stigmasterol and Liver-Neoplasms

Saringosterol acetate (SSA) was isolated from an edible brown alga Hizikia fusiforme. In this study, we developed an adult zebrafish human hepatocellular carcinoma (HCC) xenograft model to confirm that SSA inhibits tumor growth and metastasis. Established Hep3B cells labeled with the fluorescent tracker CM-Dil were xenografted into the abdominal cavity of zebrafish once every three days for one month. Compared with the control group, the fish injected with Hep3B showed a significant increase in α-fetoprotein (AFP) and factors related to tumor growth and metastasis (IL-6, TNF-α, TGFβ, MMP2, and MMP9). Using the model, it was proven that SSA affected survival rate, AFP production, and the levels of factors related to tumor growth and metastasis via the PI3K/AKT/mTOR and TGFβ/Smad pathways. In conclusion, this HCC model can be used for in vivo experiments to investigate the inhibition of cancer, and SSA may be useful for the treatment of cancer.

Topics: Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Cell Line, Tumor; Female; Humans; Liver Neoplasms; Male; Neoplasm Metastasis; Sargassum; Signal Transduction; Stigmasterol; Xenograft Model Antitumor Assays; Zebrafish

Despite several phytochemical studies of Premna serratifolia Linn. (Verbenaceae), the isolation of active constituents of this plant remains to be explored.. The study isolates cytotoxic terpenoids and steroids from the leaves of Premna serratifolia.. Unsaponifiable matter of hexane soluble fraction obtained from methanol extract was subjected to isolation by column chromatography and preparative TLC. Three compounds PS-01 A, PS-01B and PS-02 A were isolated. PS-01 A and PS-01B were identified by comparative TLC with authentic marker compounds followed by NMR analysis. Further PS-01B was analyzed by HR-GCMS. PS-02 A was subjected to HR-LCMS. All isolated compounds/fractions were evaluated for cytotoxic activity by BSL bioassay and using cell lines A549, HepG2 and L6.. Bioactivity guided fractionation of Premna serratifolia leaves succeeded into isolation of two terpenoids and one steroid compound with significant cytotoxic activity. Here we report the isolation of these cytotoxic terpenoids/steroids from this plant for the first time which could be developed as anticancer agents.

Topics: A549 Cells; Animals; Antineoplastic Agents, Phytogenic; Cell Survival; Chromatography, Thin Layer; Dose-Response Relationship, Drug; Gas Chromatography-Mass Spectrometry; Hep G2 Cells; Hexanes; Humans; Inhibitory Concentration 50; Liver Neoplasms; Lung Neoplasms; Magnetic Resonance Spectroscopy; Methanol; Muscle Fibers, Skeletal; Oleanolic Acid; Phytotherapy; Plant Extracts; Plant Leaves; Plants, Medicinal; Rats; Solvents; Stigmasterol; Verbenaceae

Quercetin (QT) is a potential chemotherapeutic drug with low solubility that seriously limits its clinical use. The aim of this study was enhancing cellular penetration of QT by sterol containing solid lipid nanoparticles (SLNs) which make bilayers fluent for targeting hepatocellular carcinoma cells. Three variables including sterol type (cholesterol, stigmasterol and stigmastanol), drug and sterol content were studied in a surface response D-optimal design for preparation of QT-SLNs by emulsification solvent evaporation method. The studied responses included particle size, zeta potential, drug loading capacity and 24 h release efficiency (RE24%). Scanning electron and atomic force microscopy were used to study the morphology of QT-SLNs and their thermal behavior was studied by DSC analysis. Cytotoxicity of QT-SLNs was determined by MTT assay on HepG-2 cells and cellular uptake by fluorescence microscopy method. Optimized QT-SLNs obtained from cholesterol and QT with the ratio of 2:1 that showed particle size of 78.0 ± 7.0 nm, zeta potential of -22.7 ± 1.3 mV, drug loading efficiency of 99.9 ± 0.5% and RE24 of 56.3 ± 3.4%. IC50 of QT in cholesterol SLNs was about six and two times less than free QT and phytosterol SLNs, respectively, and caused more accumulation of QT in HepG2 cells. Blank phytosterol SLNs were toxic on cells.

Topics: Antineoplastic Agents, Phytogenic; Carcinoma, Hepatocellular; Chemistry, Pharmaceutical; Cholesterol; Hep G2 Cells; Humans; Liposomes; Liver Neoplasms; Nanoparticles; Particle Size; Quercetin; Sitosterols; Stigmasterol

Several studies in humans have demonstrated the hypocholesterolemic effect of plant sterol consumption. It is unclear whether plant sterols regulate lipoprotein metabolism in the liver and intestines, thereby decreasing the levels of circulating atherogenic lipoproteins. We investigated the effect of the three main phytosterols: stigmasterol, campesterol, and beta-sitosterol on lipoprotein production in HepG2 human liver cells and Caco2 human intestinal cells and the mechanisms involved. Cells were incubated for 24h with 50 micromol/L of the different phytosterols or 10 micromol/L of atorvastatin. Very low-density lipoprotein levels (measured by apolipoprotein (apo) B100) in HepG2 cells and chylomicron levels (measured by apoB48) in Caco2 cells were measured using western blotting. Intracellular cholesterol levels were measured using gas chromatography. Analysis was carried out using Student's t-test and ANOVA. Secretion levels of apoB100 significantly decreased by approximately 30% after incubation with all phytosterols compared to control. In addition, cholesterol ester (CE) concentrations significantly decreased when HepG2 cells were incubated with the phytosterols compared to control cells. Secretion of apoB48 from intestinal cells significantly decreased by 15% with stigmasterol, 16% with campesterol and 19% beta-sitosterol compared to control. Collectively the data suggests that plant sterols limit lipid (CE) availability in cells. Decreases in circulating levels of LDL and chylomicron remnants seen in humans with the consumption of margarine phytosterols are possibly due to their effect on lipid production in cells and would therefore reduce the risk of developing cardiovascular disease.

Topics: Anticholesteremic Agents; Apolipoprotein B-100; Apolipoprotein B-48; Apolipoproteins B; Atherosclerosis; Atorvastatin; Caco-2 Cells; Carcinoma, Hepatocellular; Cholesterol; Drug Synergism; Enterocytes; Hepatocytes; Heptanoic Acids; Humans; Liver Neoplasms; Margarine; Phytosterols; Pyrroles; Sitosterols; Stigmasterol