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The role of cyclooxygenase‐2 in berberine induced apoptosis and inhibited cell migration of human gastric adenocarcinoma RF‐1 and RF‐48 cell lines
Author(s) -
YU CHUNSHU,
Kuo HsiuMaan,
Chung JingGung
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1131-d
Subject(s) - berberine , apoptosis , blot , matrix metalloproteinase , chemistry , flow cytometry , cell culture , cell , cancer cell , microbiology and biotechnology , cancer research , cancer , biology , biochemistry , medicine , gene , genetics
Berberine had been demonstrated to induce apoptosis in human many cancer cells but the role of cyclooxygenase‐1 and ‐2 (COX‐1 and ‐2) in berberine affecting cell migration of human gastric adenocarcinoma RF‐1 and RF‐48 cell lines was not reported. Compounds inhibiting COX‐2 transcriptional activity have therefore potentially a chemopreventive property against cancer cells. Both RF‐1 and RF‐48 cells were treated with berberine, and the apoptosis was measured by flow cytometry for examining of caspase‐3 activity. The expression of COX‐2, and matrix metalloproteinases in berberine‐treated RF‐1 and RF‐48 cells were assessed by Western blots. The results indicated that berberine induced apoptosis in both cells examined and berberine treatment inhibited COX‐2 and MMP‐2 and ‐9 expression dose‐dependently, but not COX‐1 and MMP‐7. An assay method for estimating COX‐2 transcriptional activity in both cell lines was established using a beta‐galactosidase reporter gene system, and examination was made of various concentrations of berberine for an inhibitory effect on COX‐2 transcriptional activity. We found that berberine effectively inhibits COX‐2 transcriptional activity, levels of mRNA and protein in both examined cancer cells in a dose‐ and time‐dependent manner. We observed that berberine affects levels of matrix metalloproteinase‐2 and ‐9 and the gene expression of NF‐κB/ p65 in a time course‐dependent but did not affect NF‐κB/ p50 based on the Western blotting and polymerase chain reaction methods.