Btk is non-receptor tyrosine kinases involved in the activation of signaling pathways responsible for maturation and viability of the cells. It plays an important role in the development of B-cell tumors, activating antiapoptotic pathways. Btk has previously been reported to be overexpressed in prostate cancer which correlated with cancer grades. Colorectal cancer is among the five most frequent causes for cancer-related deaths in Europe. This kind of cancer often accompanied by anemia which is treated with erythropoietin supplement. The aim of this study was to assess the effects of combination therapy with erythropoiethin beta (Epo) and LFM-A13 (Btk inhibitor) on colorectal carcinoma cells both in in vitro and in animal models.
DLD-1 and HT-29 human colon adenocarcinoma cells were cutured in medium with Epo and LFM-A13. Cell proliferation was measured with an automated cell counter. Expression of Btk by Western Blotting and Akt mRNA by RT-PCR, and its protein was assessed and confocal microscopy, respectively. Analysis of apoptosis by flow-cytometry was also carried out. Nude mice were inoculated with adenocarcinoma cells and treated with Epo and LFM-A13.
Herein, we found that simultaneous use of Epo and LFM-A13 exert an additive inhibitory effect on colon cancer cell growth. Featured therapeutic scheme resulted in effective cell killing, accompanied by attenuation of Btk, Akt signaling pathway and increased of apoptosis. This combination is also effective
Results of this study show that that adding Epo significantly enhances the antitumor activity of LFM-A13. The results of our study indicate the potential use of a combination of Epo and LFM- A13 as an effective therapeutic approach for colorectal cancer.
Justyna Magdalena Hermanowicz
National Science Centre, Poland No. DEC-2017/01/X/NZ5/00362
All authors have declared no conflicts of interest.