Heregulin-β1诱导的糖酵解对乳腺癌细胞MCF7迁移的影响

Heregulin-β1-induced Glycolysis Promotes Migration of Breast Cancer Cell Line MCF7

目的探讨heregulin-β1(HRG-β1)对糖酵解的诱导作用及其诱导的糖酵解在乳腺癌细胞MCF7迁移中的作用。方法用PBS(对照组)或HRG-β1处理MCF7细胞12、24和48h,或HRG-β1+草氨酸盐(oxamate,OX)联用处理MCF7细胞24h,收集培养基测定葡萄糖消耗量和乳酸生成量,收集细胞用Western blot检测乳酸脱氢酶A(lactate dehydrogenase A,LDHA)蛋白的表达。用PBS(对照组)、HRG-β1或HRG-β1+OX联用处理MCF7细胞48h,划痕实验检测伤口愈合率以反映细胞的迁移能力。结果HRG-β1处理MCF7细胞12、24和48h组的葡萄糖消耗量、乳酸生成量和LDHA蛋白水平增加均在24h达最大值,与对照组比较差异有统计学意义(P<0.05);与HRG-β1诱导组比较,HRG-β1+OX联用组的葡萄糖消耗量差异无统计学意义(P>0.05),乳酸生成量降低(P<0.01),LDHA蛋白表达量减少(P<0.05);MCF7细胞划痕48h后,对照组和HRG-β1+OX联用组的伤口愈合率相当(P>0.05),均低于HRG-β1诱导组,差异有统计学意义(P<0.001)。结论 HRG-β1通过上调LDHA诱导糖酵解从而促进乳腺癌细胞MCF7的迁移。

Objective To explore whether heregulin-β1（HRG-β1）can induce glycolysis and the role of HRG-β1-induced glycolysis in the migration of human breast cancer cell line MCF7.Methods MCF7 cells were treated with PBS（PBS group）or HRG-β1for 12,24 and 48h.Culture media were harvested for glucose uptake and lactate production assays,and cells were collected and lactate dehydrogenase A（LDHA）protein levels were detected by using Western blot.MCF7 cells were treated with PBS（PBS group）,HRG-β1or HRG-β1plus oxamate（OX）for 24 h.Culture media were harvested for glucose uptake and lactate production assays,and cells were harvested and the protein levels of LDHA was detected by Western blot.The wound healing assay was used to detect the migration of MCF7 cells treated with PBS（PBS group）,HRG-β1or HRG-β1plus OX for 48 h.ResultsMCF7cells treated with HRG-β1for 12,24 and 48hdisplayed higher levels of glucose uptake,lactate production and LDHA protein levels when the levels reached the peak at 24 h.The differences of glucose uptake,lactate production and LDHA protein levels between PBS group and HRG-β1group were statistically significant（P〈0.05）.Compared to HRG-β1group,the glucose uptake of HRG-β1plus OX treated group was not significantly different（P〉0.05）,but the statistically significant decrease of lactate production and LDHA protein levels were noticed（P〈0.01 and P〈0.05）.When MCF7 cells were scratched for 48 h,the wound healing rate of control group,HRG-β1group and HRG-β1plus OX group was（49±5.09）%,（100±2.21）% and（51±4.10）%respectively.The difference of each group was statistically significant（P〈0.001）.Conclusion HRG-β1induces glycolysis via upregualtion of LDHA and HRG-β1-induced glycolysis promotes the migration of breast cancer cells line MCF7.