Exposure of Hepatocellular Carcinoma Cells to Ankaferd Blood Stopper® Alters Cell Death Signaling Networks Confirmed by Oncoproteomic and Genomic Profiling Studies

Abstract

Background: Ankaferd Blood Stopper® is a commercially available herbal extract with potent blood-stopping property and is clinically used to treat immediate dental, dermal, external and internal bleeding. Its possible anti-neoplastic effect or whether it ingenerates drug resistance in cells has not been previously scrutinized. Objective: In the present study HEPG2 hepatocellular carcinoma cell line was exposed to clinically used dose (8 μL/mL) of the blood stopper for 24 h and the behavioral changes were investigated on both proteomic and genomic levels. Method: Cell culture experiments were performed by using Ankaferd® application in HepG2 cell line. Cytotoxic activity experiments with MTT, oncoproteomic studies with 2d gel electrophoresis, genomic studies were performed with RT-PCR. Results: It was seen that the agent did not significantly inhibit cell viability subsequent to 24 h of the treatment meanwhile, it clearly deducted cell viability after 72 h. Although reduction of HEPG2 cell proliferation was not witnessed as a response to 24 h of treatment with Ankaferd®, genomic and oncoproteomic analysis demonstrated diversification. Conclusion: It was established that protein processing networks in endoplasmic reticulum which regulate protein folding, relocation and degradation were effected. Additionally, it was proved that along with the elongation of the exposure period, mitochondrial apoptotic pathway may be activated due to hnRNP F-p53 interaction. Given to the fact that the agent did not cause P-glycoprotein-dependent drug resistance unlike many clinically used chemotherapeutic agents, it can also be considered for combination treatment. Overall, these findings suggest that Ankaferd® offers a novel promising approach against hepatocellular carcinoma which needs further investigation.