[Heregulin-β1诱导的糖酵解促进乳腺癌细胞系MCF7的迁移]

Jiang Su-Fang, Tu Kai-Ling, Zhou Lan, Fu Qiang, Zhao Yu-Hua

Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 60041, China.

Department of Anatomy, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 60041, China.

Sichuan Da Xue Xue Bao Yi Xue Ban. 2016 Jul;47(4):463-467.

OBJECTIVES

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-β1 for 12, 24 and 48 h. 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-β1 or HRG-β1 plus 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-β1 or HRG-β1 plus OX for 48 h.

RESULTS

MCF7 cells treated with HRG-β1 for 12, 24 and 48 h displayed 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-β1 group were statistically significant ( <0.05). Compared to HRG-β1 group, the glucose uptake of HRG-β1 plus OX treated group was not significantly different ( >0.05), but the statistically significant decrease of lactate production and LDHA protein levels were noticed ( <0.01 and <0.05). When MCF7 cells were scratched for 48 h, the wound healing rate of control group, HRG-β1 group and HRG-β1 plus OX group was (49±5.09)%, (100±2.21)% and (51±4.10)% respectively. The difference of each group was statistically significant ( <0.001).

CONCLUSIONS

HRG-β1 induces glycolysis via upregualtion of LDHA and HRG-β1-induced glycolysis promotes the migration of breast cancer cells line MCF7.

目的

探讨神经调节蛋白-β1（HRG-β1）是否能诱导糖酵解以及HRG-β1诱导的糖酵解在人乳腺癌细胞系MCF7迁移中的作用。

方法

将MCF7细胞用PBS（PBS组）或HRG-β1处理12、24和48小时。收集培养基进行葡萄糖摄取和乳酸生成测定，并收集细胞，用蛋白质免疫印迹法检测乳酸脱氢酶A（LDHA）蛋白水平。将MCF7细胞用PBS（PBS组）、HRG-β1或HRG-β1加草氨酸盐（OX）处理24小时。收集培养基进行葡萄糖摄取和乳酸生成测定，并收集细胞，用蛋白质免疫印迹法检测LDHA的蛋白水平。采用划痕实验检测用PBS（PBS组）、HRG-β1或HRG-β1加OX处理48小时的MCF7细胞的迁移情况。

结果

用HRG-β1处理12、24和48小时的MCF7细胞表现出较高的葡萄糖摄取、乳酸生成水平和LDHA蛋白水平，且在24小时时达到峰值。PBS组和HRG-β1组之间的葡萄糖摄取、乳酸生成和LDHA蛋白水平差异具有统计学意义（<0.05）。与HRG-β1组相比，HRG-β1加OX处理组的葡萄糖摄取无显著差异（>0.05），但乳酸生成和LDHA蛋白水平有统计学意义的下降（<0.01和<0.05）。当MCF7细胞划痕48小时时，对照组、HRG-β1组和HRG-β1加OX组的伤口愈合率分别为（49±5.09）%、（100±2.21）%和（51±4.10）%。各组间差异具有统计学意义（<0.001）。