Open AccessSynergistic antitumoral activity and induction of apoptosis by novel pan Bcl‐2 proteins inhibitor apogossypolone with adriamycin in human hepatocellular carcinoma
Author(s) -
MI Jinxia,
WANG Guangfeng,
WANG Hengbang,
SUN Xiaoqing,
NI Xinyan,
ZHANG Xiongwen,
TANG Jiaming,
YANG Dajun
Publication year - 2008
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2008.00901.x
Subject(s) - apoptosis , in vivo , tunel assay , chemistry , in vitro , blot , cancer research , hepatocellular carcinoma , pharmacology , biology , biochemistry , microbiology and biotechnology , gene
Aim: To investigate the in vitro and in vivo activities and related mechanism of apogossypolone (ApoG2) alone or in combination with adriamycin (ADM) against human hepatocellular carcinoma (HCC). Methods: The IC 50 of ApoG2 in vitro was tested by WST assay, and the synergistic effect was analyzed using the CalcuSyn method. Cell apoptosis was determined using 4′,6‐diamidino‐2‐phenylindole staining and flow cytometric analysis. Western blotting was used to determine the expression of apoptosis‐related proteins. In vivo activity was evaluated in the xenograft model in nude mice, and apoptosis in tumor tissues was determined by terminal deoxynucleotidyl transferase‐mediated digoxigenin‐dUTP nick‐end labeling (TUNEL) assay. Results: The IC 50 of ApoG2 in HCC cells was 17.28–30.63 μmol/L. When ApoG2 was combined with ADM, increased cytotoxicity and apoptosis were observed in SMMC‐7721 cells compared to treatment with ApoG2 alone. The Western blotting results indicated that the ApoG2 induced apoptosis in SMMC‐7721 cells by downregulating anti‐apoptotic proteins Bcl‐2, Mcl‐1, and Bcl‐X L , up‐regulating pro‐apoptotic protein Noxa, and promoting the activities of caspases‐9 and ‐3. The tumor growth of xenograft SMMC‐7721 was inhibited in nude mice when ApoG2 was administered orally without causing damage to the normal tissues. The in vivo study also indicated an increasing anti‐tumoral effect when ApoG2 at 100 or 200 mg/kg dosages were used together with ADM at 5.5 mg/kg, with relative tumor proliferation rate (T/C) values of 0.456 and 0.323, respectively. Apoptosis induced in vivo by ApoG2 alone or combined with ADM was confirmed by TUNEL assay in tumor tissues. Conclusion: ApoG2 is a potential non‐toxic target agent that induces apoptosis by upregulating Noxa, while inhibiting anti‐apoptotic proteins and promoting the effect of chemotherapy agent ADM in HCC.