Mi Jin-xia, Wang Guang-feng, Wang Heng-bang, Sun Xiao-qing, Ni Xin-yan, Zhang Xiong-wen, Tang Jia-ming, Yang Da-jun
Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Acta Pharmacol Sin. 2008 Dec;29(12):1467-77. doi: 10.1111/j.1745-7254.2008.00901.x.
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).
The IC50 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.
The IC50 of ApoG2 in HCC cells was 17.28-30.63 micromol/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-XL, 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.
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.
研究去甲棉酚（ApoG2）单独或与阿霉素（ADM）联合应用对人肝细胞癌（HCC）的体外和体内活性及相关机制。
采用WST法检测ApoG2的体外半数抑制浓度（IC50），并用CalcuSyn法分析协同作用。使用4',6-二脒基-2-苯基吲哚染色和流式细胞术分析测定细胞凋亡。采用蛋白质免疫印迹法检测凋亡相关蛋白的表达。在裸鼠异种移植模型中评估体内活性，通过末端脱氧核苷酸转移酶介导的地高辛-dUTP缺口末端标记（TUNEL）法测定肿瘤组织中的凋亡情况。
ApoG2在肝癌细胞中的IC50为17.28 - 30.63微摩尔/升。当ApoG2与ADM联合使用时，与单独使用ApoG2相比，SMMC - 7721细胞的细胞毒性和凋亡增加。蛋白质免疫印迹结果表明，ApoG2通过下调抗凋亡蛋白Bcl - 2、Mcl - 1和Bcl - XL，上调促凋亡蛋白Noxa，并促进半胱天冬酶-9和-3的活性，诱导SMMC - 7721细胞凋亡。口服ApoG2可抑制裸鼠体内异种移植的SMMC - 7721肿瘤生长，且对正常组织无损害。体内研究还表明，当ApoG2以100或200毫克/千克剂量与5.5毫克/千克的ADM联合使用时，抗肿瘤作用增强，相对肿瘤增殖率（T/C）值分别为0.456和0.323。TUNEL法证实了ApoG2单独或与ADM联合在体内诱导肿瘤组织凋亡。
ApoG2是一种潜在的无毒靶向药物，通过上调Noxa诱导凋亡，同时抑制抗凋亡蛋白，并增强化疗药物ADM对肝癌的作用。
