Anti-invasion Effect of Crebanine and <i>O</i>-Methylbulbocapnine from <i>Stephania venosa</i> <i>via</i> Down-Regulated Matrix Metalloproteinases and Urokinase Plasminogen Activator

Abstract

The alkaloids isolated from Stephania venosa (S. venosa) have been shown to inhibit the proliferation and to induce the apoptosis of cancer cells. However, the anti-metastatic effect of the alkaloids on cancer cell invasion is unknown. In this study, we investigated the anti-invasive properties of four alkaloids from S. venosa, crebanine (CN), O-methylbulbocapnine (OMBC), tetrahydropalmatine (THP), and N-methyltetrahydropalmatine (NMTHP), in HT1080 human fibrosacroma cells. Treatment of the cells with 15 µg/mL of CN and OMBC reduced the chemo-invasion of HT1080 cells to 45 and 50%, respectively, whereas THP and NMTHP had a negative effect. On the other hand, CN and OMBC had no effect on cell migration. Matrix metalloproteinases (MMPs) and urokinase plasminogen activator (uPA) are the extracellular matrix (ECM) degradation enzymes that play an important role in cancer cell metastasis. Results from zymography and western blot analysis showed that CN and OMBC comparatively reduced MMP-2, MMP-9, MT1-MMP and uPA expression in a dose-dependent manner. However, CN and OMBC had no effect on the activity of collagenase, MMP-2 and MMP-9. We also found that CN and OMBC reduced the nuclear translocation and DNA binding activity of nuclear factor kappa B (NF-κB), which is the expressed mediator of ECM degradation enzymes. These findings demonstrated that CN and OMBC mediated HT1080 cell invasion by the reduction of MMP-2, MMP-9, uPA and MT1-MMP expression, possibly by targeting of NF-κB signaling pathway in the HT1080 cells.

Proanthocyanidin in Red Rice Inhibits MDA-MB-231 Breast Cancer Cell Invasion <i>via</i> the Expression Control of Invasive Proteins

“…13) Briefly, MDA-MB-231 cells were treated with various concentrations of PRFR (0-20 µg/mL) for 24 h in DMEM serum-free medium. The culture supernatant was collected and separated by 10% polyacrylamide gels containing 0.1% w/v of gelatin in a non-reducing condition.…”

“…13) Briefly, MDA-MB-231 cells were treated with various concentrations of PRFR for 24 h. The treated cells were washed with ice clod phosphate buffered saline (PBS) and collected by the cell scraper. The cells was lysated with a lysis buffer containing a proteinase inhibitor (50 mM Tris-HCl, 150 mM NaCl, 10 mM, EDTA, 1% Triton X-100, 1 mM phenylmethylsulfonyl fluoride (PMSF), 10 µg/mL leupeptin, 10 µg/ mL aprotinin) for 20 min on ice.…”

“…The DNA-binding activity of NF-κB was evaluated using NF-κB (p65) Transcription Factor Assay Kit (Cayman Company, Ann Arbor, MI, U.S.A.) with slight modifications. 13) The nuclear proteins at 10 µg/mL were added to the wellcoated double-stranded DNA (dsDNA) templates carrying NF-κB response elements and incubated at 4°C overnight. Blank wells, positive control (provided with kit) and nonspecific binding samples were also incubated on the plate.…”

Alkaloids from <i>Stephania venosa</i> as Chemo-Sensitizers in SKOV3 Ovarian Cancer Cells <i>via</i> Akt/NF-κB Signaling

“…In recent years, CN has gained much attention due to its biological activities and proven pharmacological safety. 7,8) It was shown that CN more selectively decrease the survival and proliferation of cancerous cell lines than it does normal human fibroblasts.…”

“…In recent years, CN has gained much attention due to its biological activities and proven pharmacological safety. 7,8) It was shown that CN more selectively decrease the survival and proliferation of cancerous cell lines than it does normal human fibroblasts.9) Moreover, CN has anti-invasive property in cancer cells by inhibiting constitutive nuclear factor kappa B (NF-κB) activation.10) On the other hand, THP has been stated to possess multi-drug resistant reversing properties in human breast cancer cells.11) These established results have led us to hypothesize that these natural alkaloids may possibly improve the effectiveness of the traditional anticancer drugs.Although the anti-cancer properties of CN have been studied in numerous cancer cell lines, it remains unidentified whether CN has chemo-sensitizing effect on drug-resistant cancer cells. Therefore, the purpose of the present study was to determine whether CN and its natural analogues show chemo-sensitizing effects in response to platinum drug in human…”