Induction of apoptosis of human gastric carcinoma SGC-7901 cell line by 5, 7-dihydroxy-8-nitrochrysin in vitro.
OBJECTIVE
To investigate the effect of 5, 7-dihydroxy-8-nitrochrysin (NOChR) on apoptosis of human gastric carcinoma SGC-7901 cell line.


METHODS
SGC-7901 cells were cultured in vitro and the inhibitory effect of NOChR on proliferation of SGC-7901 cells was measured by using an MTT assay. NOChR-induced apoptosis rate of SGC-7901 cells was detected using flow cytometry (FCM) with PI staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of NOChR on the proxisome proliferator-activated receptor-gamma (PPARgamma), Bcl-2 and Bax protein expression of SGC-7901 cells was analyzed by Western blot.


RESULTS
MTT assay showed that NOChR markedly inhibited proliferation of SGC-7901 cells in a dose-dependent manner, and when IC(50) was 4.14 micromol/L, the potency of NOChR was 10 times than that of lead compound, chrysin (ChR, IC(50) was 40.56 micromol/L), and was similar to 5-fluorouracil (5-FU, IC(50) was 4.51 micromol/L). FCM with propidium iodide (PI) staining demonstrated that the apoptosis rates of SGC-7901 cells treated with 1.25, 5.00 and 20.00 micromol/L NOChR for 48 h were 9.8% +/- 0.2%, 36.8% +/- 1.9% and 45.5% +/- 3.5%, respectively, and were significantly higher when treated with 5.00 and 20.00 micromol/L NOChR than that with 20.00 micromol/L ChR (12.9% +/- 1.5%). DNA agarose gel electrophoresis showed that treatment of SGC-7901 cells with 20.00 micromol/L NOChR for 48 h resulted in typical DNA ladder bands of DNA of SGC-7901 cells, which could be eliminated by treating with 10.00 micromol/L GW9662, a blocker of PPARgamma. Western blot analysis revealed that after 24 h of treatment with 20.00 micromol/L NOChR, PPARgamma and Bax protein expression of SGC-7901 cells increased but Bcl-2 expression decreased; however, pre-incubation with 10.00 micromol/L GW9662 could efficiently antagonize and weaken the regulatory effect of 20.00 micromol/L NOChR on Bax and Bcl-2 protein expression of SGC-7901 cells.
Ai
Xiao-Hong
XH
Department of Oncology, First Affiliated Hospital, Nahua University, Hengyang 421001, Hunan Province, China.
Zheng
Xing
X
Tang
Xiao-Qing
XQ
Sun
Li
L
Zhang
Yang-Qin
YQ
Qin
Yong
Y
Liu
Hua-Qing
HQ
Xia
Hong
H
Cao
Jian-Guo
JG
D000813:Anilides; D017209:Apoptosis / Q000187:drug effects*; D002277:Carcinoma / Q000188:drug therapy* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004587:Electrophoresis, Agar Gel; D005419:Flavonoids / Q000494:pharmacology / Q000627:therapeutic use*; D005434:Flow Cytometry; D006801:Humans; D009574:Nitro Compounds / Q000494:pharmacology / Q000627:therapeutic use*; D047495:PPAR gamma / Q000378:metabolism*; D011419:Propidium; D014176:Protein Biosynthesis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D013778:Tetrazolium Salts; D013844:Thiazoles; D051028:bcl-2-Associated X Protein / Q000378:metabolism
C457499:2-chloro-5-nitrobenzanilide; C524514:5,7-dihydroxy-8-nitrochrysin; D000813:Anilides; D005419:Flavonoids; D009574:Nitro Compounds; D047495:PPAR gamma; D019253:Proto-Oncogene Proteins c-bcl-2; D013778:Tetrazolium Salts; D013844:Thiazoles; D051028:bcl-2-Associated X Protein; D011419:Propidium; C022616:thiazolyl blue