Mousavi Elham, Tavakolfar Shahrzad, Almasirad Ali, Kooshafar Zahra, Dehghani Soudeh, Afsharinasab Ahoo, Amanzadeh Amir, Shafiee Samira, Salimi Mona
Department of Biology, Science and Research Branch Islamic Azad University, Tehran, Iran.
Department of Medicinal Chemistry, Faculty of Pharmacy, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
Cancer Chemother Pharmacol. 2017 Jun;79(6):1195-1203. doi: 10.1007/s00280-017-3318-5. Epub 2017 Apr 27.
The hydrazide backbone is a well-known structural core system found in a broad range of biologically activated compounds. Among which, the compounds with anticancer activity have been cited in a number of studies. With this object in mind, we focused on the in vitro and in vivo anticancer potential of two novel hydrazide derivatives bearing furan or thiophen substituents (compounds 1 and 2).
The cytotoxic property was evaluated using MTT assay against MCF-7 human breast adenocarcinoma cell line, while the in vivo antitumor activity was investigated in BALB/c mice bearing 4T1 mammary carcinoma cells. Flow cytometry was used for cell cycle analysis, and detection of apoptosis was examined by Annexin-V-FLUOS/PI assay. Protein expression of Bax, Bcl-2 and procaspase-3 was estimated by Western blotting.
Compounds 1 and 2 were found to be cytotoxic towards breast cancer cells presenting IC values of 0.7 and 0.18 µM, respectively, and selectivity over normal fibroblast cells. Our findings further indicated that 2 × IC concentrations of both compounds induce early stage apoptosis and increase percentage of sub-G1 population in MCF-7 cells at 48 h. An elevation in Bax/Bcl-2 ratio and caspase-3 cleavage suggested that apoptosis induced by the two compounds is through a caspase- and mitochondrial-dependent pathway. In the in vivo study, compounds 1 and 2 at doses of 10 and 1 mg/Kg/day, respectively, significantly hindered the growth of tumor after 3 weeks of i.p. administration, when compared to vehicle-treated mice.
Collectively, the great potential exhibited by compound 2 could make it a promising chemotherapeutic candidate for human cancers, especially for breast cancer.
酰肼主链是在多种生物活性化合物中发现的一种著名的结构核心体系。其中，具有抗癌活性的化合物已在多项研究中被提及。基于此目的，我们聚焦于两种带有呋喃或噻吩取代基的新型酰肼衍生物（化合物1和2）的体外和体内抗癌潜力。
使用MTT法评估对MCF-7人乳腺腺癌细胞系的细胞毒性，同时在携带4T1乳腺癌细胞的BALB/c小鼠中研究体内抗肿瘤活性。采用流式细胞术进行细胞周期分析，并通过Annexin-V-FLUOS/PI法检测细胞凋亡。通过蛋白质印迹法估计Bax、Bcl-2和procaspase-3的蛋白表达。
发现化合物1和2对乳腺癌细胞具有细胞毒性，IC值分别为0.7和0.18 μM，且对正常成纤维细胞具有选择性。我们的研究结果进一步表明，两种化合物的2×IC浓度在48小时时可诱导MCF-7细胞早期凋亡并增加亚G1期细胞群体百分比。Bax/Bcl-2比值升高和caspase-3裂解表明这两种化合物诱导的凋亡是通过caspase和线粒体依赖性途径。在体内研究中，与溶剂处理的小鼠相比，化合物1和2分别以10和1 mg/Kg/天的剂量腹腔注射给药3周后，显著抑制了肿瘤生长。
总体而言，化合物2展现出的巨大潜力使其有望成为治疗人类癌症，尤其是乳腺癌的化疗候选药物。
