Mode of interaction between butyroyloxymethyl-diethyl phosphate (AN-7) and doxorubicin in MCF-7 and resistant MCF-7/Dx cell lines
Mode of interaction between butyroyloxymethyl-diethyl phosphate (AN-7) and doxorubicin in MCF-7 and resistant MCF-7/Dx cell lines
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DOI：
10.1007/s00432-006-0116-6
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发表时间：
2006-10-01
影响因子：
3.6
通讯作者：
Rephaeli, Ada
中科院分区：
文献类型：
--
作者：
Engel, Dikla;Nudelman, Abraham;Rephaeli, Ada
目的探讨丁酸和甲醛的前药磷酸丁酰氧基甲基二乙酯（AN-7）单用和联用对人肿瘤MCF-7和多重耐药MCF-7 Dx细胞株的抗癌活性和作用方式。方法采用Hoechst细胞活力法、菌落形成法以及碘化丙啶染色细胞和膜联蛋白V-FITC染色细胞FACS法检测AN-7单用或联用阿霉素的抗癌活性。Western blot检测蛋白表达和乙酰化的变化。用c -14标记的阿霉素来评价形成的阿霉素- dna加合物的数量。结果AN-7与同源前药对耐药和敏感细胞均表现出相似的生长抑制作用，其部分抑癌作用是通过抑制HDAC来实现的。AN-7瞬间增强组蛋白乙酰化，增加p21的表达。an -7和阿霉素的协同作用在敏感和耐药细胞系中得到证实，并通过FACS分析进一步证实，细胞死亡率增加。用c -14标记的阿霉素和AN-7处理的细胞分离的总基因组DNA中阿霉素-DNA加合物的数量比只用阿霉素处理的细胞显著增加。AN-7或阿霉素治疗增加了p53乙酰化，并在两者联合后进一步增强。结论AN-7联合阿霉素克服了耐药；至少部分原因是细胞内可释放的甲醛增加了阿霉素- dna加合物的形成，丁酸诱导组蛋白和p53乙酰化。由于多柔比星的使用受到毒性的限制，因此联合使用可能为乳腺癌提供一种有效的治疗方式，而且毒性较低。
Purpose To investigate the anticancer activity and mode of action of butyroyloxymethyl-diethyl phosphate (AN-7), a prodrug of butyric acid and formaldehyde, as a single agent and in combination with doxorubicin in human carcinoma MCF-7 and the multidrug resistant MCF-7 Dx cell lines.Methods The anti-cancer activity of AN-7 as a single agent or in combination with doxorubicin was measured by the Hoechst cell viability and colony forming assays as well as by FACS analyses of cells stained with propidium iodide and annexin V-FITC. Modulations of protein expression and acetylation were measured by Western blot analyses. The number of doxorubicin-DNA adducts formed was evaluated using C-14-labeled doxorubicin.Results The AN-7 and homologous prodrugs exhibited similar growth inhibition effects against drug resistant and sensitive cells, and elicited their anticancer effect partially by inhibition of HDAC. The AN-7 transiently augmented histone acetylation and increase of p21 expression. Synergy between AN-7 and doxorubicin was demonstrated in the sensitive and the resistant cell lines by viability and colony formation assays and was further confirmed by FACS analysis showing an increase in cell mortality. The number of doxorubicin-DNA adducts in total genomic DNA isolated from cells treated with C-14-labeled doxorubicin and AN-7 increased substantially compared to treatment with doxorubicin only. Treatment with AN-7 or doxorubicin increased p53 acetylation that was further potentiated by their combination.Conclusion The AN-7 combined with doxorubicin overcame drug resistance; at least in part by the intracellularly releasable formaldehyde that augmented formation of doxorubicin-DNA adducts and butyric acid that induced histone and p53 acetylation. Since the use of doxorubicin is limited by toxicity, the combination could offer an effective treatment modality with lower toxicity for breast cancer.