Anti-proliferative effect of Ginsenoside-Rg18 and panaxydol derived from Panax ginseng in human A549 lung cancer cells

다국어 초록 (Multilingual Abstract)

Although ginsenoside-Rg18, a novel dammarane-type triterpene saponin, and panaxydol, a polyacetylenic compound, derived from Panax ginseng roots have been reported to inhibit the growth of cancer cells, the underlying molecular mechanisms for the anti-proliferative effects of these compounds in the A549 non-small cell lung cancer (NSCLC) cells were not determined. Treatment of A549 cells with ginsenoside-Rg18 or panaxydol induced cell cycle arrest at G1 phase. These cell cycle arrest were accompanied by down-regulation of the, cyclin-dependent kinase/cyclin (CDK)2, CDK4, CDK6, cyclin D1, cyclin D2 and cyclin E and up-regulation of cyclin-dependent kinase inhibitor p21CIP1/WAF1 and p27KIP1 involved in G1 arrest, and decrease in the phosphorylated form of retinoblastoma protein. ginsenoside-Rg18 treatment caused intracellular accumulation of reactive oxygen species (ROS), whereas it dosedependently inhibited p38, JNK and p65 phosphorylation. However, panaxydol increased NADPH oxidase (NOX) -dependent intracellular ROS and suppressed the migration of A549 cells, which correlated with the inhibition of metalloproteinase-2 (MMP-2) and MMP-9 protein expression via inhibition of MAPK/AP-1 and NF-κB signaling. These results demonstrate that the anti-proliferative effects of ginsenosideRg18 and panaxydol are closely linked to the regulation of G1 arrest in human A549 lung cancer cells.

목차 (Table of Contents)

• I. INTRODUCTION 1
• II. MATERIAL AND METHODS 5
• 2.1 Reagents 5
• 2.2 Cell culture 5
• 2.3 Cytotoxicity test 6
• 2.4 Cell cycle analysis 6
• 2.5 Protein extraction and Western blot analysis 6
• 2.6 ROS detection (DCFH-DA assay) 7
• 2.7 Cell migration assay 7
• 2.8 Statistical analysis 8
• III. RESULTS & DISCUSSION 9
• 3.1 Ginsenoside-Rg18 from Panax ginseng root inhibits cell proliferation via induction of G1 arrest in lung cancer A549 cells 9
• 3.1.1 Ginsenoside Rg18 inhibited cell proliferation and induced G1 arrest in A549 cells 9
• 3.1.2 Effect of ginsenoside-Rg18 on protein levels of various G1 arrest regulators in A549 cells 13
• 3.1.3 Ginsenoside-Rg18 upregulated the intracellular ROS levels and downregulated p38, JNK and NF-κB activation in A549 cells 17
• 3.2 Panaxydol induces G1 arrest through p53 activation, NADPH oxidase-dependent generation of reactive oxygen species 22
• 3.2.1 Panaxydol induced G1 arrest in A549 cells 22
• 3.2.2 Effects of panaxydol on G1 arrest associated protein and p53 activation of A549 cells 24
• 3.2.3 NADPH oxidase dependent-oxidative stress mediates panaxydol-induced G1 arrest 26
• 3.3 Panaxydol inhibits A549 cell migration via inhibition of AP-1- and NF-κB-dependent matrix metalloproteinase-2 and -9 expression 29
• 3.3.1 Panaxydol inhibited A549 cell migration, MMP-2 and MMP-9 expression 29
• 3.3.2 Panaxydol inhibited NF-κB, MAPK and AP-1 activation 31
• IV. REFERENCES 33