ABSTRACT
Plant sterols have been widely used as chemotherapeutic agents for colorectal cancer for years together. In this study, a novel phytosterol was isolated and characterized from the leaf extract of a medicinal plant, Datura inoxia and was coined as RinoxiaB (RB). This phytosterol was observed to have antiproliferative activity against human colon adenocarcinoma cells, HCT 15. The cell viability assay revealed the IC50 value of the RB as 4 µM. Moreover, RB treated cells showed prominent morphological changes dose dependently and progressively increased the number of dead cells. Additionally, results of the comet, flow cytometry, and cell cycle analysis revealed that the majority of cells were arrested in their S and G2/M phase by blocking the mitotic spindle formation. The western blot analysis (Bcl-2, BAX, Cytochrome C, Caspases 9 & 3) clearly indicated that RB has the ability to induce apoptosis by significantly upregulating (P < 0.05) Bcl-2 and causing mitochondrial damage leading to Cytochrome C release and activation of caspases, which subsequently results in downregulation of BAX expression in the cytosol. Furthermore, the expression of tumor suppressors (p53 and p21) and cell cycle regulatory proteins (Cyclins D1 & B1) suggested that RB inhibit cell proliferation. Thus, the present finding concludes that RB can offer possible apoptotic effects by targeting BAX/Bcl2 pathway in HCT 15 cells, thus alleviating colon cancer.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Colonic Neoplasms/drug therapy , Datura/chemistry , Phytosterols/pharmacology , Plant Extracts/pharmacology , Proto-Oncogene Proteins c-bcl-2/antagonists & inhibitors , bcl-2-Associated X Protein/antagonists & inhibitors , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Molecular Structure , Phytosterols/chemistry , Phytosterols/isolation & purification , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Proto-Oncogene Proteins c-bcl-2/metabolism , Structure-Activity Relationship , bcl-2-Associated X Protein/metabolismABSTRACT
Gold nanoparticles are considered of great importance compared to other noble metal nanoparticles and its wide range of applications like pharmaceutics, therapeutics and diagnostics etc. During the past decade, phytosynthesized gold nanoparticles (AuNPs) are more focused in in vitro and in vivo study. The present study was focused on the gold chloride and phytosynthesized gold nanoparticles from aqueous leaf extract of Cassia roxburghii and their toxic effects on African green monkey normal kidney Vero cell line and three different cancer cell lines such as HepG2, MCF7 and HeLa. Phytosynthesized AuNPs were characterized by HRTEM, EDX, XRD and FTIR analysis. The particles size range of 25-35nm was confirmed by HRTEM. The elemental gold and the crystalline nature of AuNPs were confirmed by EDX and XRD, respectively. The reduction of functional groups was confirmed by FTIR. In in vitro study, the IC50 of HepG2 cells was found to be 30µg/ml compared to other cell lines, HeLa and MCF7 cell line showing IC50 of 50µg/ml and normal Vero cell line also nontoxic up to 75µg/ml confirmed by MTT assay. Further, apoptosis in HepG2 was analyzed by fluorescence microscope and DNA fragmentation was observed in HepG2 treated cells. These results suggested that phytosynthesized AuNPs of C. roxburghii extract clearly limited toxic on normal cells but toxic in cancer cells.
Subject(s)
Cassia/metabolism , Gold/chemistry , Metal Nanoparticles/chemistry , Plant Leaves/metabolism , Animals , Cell Line , Cell Line, Tumor , Chlorocebus aethiops , DNA Fragmentation , Humans , Microscopy, Fluorescence , Spectrum Analysis , Vero CellsABSTRACT
Metallic nanoparticles are major concern, particularly silver nanoparticles (AgNPs) are used in various applications. In the present investigation, we report a novel strategy with biological approach for synthesis of AgNPs using sodium para-hydroxybenzoate tetrahydrate (SPHT) isolated from Vitex negundo leaves. The synthesized SPHT-AgNPs were characterized by UV-vis spectroscopy, high resolution transmission electron microscopy (HRTEM) with selected area electron diffraction (SAED) pattern, field emission scanning electron microscopy (FESEM) with energy-dispersive X-ray spectroscopy (EDX), zeta potential and Fourier transform infrared spectroscopy (FT-IR) analysis. The various pH and temperature were evaluated to find their stability effects on SPHT-AgNPs synthesis peak at 430 nm. The size of SPHT-AgNPs were ranging from 26 to 39 nm and were spherical in shape. The hydroxyl and carboxylic functional groups from bio-reducing mediators of SPHT have a stronger ability towards synthesis of AgNPs, which was confirmed using FT-IR spectrum. In addition, anticancer activity were determined by MTT assay, Annexin V-FITC/PI and cell cycle analysis.
