CD147/HAb18G单克隆抗体对人脑胶质瘤细胞乳酸转运及细胞生长的影响

CD147/HAb18G monoclonal antibody suppresses glycolysis lactic acid metabolism in human glioma cells in vitro

目的研究CD147/HAb18G基因工程单克隆抗体对体外培养U251细胞单羧酸转运蛋白-1(monoc arboxylatetransporter-1,MCT1)表达分布和糖酵解乳酸代谢的影响。方法体外培养U251细胞分为对照组、低剂量组和高剂量组,分别加入不同剂量的CD147/HAb18G单抗封闭细胞表面CD147分子。免疫荧光法检测各组细胞膜上MCT1表达分布改变,分光光度计检测各组细胞内乳酸含量,分别绘制生长曲线并记录生长数据。结果低剂量组和高剂量组MCT表达分布改变,相比对照组其胞膜有效表达减少[随机计数500个细胞中胞膜完整表达细胞数:对照组(367.34±6.92),低剂量组(161.92±7.33),高剂量组(54.58±4.80),P<0.01],胞质无效表达增多[胞质表达细胞数:对照组(12.08±3.03),低剂量组(70.83±4.71),高剂量组(264.58±6.14),P<0.01];细胞内乳酸浓度在低、高剂量组明显增高[对照组(0.222±0.020)mmol/L,低剂量组(0.418±0.015)mmol/L,高剂量组(0.713±0.018)mmol/L,P<0.01],生长指标抑制,生长曲线低平[对照组最大细胞密度(5.24±0.37)×105/ml,低剂量组(4.23±0.48)×105/ml,高剂量组(3.19±0.46)×105/ml,P<0.01]。以上改变在低剂量组和高剂量组间差异亦有统计学意义(P<0.05)。结论 CD147/HAb18G单抗通过干扰CD147对MCT1表达的引导使胶质瘤细胞膜上MCT1有效分布减少,影响其糖酵解产生乳酸的排出,具有抑制肿瘤生长作用。

Objective To explore the effects of CD147/HAb18G genetic engineering monoclonal antibody on the distribution of monocarboxylate transporter-1（MCT1）in U251 cells,and its influence on lactic metabolism of glycolysis.MethodsU251 glioblastoma cells were divided into control（10 mg/L human IgG 0.5 ml）,low dose（5 mg/L CD147/HAb18G Mab 0.5 ml）and high dose（10 mg/L CD147/HAb18G Mab 0.5 ml）groups.The change of MCT1 expression and distribution in each group were demonstrated by immunofluorescence staining.Intracellular lactic acid content were detected by spectrophotometer,and cell growth curves and data of each group recorded for statistical comparison.ResultsThe expression and distribution of MCT1 were significantly changed in low and high dose groups.Compared with control group,valid expression of MCT1 on cell membrane was decreased（random observation of 500 cells for each sample,the cell counts of completed membrane expression in contrast group：367.34±6.92,low dose group：161.92±7.33,high dose group：54.58±4.80,P0.01）,while invalid accumulation of MCT1 in plasma was increased（the cell counts of plasma expression in control group 12.08±3.03,low dose group 70.83±4.71,high dose group 264.58±6.14,P0.01）.Intracellular lactic acid content was significantly enhanced（the lactic acid content in contrast group：0.222±0.020 mmol/L,low dose group：0.418±0.015 mmol/L,high dose group：0.713±0.018 mmol/L,P〈0.01）.Growth curves and data also showed difference among 3 groups [the mean max cell density in control group：（5.24±0.37）×10^5/ml,low dose group：（4.23±0.48）×10^5/ml,high dose group：（3.19±0.46）×105/ml,P〈0.01].The variations mentioned above also had statistical difference between low and high dose group（P〈0.05）.Conclusion CD147/HAb18G monoclonal antibody reduces valid expression of MCT1 on cell membrane by interfering CD147 guiding interactions with MCT1,and restrains the drainage of intracellular lactic acid generated by glycolysis.It has suppression on tumor cell growth.