A novel matrine derivate inhibits differentiated human hepatoma cells and hepatic cancer stem-like cells by suppressing PI3K/AKT signaling pathways  

作　　者：Ying LIU Yang QI Zhi-hui BAI Chen-xu NI Qi-hui REN Wei-heng XU Jing XU Hong-gang HU Lei QIU Jian-zhong LI Zhi-gao HE Jun-ping ZHANG 

机构地区：[1]Department of Pharmacy, Shanghai East Hospital, Tongji University, Shanghai 310000, China [2]College of Pharmacy, Second MilitaryMedical University, Shanghai 200433, China

出　　处：《Acta Pharmacologica Sinica》2017年第1期120-132,共13页中国药理学报（英文版）

基　　金：Acknowledgements This project was supported by grants from the National Natural Science Foundation of China （No 81270508） and the National Major Special Science and Technology Project （No 2012ZX09103101-043）. The funding agency had no role in the actual experimental design, analysis, or writing of this manuscript.

摘　　要：Matrine is an alkaloid extracted from a Chinese herb Sophora flavescens Ait, which has shown chemopreventive potential against various cancers. In this study, we evaluated the anticancer efficacy of a novel derivative of matrine, （6aS, 10S, 11aR, 11bR, 11cS）- lO-methylamino-dodecahydro-3a,7a-diazabenzo （de） anthracene-8-thione （MASM）, against human hepatocellular carcinoma （HCC） ceils and their corresponding sphere cells in vitro and in vivo. Human HCC cell lines （Hep3B and Huh7） were treated with MASM. Cell proliferation was assessed using CCK8 and colony assays; cell apoptosis and cel{ cycle distributions were examined with flow cytometry. The expression of cell markers and signaling molecules was detected using Western blot and qRT-PCR analyses. A sphere culture technique was used to enrich cancer stem cells （CSC） in Hep3B and Huh7 cells. The in vivo antitumor efficacy of MASM was evaluated in Huh7 cell xenograft model in BALB/c nude mice, which were administered MASM （10 mg.kg-^-d~, ig） for 3 weeks. After the treatment was completed, tumor were excised and weighed. A portion of tumor tissue was enzymatically dissociated to obtain a single cell suspension for the spheroid formation assays. MASM （2, 10, 20 pmol/L） dose-dependently iahibited the proliferation of HCC cells, and induced apoptosis, which correlated with a reduction in Bcl-2 expression and an increase in PARP cleavage. MASM also induced cell cycle arrest in Go/G~ phase, which was accompanied by increased p27 and decreased Cyclin D1 expression, interestingly, MASM （2, 10, and 20 pmoVL） drastically reduced the EpCAM＋/CD133＊ cell numbers, suppressed the sphere formation, inhibited the expression of stem cell marker genes and promoted the expression of mature hepatocyte markers in the Hep3B and Huh7 spheroids. Additionally, MASM dose-dependently suppressed the PI3K/AKT/mTOR and AKT/GSK313/13-catenin signaling pathways in Hep3B and Huh7 cells. In Huh7 xenograft bearing nude mice, MASM administration significantly iMatrine is an alkaloid extracted from a Chinese herb Sophora flavescens Ait, which has shown chemopreventive potential against various cancers. In this study, we evaluated the anticancer efficacy of a novel derivative of matrine, （6aS, 10S, 11aR, 11bR, 11cS）- lO-methylamino-dodecahydro-3a,7a-diazabenzo （de） anthracene-8-thione （MASM）, against human hepatocellular carcinoma （HCC） ceils and their corresponding sphere cells in vitro and in vivo. Human HCC cell lines （Hep3B and Huh7） were treated with MASM. Cell proliferation was assessed using CCK8 and colony assays; cell apoptosis and cel{ cycle distributions were examined with flow cytometry. The expression of cell markers and signaling molecules was detected using Western blot and qRT-PCR analyses. A sphere culture technique was used to enrich cancer stem cells （CSC） in Hep3B and Huh7 cells. The in vivo antitumor efficacy of MASM was evaluated in Huh7 cell xenograft model in BALB/c nude mice, which were administered MASM （10 mg.kg-^-d~, ig） for 3 weeks. After the treatment was completed, tumor were excised and weighed. A portion of tumor tissue was enzymatically dissociated to obtain a single cell suspension for the spheroid formation assays. MASM （2, 10, 20 pmol/L） dose-dependently iahibited the proliferation of HCC cells, and induced apoptosis, which correlated with a reduction in Bcl-2 expression and an increase in PARP cleavage. MASM also induced cell cycle arrest in Go/G~ phase, which was accompanied by increased p27 and decreased Cyclin D1 expression, interestingly, MASM （2, 10, and 20 pmoVL） drastically reduced the EpCAM＋/CD133＊ cell numbers, suppressed the sphere formation, inhibited the expression of stem cell marker genes and promoted the expression of mature hepatocyte markers in the Hep3B and Huh7 spheroids. Additionally, MASM dose-dependently suppressed the PI3K/AKT/mTOR and AKT/GSK313/13-catenin signaling pathways in Hep3B and Huh7 cells. In Huh7 xenograft bearing nude mice, MASM administration significantly i

关 键 词：MATRINE MASM hepatoceliular carcinoma Hep3B cells Huh7 cells cancer stem cells xenograft nude mouse model PI3K/AKT pathway 

分 类 号：Q813[生物学—生物工程] Q691.5