rosmarinic-acid and Carcinoma--Hepatocellular

Rosmarinic acid (RA) is a natural phenolic compound that acts as a Fyn inhibitor by 53 homology modeling of the human Fyn structure. Therefore, the apoptosis mechanism related to  NF-κB signaling pathway induced by RA in HepG2 was investigated.. The cell growth, apoptosis, and proliferation of HepG2 regulated by various concentrations of RA were studied. The proteins expression of MMP-2, MMP-9, PI3K, AKT, NF-κB, and apoptosis-related proteins Bax, Bcl-2, cleaved caspase-3 were detected.. RA significantly reduced proliferation rates, inhibited migration and invasion, and decreased the expressions of invasion-related factors, such as matrix metalloproteinase (MMP)-2 and MMP-9. TUNEL staining revealed that RA resulted in a dose-dependent increase of HepG2 cell apoptosis. In line with this finding, the expression of apoptosis suppressor protein Bcl-2 was downregulated and that of the pro-apoptotic proteins Bax and cleaved caspase-3 was increased. In addition, we found that the phosphatidylinositol 3-kinase (PI3K)/Akt/nuclear factor kappa B (NF-κB) signaling pathway was involved in RA-mediated inhibition of HepG2 cell metastasis.. Our study identified that  RA as a drug candidate for the treatment of HCC.

Topics: Antioxidants; Apoptosis; Carcinoma, Hepatocellular; Cell Movement; Cell Proliferation; Cinnamates; Depsides; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; NF-kappa B; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Rosmarinic Acid; Signal Transduction; Tumor Cells, Cultured

To investigate the effect of rosmarinic acid (RosA) on hepatocellular carcinoma cell in vivo and in vitro and to explore its possible mechanism of anti-hepatocarcinoma.. The hepatocellular carcinoma cell line SMMC-7721 was treated with different concentrations of RosA (0, 20, 50, 100 μmol/L) to detect cell proliferation, cell cycle, apoptosis and invasion.PI3K pathway-specific activator IGF-1 was used to explore whether the mechanism for RosA action relates to PI3K/AKT signal pathway.Nude mice inoculated with SMMC-7721 cells were treated with different doses of RosA (0, 5, 10 and 20 mg/kg) to detect the tumor formation of cancer cells in vivo.. RosA significantly inhibited the proliferation of SMMC-7721 cells and induced G1 arrest and apoptosis in a dose-dependent manner. RosA might inhibit cell invasion by regulating epithelial-mesenchymal transition. Rescue experiments showed that IGF-1 could reverse the inhibition of PI3K/AKT/mTOR signal pathway by RosA and the effect on proliferation, apoptosis, cell cycle, invasion and EMT by IGF-1 in SMMC-7721 cells;RosA could inhibit tumor formation of SMMC-7721 cells in vivo.. RosA can inhibit the proliferation and invasion of hepatocellular carcinoma cell in vitro and inhibit tumour growth in vivo and the mechanism may relate to inhibiting the activation of PI3K/AKT signal pathway.

Topics: Animals; Apoptosis; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cinnamates; Depsides; Humans; Insulin-Like Growth Factor I; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rosmarinic Acid; TOR Serine-Threonine Kinases

BACKGROUND Hepatic carcinoma is the third leading cause of cancer-related deaths. This study aimed to evaluate the anti-tumor effects of rosmarinic acid (RosA) combined with Adriamycin (ADM) on proliferation and apoptosis of hepatic carcinoma cell lines. MATERIAL AND METHODS Human HepG2 and Bel-7402 cells were treated with RosA and ADM and divided into HepG2 or Bel-7402, 25 μg/ml, 50 μg/m, and 100 μg/ml RosA+0.4 μg/ml ADM groups, respectively. The Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell viability. Immunohistochemistry assay was used to examine B cell lymphoma-2 (Bcl-2) and Bcl-2-associated X (Bax) expression. Cell cycle analysis was used to detect cell cycle distribution. Flow cytometry and terminal deoxynucleotidyl transferase-mediated d-UTP nick-end labeling (TUNEL) assay were utilized to evaluate apoptosis. RESULTS RosA combined with ADM damaged cell morphology and decreased cell viability, and significantly decreased S-phase cell numbers compared to the HepG2 or Bel-7402 group (p<0.05). Apoptosis rates in the RosA combined with ADM group were significantly increased compared to the HepG2 or Bel-7402 group (p<0.05). TUNEL assay showed that RosA combined with ADM significantly induced DNA damage (TUNEL-positive staining) in the HepG2 and Bel-7402 groups (p<0.05). RosA combined with ADM significantly reduced Bcl-2 expression in HepG2 or Bel-7402 cells (p<0.05). RosA combined with ADM significantly increased Bax expression in HepG2 and Bel-7402 cells (p<0.05). Cell viability, apoptosis, cell cycle, and Bcl-2 and Bax expression were changed with increased concentrations of RosA. CONCLUSIONS RosA combined with ADM damaged tumor cell morphologies, decreased cell viability, and induced apoptosis of HepG2 and Bel-7402 by triggering the mitochondria-mediated signaling pathway.

