Simultaneous use of erythropoietin and LFM‐A13 as a new therapeutic approach for colorectal cancer

Abstract

Background and PurposeBruton's tyrosine kinase (Btk) is a non‐receptor tyrosine kinase involved in the activation of signalling pathways responsible for cell maturation and viability. Btk has previously been reported to be overexpressed in colon cancers. This kind of cancer is often accompanied by anaemia, which is treated with an erythropoietin supplement. The goal of the present study was to assess the effects of combination therapy with erythropoietin β (Epo) and LFM‐A13 (Btk inhibitor) on colon cancer in in vitro and in vivo models.Experimental ApproachDLD‐1 and HT‐29 human colon adenocarcinoma cells were cultured with Epo and LFM‐A13. Cell number and viability, and mRNA and protein levels of Epo receptors, Btk and Akt were assessed. Nude mice were inoculated with adenocarcinoma cells and treated with Epo and LFM‐A13.Key ResultsThe combination of Epo and LFM‐A13 mostly exerted a synergistic inhibitory effect on colon cancer cell growth. The therapeutic scheme used effectively killed the cancer cells and attenuated the Btk signalling pathways. Epo + LFM‐A13 also prevented the normal process of microtubule assembly during mitosis by down‐regulating the expression of Polo‐like kinase 1. The combination of Epo and LFM‐A13 significantly reduced the growth rate of tumour cells, while it showed high safety profile, inducing no nephrotoxicity, hepatotoxicity or changes in the haematological parameters.Conclusion and ImplicationsEpo significantly enhances the antitumour activity of LFM‐A13, indicating that a combination of Epo and LFM‐A13 has potential as an effective therapeutic approach for patients with colorectal cancer.

The intensification of anticancer activity of LFM-A13 by erythropoietin as a possible option for inhibition of breast cancer

“…In this study, we not only confirmed the antiproliferative action of LFM-13 in both breast cancer cells lines but also indicated that the addition of Epo to this system further reduced viability of these cells. These results are consistent with our previous findings, which demonstrated that Epo appears to be a chemosensitizer and showed a synergistic antiproliferative effect with LFM-A13 in two colon cancer cells DLD-1 and HT-29 23 . The high effectiveness of the combination used has also been confirmed in zebrafish embryo xenograft model.…”

“…The fish were incubated with Epo þ LFM-A13 at the highest doses for 48 h. The control group received the solvent LFM-A13 (10% DMSO/PBS). In both MCF-7 and MDA-MB- 23 xenografts we observed significant reduction in tumour development in Epo þ LFM-A13 treated group compared with the control (Figure 1(F)). As shown in Figure 1(G 13 .…”

“…We observed a significant antiproliferative effect exerted by the combination of Epo þ LFM-A13 in this model. BTK is responsible for the proliferation and survival of cancer cells 13 , it plays a key role in solid tumour development, including ovarian 25 , prostate 8 , and colon cancer 23 . It was also shown that BTK is required for efficient signal transduction by the ligand-activated EpoR.…”

Erythropoietin Intensifies the Proapoptotic Activity of LFM-A13 in Cells and in a Mouse Model of Colorectal Cancer

“…Animal studies are reported in compliance with the ARRIVE guidelines [ 47 ]. Xenograft animal models were obtained as described previously [ 14 ] using DLD-1 and HT-29 cells (50 μL suspension containing 1 × 10 8 cells for each animal, Figure 5 a). In previous studies, we showed no effect of the Epo solvent on the number of colon cancer cells [ 20 ] and, in accordance with the 3R, we reduced the number of animals [ 48 ].…”

“…It is well known that Epo used to treat anemia promotes survival, proliferation, and differentiation of the progenitors of erythropoiesis, exerting a proangiogenic and antiapoptotic effect [ 12 , 13 ]. Despite this, the results of our recent study indicate that Epo intensifies the antiproliferative activity of LFM-A13 [ 14 ].…”