Hepcidin TH1-5 Induces Apoptosis and Activate Caspase-9 in MCF-7 Cells.

Breast cancer is the most commonly diagnosed and leading cause of cancer deaths among women globally. In continuation of our investigation into the cytotoxicity of the antimicrobial peptide, Hepcidin TH1-5 on human breast adenocarcinoma cell line (MCF-7), we further affirm the apoptosis-inducing effect of the cysteine-rich peptide in the present study. Annexin V-fluorescein isothiocyanate and propidium iodide (annexin V-FITC/PI) apoptosis assay was performed after treatment of the cells. In the determination of caspase activity and pathway of apoptosis, luminescent assay was also performed where caspase-3/7, caspase-8 and caspase-9 were evaluated. Results of annexin V-FITC/PI staining showed proportion of early apoptotic cell were 73.67 ± 4.93%, 61.00 ± 5.57% and 44.33 ± 2.52% at 24, 48 and 72 hours respectively, while late apoptotic cell were 6.33 ± 1.53%, 23 ± 3.56% and 34 ± 3.51% within the same time interval. Based on the data from the luminescence test, Hepcidin TH1-5 activated caspases-3/7 and -9 which suggests that the apoptosis induced was due to the peptide treatment. Hepcidin TH1-5 induced apoptosis in MCF-7 via the activation of caspase-9 of the intrinsic pathway. These results support our previous findings of the cytotoxicity of Hepcidin TH1-5 and indicate that the peptide may be a potential agent for breast cancer therapy.

Discriminating Ficus deltoidea var. bornensis from Different Localities by HPTLC and FTIR Fingerprinting.

A nondestructive, efficient, and accurate fingerprinting method using Fourier transform infrared spectroscopy (FTIR) has been developed and optimized for the investigation and demonstration of the variance in chemical characteristics among extracts of Ficus deltoidea Jack var. bornensis from different but closely situated origins. The capacity of attenuated total reflectance (ATR) to differentiate these samples were studied using methanol and water extracts which were preliminarily screened using HPTLC with vitexin and isovitexin being used as markers for authentication. The mobile phase used was ethyl acetate: formic acid (0.1%): methanol at ratio of 5:5:2 (v/v/v) and the profile showed that methanol extracts had higher affinity for the markers. The FTIR spectra indicated that there was no obvious difference in spectroscopic pattern for either extracts when comparing samples from different localities but the absorption intensities of some peaks were different. Multivariate statistical analyses of PCA and HCA showed that both these techniques were capable of identifying the most similar as well as most differing samples and the identification depended on the type of extract. Overall, FTIR fingerprinting has the potential to be a fast and reliable analytical methodology for the discrimination between variants of plant from closely situated locations and hence chemically similar samples.