Cisplatin suppresses the growth and proliferation of breast and cervical cancer cell lines by inhibiting integrin β5-mediated glycolysis

Abstract

Cancer cells harbor lower energy consumption after rounds of anticancer drugs, but the underlying mechanism remains unclear. In this study, we investigated metabolic alterations in cancer cells exposed to cisplatin. The present study exhibited cisplatin, known as a chemotherapeutic agent interacting with DNA, also acted as an anti-metabolic agent. We found that glycolysis levels of breast and cervical cancer cells were reduced after cisplatin treatment, resulting in cells growth and proliferation inhibition. We demonstrated that cisplatin suppressed glycolysis-related proteins expression, including glucose transporter 1 (GLUT1), glucose transporter 4 (GLUT4) and lactate dehydrogenase B (LDHB), through down-regulating integrin β5 (ITGB5)/focal adhesion kinase (FAK) signaling pathway. ITGB5 overexpression rescued cisplatin-induced inhibition of cancer cell glycolysis, growth and proliferation. Conclusively, we reveal a novel insight into cisplatin-induced anticancer mechanism, suggesting alternative strategies to the current therapeutic approaches of targeting ITGB5, as well as of a combination of cisplatin with glucose up-regulation chemotherapeutic agents to enhance anticancer effect.

Keywords: Cisplatin; ITGB5; glucose metabolism; integrin.

Figure 1

Cisplatin inhibits glycolysis and lactate metabolism of MDA-MB-231 and siha cells. A, B. The values of IC₅₀ of MDA-MB-231 and siha cells treated with cisplatin were determined by CCK8 kit assay. C. Detection of glucose uptake capacity of MDA-MB-231 and siha cells treated with cisplatin by glucose uptake kit (P < 0.001). D. Detection of lactate production capacity of MDA-MB-231 and siha cells treated with cisplatin by lactate production kit (P < 0.001). E, F. Quantitative real-time PCR analysis of the expression levels of GLUT1, GLUT4, and LDHB in MDA-MB-231 or siha cells with or without the treatment of cisplatin (P < 0.05). G. Immunoblotting analysis of GLUT1, GLUT 4 and LDHB proteins in MDA-MB-231 or siha cells with or without the treatment of cisplatin.

Figure 2

ITGB5 rescues the cisplatin-induced inhibition of cell glycolysis. A, B. Quantitative real-time PCR and immunoblotting analysis of ITGB5 expression in MDA-MB-231 and siha cell with the treatment of cisplatin (P < 0.05). Error bars = 95% confidence intervals (CIs). C, D. Detection of the glucose uptake and lactate production of MDA-MB-231 or siha cell expressing ITGB5 cDNA and their controls with or without the treatment of cisplatin (P < 0.05). E, F. The proliferous capacity of MDA-MB-231 or siha cell expressing ITGB5 cDNA and that of their controls with or without the treatment of cisplatin by CCK8 Kit (P < 0.05). Error bars = 95% CIs. G, H. Detection of colonies formation ability of MDA-MB-231 or siha cell expressing ITGB5 cDNA and their controls with or without the treatment of cisplatin and quantitative analysis of colonies formation (P < 0.05). Error bars = 95% CIs.

Figure 3

ITGB5 overcome the adverse metabolic effect of cisplatin on MDA-MB-231 and siha cell lines via FAK/p-FAK pathway. A, B. Quantitative real-time PCR analysis of GLUT1, GLUT4, and LDHB in MDA-MB-231 or siha cell expressing ITGB5 cDNA or their control cells with or without the treatment of cisplatin (P < 0.05). Error bars = 95% CIs. C. Immunoblotting analysis of GLUT1, GLUT 4, LDHB, FAK and p-FAK in MDA-MB-231 or siha cell expressing ITGB5 cDNA or their control cells with or without the treatment of cisplatin.

Figure 4

Xenograft tumor burden in mice with overexpression of ITGB5 with or without the treatment of cisplatin. A-D. In vivo tumorigenesis examined by animal assay and subcutaneous tumor growth from mice injected with cells expressing ITGB5 cDNA and the corresponding controls with or without the treatment of cisplatin (n = 10 for each group, P > 0.05 in ITGB5 overexpressing group). Error bars = 95% CIs. Figures show tumor and tumor weights of mice at the end of observation. E. Immunohistochemtry staining of xenograft tumor tissues. Tissues was stained with rabbit anti-ITGB5, GLUT1, GLUT4 and LDHB antibody and visualized with goat anti-rabbit secondary antibody (Magnification × 400).