-Asp）处理后培养基中4种氨基酸（Asn、Aspa、Glu和Gln）水平的改变，初步探讨L-Asp的抗白血病作用机制。方法：用CCK-8试剂盒检测L-Asp对 8种常见血液系统恶性肿瘤细胞株（U937，HL-60，Jurkat，NB4，THP-1，Namalwa，Karpass299，K562）的增殖抑制作用 (IC50)；高压液相色谱 (HPLC) 检测培养基中上述4种氨基酸水平的变化。结果：L-Asp在0.01 U/mL作用4 h或1 U/mL作用5 h就可以达到培养基中氨基酸水平的耗竭。但不同血液系统恶性肿瘤细胞对L-Asp的敏感性存在显著差异，最为敏感的细胞株为U937，而Karpass299和K562天然耐药。各种细胞株对L-Asp的敏感性存在明显的剂量效应关系，在较低的药物浓度下，细胞的增殖抑制率处于较低的水平，而药物浓度增加后，细胞的增殖抑制率明显升高。结论：耗竭Asn并非L-Asp发挥作用的有用指标，不同肿瘤对L-Asp的敏感性存在于细胞内部对这种氨基酸缺乏的反应性。L-Asp耗竭Asn只是其发挥作用的前提条件，L-Asp的抗白血病作用与其剂量存在量效关系，因此临床上使用较大剂量L-Asp治疗儿童急性淋巴细胞白血病非常重要。［中国当代儿科杂志，2010，12（7）：557-562］

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	李本尚
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关键词 ：左旋门冬酰胺酶, 白血病, 门冬氨酸, 细胞株

Abstract：OBJECTIVE: To study the antileukemic activity of L-asparaginase through determining the changes of 4 kinds of amino acids (Asn, Aspa, Glu and Gln) in cell culture medium. METHODS: Following L-Asp treatment with designed concentrations and duration, the IC50 (inhibitory concentration 50%) of 8 kinds of common leukemia cell lines (U937, HL-60, Jurkat, NB4, THP-1, Namalwa, Karpass299, K562) were determined by CCK-8 assay. The changes of the 4 kinds of amino acids mentioned above were detected by high performance liquid chromatography (HPLC). RESULTS: The asparagines in cell culture medium were rapidly exhausted when treated with 0.01 U/mL L-Asp for 4 hrs or 1 U/mL L-Asp for 5 minutes. There were significant differences in the sensitivities to L-Asp of different leukemia cell lines. The sensitivities to L-Asp of various cell lines were dose-dependent. Low concentration of L-Asp resulted in a low IC50 and the IC50 increased following the L-Asp concentration increased. CONCLUSIONS: Different leukemia cell lines have different sensitivities to L-Asp, suggesting that exhaustion of asparagines around leukemia cells could not reflect the treatment efficacy of L-Asp. L-Asp antileukemic activity is dose-dependent, which suggests the importance of high-dose L-Asp on childhood acute lymphoblastic leukemia. ［Chin J Contemp Pediatr, 2010, 12 (7):557-562］

Key words： L-asparaginase Leukemia Asn Cell line

PACS:	R733.4
	R730.53

引用本文:

李本尚,罗长缨,何映谊等. 左旋门冬酰胺酶抗白血病作用与胞周Asn水平的关系[J]. 中国当代儿科杂志, 2010, 12(07): 557-562.

LI Ben-Shang,LUO Chang-Ying,HE Ying-Yi et al. Relationship between the antileukemic activity of L-asparaginase and Asn level around leukemic cells[J]. CJCP, 2010, 12(07): 557-562.

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http://www.zgddek.com/CN/ 或 http://www.zgddek.com/CN/Y2010/V12/I07/557

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