Jar-TTA, a natural diterpenoid derivatives, induces apoptosis in human esophageal cancer cells through dual inhibition of glycolysis and oxidative phosphorylation / 中国药理学通报

ABSTRACT

Aim To investigate the effects of Jar-TTA in dual inhibition of glycolysis/oxidative phosphoryla-tion in human esophageal cancer cells and explore the related molecular mechanism. Methods The effect of Jar-TTA on the esophageal cancer cell EC 109 and KYSE-150 viability was examined using MTT assay. The effect of Jar-TTA in apoptosis morphology and mitochondrial membrane potential ( MMP) was observed by a fluorescence microscopy. The apoptosis of cell lines treated with Jar-TTA, the quantitative analysis of MMP falling, as well as the glucose uptake was ana-lyzed by flow cytometry. The mitochondrial OXPHOS and glycolysis of EC 109 cells in response to Jar-TTA were analyzed using a Seahorse XFp extracellular flux analyzer by real-time measurements of the oxygen consumption rate (OCR, indicative of mitochondrial OXPHOS) and extracellular acidification rate(ECAR, indicative of glycolysis). The expression of the proteins related with glycolysis were detected by Western blot. Results Jar-TTA caused strong antiproliferation in EC 109 and KYSE-150 cells in a concentration-dependent manner. 2 , 4 and 8 jimol • L"1 Jar-TTA treat-ments of EC 109 cells for 24 h resulted in a significant increase of early apoptosis population up to (27. 9 ± 6. 1)%, (71.1 ±9.3)% and (65. 0 ±9.5)%, respectively , compared to control treated cells (5. 6 ± 3.2)%. The mitochondrial OXPHOS and glycolysis were significantly inhibited in EC 109 incubated by 4 and 8 p,mol • L"1 Jar-TTA for 2 h. In addition, Jar-TTA induced the drop of MiMP. Furthermore, the glucose uptake and the expression of GLUT4 and LDHA were distinctly inhibited in EC 109 treated by Jar-TTA. Conclusions Jar-TTA induces apoptosis of human e-sophageal cancer cells through dual inhibition of glycolysis and oxidative phosphorylation, which is related with the drop of MMP collapse, the decrease of glucose uptake and the down-regulation of GLUT4 and LDHA in EC 109 treated by Jar-TTA.