肉桂酰胺格尔德霉素的体内外抗肿瘤活性

Anticancer effect of 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin:in vitro and in vivo

探讨化合物肉桂酰胺格尔德霉素（CDG）的体内外抗肿瘤活性。采用MTT法检测肿瘤细胞增殖。免疫荧光法和Annexin V-FITC／PI双染法检测肿瘤细胞凋亡，Transwell法检测肿瘤细胞迁移能力，Western blotting法检测肿瘤细胞中RAF．1、EGFR、AKT、CDK4、HER．2蛋白水平。使用异种移植瘤裸鼠模型检测CDG对人乳腺癌MCF-7细胞的生长抑制作用。结果表明：CDG对不同肿瘤细胞增殖具有抑制作用，IC50为13．6～67．4btg·mL-1；CDG能够诱导肿瘤细胞凋亡，引起多种细胞形态学改变；同时，CDG还可降低肿瘤细胞迁移能力，减少RAF-1、EGFR、AKT、CDK4、HER．2蛋白水平；体内实验结果显示，CDG毒性明显低于格尔德霉素（GDM），并且能够抑制人乳腺癌MCF-7裸鼠移植瘤生长。

In the present study, a new compound named 17-（6-cinnamamido-hexylamino-）-17-de- methoxygeldanamycin （CDG） was obtained by introducing the cinnamic acid （CA） group into the 17-site of geldanamycin （GDM）. The anti-cancer effects of CDG in vitro and in rive were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in rive. The results showed that CDG inhibited the proliferation of cancer cells （ICs0：13.6-67.4 }~g.mL-1）. After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 }~g.mL-l CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CD（3. The decreased levels of RAF- 1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.