Zhang Jin-Yan, Zhang Ju-Shun, Xu Zhen-Shu
Fujian Institute of Hematology, Fujian Provincial Key Laboratory of Hematology, Fujian Medical University Union Hospital, Fuzhou 350000, Fujian province, China.
Fujian Institute of Hematology, Fujian Provincial Key Laboratory of Hematology, Fujian Medical University Union Hospital, Fuzhou 350000, Fujian province, China . E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Aug;23(4):919-24. doi: 10.7534/j.issn.1009-2137.2015.04.002.
To investigate the expression of Notch gene in chronic lymphocytic leukemia cells and to explore the change of Notch protein after the therapy with cytosine arabinoside or dexmethasone, and the mechanism of Notch mediated anti-apoptosis and drug-resistance in chronic lymphocytic leukemia cells.
The mononuclear cells from bone marrow or peripheral blood of chronic lymphocytic leukemia patients (24 cases) and healthy donors (14 cases) were collected, then the expression of Notch gene, BCL-2, as well as NF-κB gene were detected by real-time fluorescent quantitative PCR (qRT-PCR) at the level of transcription. The change of Notch protein in L1210 cell lines after therapy with cytosine arabinoside and dexmethasone was determined by Western blot.
mRNA expression levels of Notch1, Notch2, BCL-2 and NF-κB gene in CLL group were significantly higher than those in healthy control group (0.8556 ± 0.8726 vs 0.6731 ± 0.5334, P = 0.0182; 1.2273 ± 0.8207 vs 0.6577 ± 0.6424, P < 0.0001; 8.0960 ± 7.5661 vs 0.5969 ± 0.4976, P < 0.0001; 1.0966 ± 0.6925 vs 0.5373 ± 0.7180, P < 0.0001, respectively), but no significant difference was found between Notch3 and Notch4 gene (1.1914 ± 2.4219 vs 0.8713 ± 0.7937, P = 0.3427; 0.8174 ± 1.0869 vs 0.9752 ± 1.3446, P = 0.2402, respectively). Notch1 protein expression in L1210 cells were significantly decreased after treating with cytosine arabinoside of low and middle concentrations, but increased after treating with cytosine arabinoside of high concentration or prolonging time of cytosine arabinoside of middle con-centration. Notch1 protein expression in L1210 cells dereased after treating with dexamethasone, but did not be changed with the different concentrations and different times of dexmethason.
The transcription level of Notch gene in CLL patients significantly higher than that in normal controls. The Notch1 protein expression is down-regulated in process of inhibiting L1210 cell proliferation by Ara-C and dexmethason. Notch signaling pathway may mediated anti-apoptosis and drug resistance of CLL cells. Notch molecule possibly plays an important role in the anti-apoptosis and drug-resistance of CLL cells.
研究Notch基因在慢性淋巴细胞白血病细胞中的表达情况，探讨阿糖胞苷或地塞米松治疗后Notch蛋白的变化，以及Notch介导慢性淋巴细胞白血病细胞抗凋亡和耐药的机制。
收集24例慢性淋巴细胞白血病患者及14例健康供者骨髓或外周血中的单个核细胞，采用实时荧光定量PCR（qRT-PCR）检测转录水平上Notch基因、BCL-2以及NF-κB基因的表达。通过蛋白质印迹法检测阿糖胞苷和地塞米松治疗后L1210细胞系中Notch蛋白的变化。
慢性淋巴细胞白血病组中Notch1、Notch2、BCL-2和NF-κB基因的mRNA表达水平显著高于健康对照组（分别为0.8556±0.8726对0.6731±0.5334，P = 0.0182；1.2273±0.8207对0.6577±0.6424，P<0.0001；8.0960±7.5661对0.5969±0.4976，P<0.0001；1.0966±0.6925对0.5373±0.7180, P<0.0001），但Notch3和Notch4基因之间无显著差异（分别为1.1914±2.4219对0.8713±0.7937，P = 0.3427；0.8174±1.0869对0.9752±1.3446，P = 0.2402）。低、中浓度阿糖胞苷处理后L1210细胞中Notch1蛋白表达显著降低，但高浓度阿糖胞苷处理或中浓度阿糖胞苷延长处理时间后表达增加。地塞米松处理后L1210细胞中Notch1蛋白表达降低，但不同浓度和不同时间的地塞米松处理后无变化。
慢性淋巴细胞白血病患者Notch基因转录水平显著高于正常对照。阿糖胞苷和地塞米松抑制L1210细胞增殖过程中Notch1蛋白表达下调。Notch信号通路可能介导慢性淋巴细胞白血病细胞抗凋亡和耐药。Notch分子可能在慢性淋巴细胞白血病细胞抗凋亡和耐药中起重要作用。
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