U251细胞MCT1、4在氧糖剥夺后的表达变化及其意义

发布时间：2019-01-20 15:27

【摘要】：目的:观察氧糖剥夺(oxygen-glucose deprivation,OGD)后,神经胶质瘤细胞U251的乳酸转运和单羧酸转运蛋白(monocarboxylate transporter,MCT)1、4的表达变化,以探讨胶质瘤增殖生长的分子机制。方法:U251细胞随机分为对照组和模型组(OGD组),模型组的干预时间点为1、3、6 h。四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium,MTT)检测细胞增殖生长情况,HE染色观察细胞的形态变化,乳酸试剂盒检测细胞培养液中的乳酸含量,免疫荧光和免疫印迹观察MCT1和MCT4的表达变化情况。结果:与对照组相比,随着氧糖剥夺时间延长,模型组细胞增殖能力下降(对照组、OGD 1 h、OGD 3 h、OGD 6 h组的光密度值分别为0.584±0.031、0.537±0.046、0.441±0.079和0.200±0.011),细胞培养液中的乳酸浓度增高(对照组、OGD 1 h、OGD 3 h、OGD 6 h组乳酸浓度值分别为0.150±0.029、0.707±0.022、1.167±0.024和1.325±0.023)。MCT1、4的表达上调,并随氧糖剥夺时间延长其表达逐渐增强(MCT1:F=46.256,P=0.000;MCT4:F=247.400,P=0.000)。细胞培养液中加入MCTs的抑制剂4-CIN后,其乳酸浓度降低(对照组、150μmol/L组、425μmol/L组的乳酸浓度值分别为0.249±0.012、0.222±0.016、0.167±0.117)。结论:U251细胞氧糖剥夺后,随着细胞培养液中乳酸浓度增加,MCT1、4亦表达上调,而细胞的增殖生长能力下降;抑制MCT的表达,细胞培养液中的乳酸浓度降低,提示MCT1、4可能通过参与神经胶质瘤缺血缺氧微环境中乳酸的转运,进而调控细胞的增殖生长过程。
[Abstract]:Aim: to investigate the changes of lactate transport and monocarboxylate transporter (monocarboxylate transporter,MCT) 1 / 4 expression in glioma cell line U251 after oxygen glucose deprivation (oxygen-glucose deprivation,OGD) in order to explore the molecular mechanism of glioma proliferation and growth. Methods: U251 cells were randomly divided into control group and model group (OGD group). (methyl thiazolyl tetrazolium,MTT) was used to detect cell proliferation and growth, HE staining was used to observe the morphological changes of cells, and lactic acid kit was used to detect the content of lactic acid in cell culture medium. Immunofluorescence and Western blot were used to observe the expression of MCT1 and MCT4. Results: compared with the control group, the proliferation ability of the model group decreased with the prolongation of oxygen glucose deprivation time (control group, OGD 1 h, OGD 3 h). The optical density of OGD for 6 h was 0.584 卤0.031 卤0.046 卤0.441 卤0.079 and 0.200 卤0.011, respectively. The concentration of lactic acid in cell culture medium was increased (control group, OGD 1 h, OGD 3 h). The concentration of lactic acid in OGD 6 h group was 0.150 卤0.029 卤0.707 卤0.022 卤1.167 卤0.024 and 1.325 卤0.023, respectively. The expression of MCT1,4 was up-regulated and gradually increased with the prolongation of oxygen glucose deprivation time (MCT1:F=46.256,P=0.000;). MCT4:F=247.400,P=0.000) The concentration of lactic acid decreased after the addition of 4-CIN, an inhibitor of MCTs (control group, 150 渭 mol/L group, 425 渭 mol/L group), the lactate concentration was 0.249 卤0.012 卤0.222 卤0.016 渭 mol/L 0.167 卤0.117, respectively. Conclusion: after U251 cells were deprived of oxygen sugar, the expression of MCT1,4 was up-regulated with the increase of lactic acid concentration in the culture medium, but the proliferation and growth ability of U251 cells decreased. The inhibition of MCT expression and the decrease of lactic acid concentration in cell culture medium suggest that MCT1,4 may regulate the proliferation and growth of glioma cells by participating in the transport of lactic acid in hypoxia-ischemia microenvironment.
【作者单位】：重庆医科大学神经科学中心重庆医科大学基础医学院解剖教研室;
【基金】：国家自然科学基金资助项目(编号:81671312、81000566) 重庆市基础与前沿研究计划资助项目(编号:cstc2015jcyj A10018)
【分类号】：R739.41

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