Prostanoid,EP1 receptor as the target of （-）-epigallocatechin-3-gallate in suppressing hepatocellular carcinoma cells in vitro  

作　　者：Juan JIN Yan CHANG Wei WEI Yi-fu HE Shan-shan HU Di WANG Yu-jing WU 

机构地区：[1]Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immunopharmacology of the Education Ministry of China,Hefei 230032,China

出　　处：《Acta Pharmacologica Sinica》2012年第5期701-709,共9页中国药理学报（英文版）

基　　金：Acknowledgements This work was supported by the National Natural Science Foundation of China （No 30973543 and No 30572356）, and by the Scientific Research Foundation of Anhui Medical University （No 2008kj13）. The authors acknowledge ONO Pharmaceutical Co in Japan for providing the selective EP1 receptor agonist and antagonist. The authors especially thank Yuri SHEIKINE for rearranging and correcting the manuscript.

摘　　要：Aim： To investigate the effects of （-）-epigallocatechin-3-gallate （EGCG）, an active compound in green tea, on prostaglandin E2 （PGE2）- induced proliferation and migration, and the expression of prostanoid EP1 receptors in hepatocellular carcinoma （HCC） cells. Methods： HCC cell line HepG2, human hepatoma cell lines MHCC-97L, MHCC-97H and human hepatocyte cell line L02 were used. Cell viability was analyzed using MTr assay. PGE2 production was determined with immunoassay. Wound healing assay and transwel filter assay were employed to assess the extent of HCC cell migration. The expression of EP1 receptor and Gq protein were examined using Western blot assay. Results： PGE2 （4-40000 nmol/L） or the EP1 receptor agonist ONO-DI-O04 （400-4000 nmol/L） increased the viability and migration of HepG2 cells in concentration-dependent manners. EGCG （100 pg/mL） significantly inhibited the viability and migration of HepG2 cells induced by PGE2 or ONO-DI-O04. HepG2 cells secreted an abundant amount of PGE2 into the medium, and EGCG （100 pg/mL） significantly inhibited the PGE2 production and EP1 receptor expression in HepG2 cells. EGCG （100 pg/mL） also inhibited the viability of MHCC-97L cells, but not that of MHCC-97H cells. Both EGCG （100 pg/mL） and EP1 receptor antagonist 0N0-8711 inhibited PGE2 4 pmol/L and ONO-DI-O04 400 nmol/L-induced growth and migration of HepG2 cells. Both EGCG （100 pg/mL） and 0N0-8711 210 nmol/L inhibited PGE2- and ONO-Dl-OO4-induced EP~ expression. EGCG and 0N0-8711 had synergistic effects in inhibiting EP1 receptor expression. PGE2, ONO-DI-004, 0N0-8711, and EGCG had no effects on Gq expression in HepG2 cells, respectively. Conclusion： These findings suggest that the anti-HCC effects of EGCG might be mediated, at least partially, through the suppressing EP1 receptor expression and PGE2 production.Aim： To investigate the effects of （-）-epigallocatechin-3-gallate （EGCG）, an active compound in green tea, on prostaglandin E2 （PGE2）- induced proliferation and migration, and the expression of prostanoid EP1 receptors in hepatocellular carcinoma （HCC） cells. Methods： HCC cell line HepG2, human hepatoma cell lines MHCC-97L, MHCC-97H and human hepatocyte cell line L02 were used. Cell viability was analyzed using MTr assay. PGE2 production was determined with immunoassay. Wound healing assay and transwel filter assay were employed to assess the extent of HCC cell migration. The expression of EP1 receptor and Gq protein were examined using Western blot assay. Results： PGE2 （4-40000 nmol/L） or the EP1 receptor agonist ONO-DI-O04 （400-4000 nmol/L） increased the viability and migration of HepG2 cells in concentration-dependent manners. EGCG （100 pg/mL） significantly inhibited the viability and migration of HepG2 cells induced by PGE2 or ONO-DI-O04. HepG2 cells secreted an abundant amount of PGE2 into the medium, and EGCG （100 pg/mL） significantly inhibited the PGE2 production and EP1 receptor expression in HepG2 cells. EGCG （100 pg/mL） also inhibited the viability of MHCC-97L cells, but not that of MHCC-97H cells. Both EGCG （100 pg/mL） and EP1 receptor antagonist 0N0-8711 inhibited PGE2 4 pmol/L and ONO-DI-O04 400 nmol/L-induced growth and migration of HepG2 cells. Both EGCG （100 pg/mL） and 0N0-8711 210 nmol/L inhibited PGE2- and ONO-Dl-OO4-induced EP~ expression. EGCG and 0N0-8711 had synergistic effects in inhibiting EP1 receptor expression. PGE2, ONO-DI-004, 0N0-8711, and EGCG had no effects on Gq expression in HepG2 cells, respectively. Conclusion： These findings suggest that the anti-HCC effects of EGCG might be mediated, at least partially, through the suppressing EP1 receptor expression and PGE2 production.

关 键 词：hepatocellular carcinoma epigallocatechin-3-gallate prostaglandin E2 prostanoid EP1 receptor 

分 类 号：Q593.2[生物学—生物化学] TS252.1[轻工技术与工程—农产品加工及贮藏工程]