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目的:研究糖酵解抑制剂2-脱氧-D-葡萄糖(2-deoxy-D-glucose,2-DG)对人类非霍奇金淋巴瘤细胞株Namalwa、SU-DHL-4的作用效应及机制。方法:采用锥虫蓝拒染法检测细胞增殖,D-葡萄糖[HK法]检测试剂盒检测葡萄糖浓度,乳酸测试盒检测糖酵解途径终产物乳酸含量,反转录实时定量PCR(RTQ-PCR)检测糖酵解途径关键酶基因的转录水平,流式细胞术检测细胞周期。以健康人外周血梯度离心法分离所得的淋巴细胞为正常对照组。结果:淋巴瘤细胞株Namalwa和SU-DHL-4中糖酵解相关基因HK1、LDHA、GLUT1、HIF-1α在转录水平明显高于正常对照。2-DG能抑制淋巴瘤细胞株葡萄糖消耗,减少乳酸生成,将细胞周期阻滞于G0/G1期,致使细胞增殖受抑。结论:淋巴瘤细胞株Namalwa、SU-DHL-4存在糖酵解异常,2-DG通过抑制淋巴瘤细胞株糖酵解通路,可干扰细胞能量代谢,阻断细胞周期,使细胞增殖受抑,从而发挥抗肿瘤作用。
AIM: To investigate the effect and mechanism of 2-deoxy-D-glucose (2-DG), a glycolysis inhibitor, on human non-Hodgkin’s lymphoma cell lines Namalwa and SU-DHL-4 . Methods: Cell proliferation was detected by trypan blue exclusion method. Glucose concentration was measured by D-glucose [HK method] detection kit. Lactate test kit was used to detect lactic acid content of the end product of the glycolysis pathway. Reverse transcription-PCR (RTQ-PCR) ) To detect the transcriptional level of the key enzyme gene of glycolytic pathway, and the cell cycle was detected by flow cytometry. Lymphocytes isolated by healthy human peripheral blood gradient centrifugation were normal control group. Results: The levels of glycolytic genes HK1, LDHA, GLUT1 and HIF-1α in lymphoma cell lines Namalwa and SU-DHL-4 were significantly higher than those in normal controls. 2-DG can inhibit the glucose consumption of lymphoma cell lines, reduce the formation of lactic acid, the cell cycle arrest in the G0 / G1 phase, resulting in inhibition of cell proliferation. Conclusion: Lymphoma cell lines Namalwa and SU-DHL-4 have abnormal glycolysis. By inhibiting the glycolytic pathway of lymphoma cell lines, 2-DG can interfere with cell energy metabolism, block cell cycle, inhibit cell proliferation, To play an antitumor effect.