Topics: Apoptosis; bcl-2-Associated X Protein; Carcinoma, Hepatocellular; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cinnamates; Depsides; Doxorubicin; Hep G2 Cells; Humans; Liver Neoplasms; Mitochondria; Proto-Oncogene Proteins c-bcl-2; Rosmarinic Acid; Signal Transduction

The aim of this study was to explore the anti-tumor effect and therapeutic potential of rosmarinic acid (RA) in the treatment of hepatocellular carcinoma (HCC). RA at 75, 150 and 300 mg/kg was given to H22 tumor-bearing mice by intragastric administration once daily for 10 consecutive days. Levels of inflammatory and angiogenic factors, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), and transforming growth factor-β (TGF-β) were measured by enzyme linked immunosorbent assays (ELISA). Protein levels of phosphorylated NF-κB p65 and p65 were detected by western blot. mRNA level of NF-κB p65 was analyzed by qRT-PCR. The results showed that RA could effectively suppress tumor growth with fewer toxic effects by regulating the secretion of cytokines associated with inflammation and angiogenesis, and suppressing the expression of NF-κB p65 in the xenograft microenvironment. Our findings unveil the possible anti-tumor mechanisms of RA and support RA as a potential drug for the treatment of HCC.

Topics: Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents, Non-Steroidal; Carcinoma, Hepatocellular; Cinnamates; Depsides; Inflammation; Inflammation Mediators; Liver Neoplasms, Experimental; Male; Mice; NF-kappa B; Rosmarinic Acid; Signal Transduction; Xenograft Model Antitumor Assays

Perilla frutescens (L.) Britt. (Lamiaceae) has traditionally been used to treat diseases, including tumors, but the antitumorigenesis mechanism is unclear. We evaluated the effects of Perilla frutescens leaf extract (PLE) on proliferation and apoptosis inducing in human hepatoma HepG2 cells using a cell proliferation assay, flow cytometry, and cDNA microarrays. Gene expression and apoptosis were also assessed in HepG2 cells treated with a major constituent of PLE, rosmarinic acid (RosA). In the PLE-treated HepG2 cells, antiproliferative activity (105 microg/mL) were observed, flow cytometry revealed significant apoptosis, and microarray data indicated that the expression of a lot apoptosis-related genes were regulated in a time-dependent manner. Compared with PLE, RosA (10 microg/mL; a dose equivalent to 105 microg/mL of PLE) was less effective in increasing the expression of apoptosis-related genes and apoptosis inducing in HepG2 cells. Thus, additional PLE constituents may influence apoptosis in HepG2 cells. The results of our study suggest that the PLE should be further investigated as a promising to treat hepatocellular carcinoma.

Topics: 4-1BB Ligand; Apoptosis; Caffeic Acids; Carcinoma, Hepatocellular; Caspase 8; Cell Line, Tumor; Cell Proliferation; Chromatography, High Pressure Liquid; Cinnamates; Cluster Analysis; Depsides; Dose-Response Relationship, Drug; Flow Cytometry; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; NF-kappaB-Inducing Kinase; Oligonucleotide Array Sequence Analysis; Perilla frutescens; Plant Extracts; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-jun; Reverse Transcriptase Polymerase Chain Reaction; Rosmarinic Acid; Time Factors