南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (03): 406-.

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脑胶质瘤细胞糖酵解表型特征及对细胞增殖与凋亡的影响

徐高峰,白晓斌,王茂德,谢万福,李瑞春,李传坤   

  • 出版日期:2013-03-20 发布日期:2013-03-20

Characterization of glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and its association with cell proliferation and apoptosis

  • Online:2013-03-20 Published:2013-03-20

摘要: 目的探讨胶质瘤细胞株SHG44糖酵解表型特征及对细胞增殖、凋亡的影响。方法通过化学模拟法(CoCl2)建立人脑胶
质瘤细胞株SHG44体外缺氧模型。Realtime-PCR、Western blot分别检测HIF-1α mRNA、PDK1 mRNA、PKM2 mRNA、LDHA
mRNA和蛋白的表达;生物发光法检测细胞内ATP值;酶法检测上清中乳酸含量;MTT法检测细胞增殖、流式细胞技术检测细
胞凋亡。结果缺氧条件下,人脑胶质瘤SHG44细胞HIF-1α和糖酵解酶的mRNA和蛋白表达显著升高,细胞内ATP水平下降、
细胞增殖能力减弱、细胞凋亡增加。结论缺氧能诱使肿瘤细胞出现糖酵解表型特征。肿瘤细胞增殖、凋亡与其代谢特征有关。

Abstract: Objective To investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the
association between cell proliferation and apoptosis and the metabolic status. Methods An in vitro hypoxic cell model was
established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of
hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular
ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and
apoptosis of the cells were examined using MTT assay and flow cytometry, respectively. Results The cells in hypoxic culture
showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as
compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an
increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential.
Conclusion Hypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell
death is directly correlated with their metabolic status.