Expression of EGFR/HER2 proteins and activation of connected transducers in BTC cell lines Expression of EGFR and HER2 proteins at the same time as their molecular pathways have been evaluated by western blot ana lysis on four BTC cell lines. As shown in figure 3, all cell lines expressed EGFR and HER2 receptors. Particularly, EGI 1 cell line expressed substantial ranges of EGFR and HER2 proteins and minimal levels of PTEN. TGBC1 TKB cell line expressed higher degree of phosphorylated Akt, mTOR and MAPK suggesting sustained activation of those pathways. Moreover, HER2 membrane expression was evaluated by immunocitochemistry. ICC cell line HuH28 showed the highest HER2 membrane expression, scored 3, ECC cell lines EGI 1 and TFK 1 were scored 1, whereas GBC cell line TGBC1 TKB showed the lowest HER2 expres sion. After 72 h of treatment, everolimus was capable of inhibit mTOR phosphorylation in all BTC cell lines, but did not influence Akt and MAPK phosphorylation.
Sorafenib down regulated MAPK phosphorylation in all cell lines and did not influence mTOR and Akt phos phorylation. Lapatinib slightly down regulated Akt phosphorylation selleck Obatoclax in all BTC cell lines, but not MAPK nor mTOR. Gefitinib down regulated Akt phosphorylation only in EGI 1 cell line whilst erloti nib had no evident effects on Akt/mTOR and MAPK phosphorylation. Antiproliferative impact of gemcitabine and EGFR/HER2 pathway inhibitors in BTC cell lines The antiproliferative effect of various molecular targeted medication blocking EGFR/HER2 receptor or pathways exposed a broad range of response in BTC cell lines. The ICC cell line HuH28 was resis tant to all medication except lapatinib. Lapati nib also inhibited proliferation of EGI 1 and TFK1, while TGBC1 TKB cell line was resistant. selleckchem Everolimus EGFR TKIs had a significant impact on ECC cell lines, but no effect was revealed for the GBC cell line TGBC1 TKB.
The multi kinase inhibitor sorafenib had a substantial efficacy on EGI 1 plus a slight result on TFK1 and TGBC1 TKB. A reduction of 50% of cell growth was obtained that has a comparatively very low median doses of m TOR inhibitor everolimus on TFK1, on EGI 1 and on TGBC1 TKB. The chemotherapeutic agent gemcitabine was hugely efficient on EGI 1, moderate effective on TFK 1 and TGBC1 TKB and ineffective on HuH28. The blend of targeted medicines with gemcitabine permitted a substantial reduction of median dose. Curiosity ingly, erlotinib conferred sensitivity to gemcitabine in HuH28, resistant towards the exact same drug as single agent and also to all other combinations. In other cell lines, the most beneficial consequence was obtained using the chemotherapeutic agent and everolimus, highly effective on extrahepatic cell lines, and gallbladder cell line. For that other combinations, responsiveness depended on cell lines. Discussion The rising of international incidence, bad prognosis and lack of efficient treatment make the management of BTCs more emphasize the need to have of powerful therapeutic agents.