Li Ben-Shang, He Ying-Yi, Luo Chang-Ying, Jiang Hua, Shen Shu-Hong, Jiang Li-Min, Zhang Bei, Gu Long-Jun
Department of Hematology/Oncology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Jun;18(3):559-63.
This study was purposed to explore the relationship between asparagine synthetase (AsnS) mRNA expression level and the sensitivity of leukemic cell lines to L-asparaginase. The AsnS mRNA expression level in 8 cell lines (Jurkat, HL-60, U937, NB4, THP-1, Namalwa, Karpas299 and K562) was determined by real-time quantitative PCR (RQ-PCR) based on fluorescence dye Eva Green before and after treatment with L-Asp, and the cell proliferation rates were analyzed by CCK-8 assay. The results showed that there was a significant disparity of AsnS expression level in 8 cell lines, and there were significant increases of AsnS expression level in cells co-cultured with L-Asp (p < 0.05). Of all these eight cell lines, cells sensitive to L-asparaginase had lower AsnS expression level and cells resistant to L-asparaginase had higher AsnS expression. U937 which was the most sensitive to L-asparaginase had the lowest AsnS expression level, while K562 was natural resistant to L-asparaginase and possessed of the highest AsnS level. It is concluded that the AsnS plays a critical role in regulating cellular biological behavior after depletion of asparagine, the AsnS mRNA expression level in cells reflects the sensitivity of cells to L-Asp. The results may imply the possibility for the use of L-asparaginase in leukemia with lower AsnS expression level.
本研究旨在探讨天冬酰胺合成酶(AsnS)mRNA表达水平与白血病细胞系对L-天冬酰胺酶敏感性之间的关系。采用基于荧光染料Eva Green的实时定量PCR(RQ-PCR)法,检测8种细胞系(Jurkat、HL-60、U937、NB4、THP-1、Namalwa、Karpas299和K562)在L-天冬酰胺(L-Asp)处理前后的AsnS mRNA表达水平,并通过CCK-8法分析细胞增殖率。结果显示,8种细胞系的AsnS表达水平存在显著差异,与L-Asp共培养的细胞中AsnS表达水平显著升高(p<0.05)。在这8种细胞系中,对L-天冬酰胺酶敏感的细胞AsnS表达水平较低,而对L-天冬酰胺酶耐药的细胞AsnS表达水平较高。对L-天冬酰胺酶最敏感的U937细胞AsnS表达水平最低,而K562细胞对L-天冬酰胺酶天然耐药且AsnS水平最高。结论:AsnS在天冬酰胺耗竭后调节细胞生物学行为中起关键作用,细胞中AsnS mRNA表达水平反映细胞对L-Asp的敏感性。该结果可能提示L-天冬酰胺酶用于AsnS表达水平较低的白血病治疗的可能性。