Long Liangyuan, Hu Xiangnan, Li Xiaoli, Zhou Duanfang, Shi Yun, Wang Lingen, Zeng Hongfang, Yu Xiaoping, Zhou Weiying
College of Pharmacy, Chongqing Medical University, Chongqing 400016, People's Republic of China.
Chongqing Key Laboratory of Drug Metabolism, Chongqing 400016, People's Republic of China.
Onco Targets Ther. 2020 May 4;13:3777-3788. doi: 10.2147/OTT.S240252. eCollection 2020.
Compound adduct is a eutectic crystal formed by non-covalent bonds of two compounds or multiple compounds with water. Emerging evidence suggests that adduct could be different from the simple physical mixture of the individual compounds and has some new features. Recent studies reported that both glimepiride (Gli) and metformin (Met) may possess an anti-breast cancer effect besides anti-diabetic effect. In the current study, we synthesized glimepiride-metformin adduct (GMA) and examined its anti-breast cancer effect in vitro and in vivo to explore its potential in treatment of breast cancer in diabetic patients.
GMA was synthesized from Gli, Met and water at a molar molecular mass of 1:1:1 and identified by infrared spectroscopy. MTT assay, colony formation assay and wound healing assay were performed to examine the effects of GMA on cell viability and migration of human breast cancer cell lines CAL-148, MDA-MB-453, MDA-MB-231and MCF-7. The effect of GMA on cell cycle and apoptosis was examined by flow cytometry. The orthotopic implantation model was established to observe the inhibitory effect of GMA on tumor growth. The expression of Ki67 was detected by immunohistochemistry. RT-qPCR and Western blotting were performed to investigate mechanisms for the function of GMA.
Both MTT and colony formation assays showed that GMA inhibited breast cancer cell viability, and the effect was greater than Gli alone, Met alone and the combination. In vivo study showed that GMA had an inhibitory effect on tumor growth of CAL-148 xenografts. Flow cytometry analysis indicated that GMA induced G1/S phase cell cycle arrest and apoptosis in breast cancer cells. RT-qPCR and Western blotting analyses showed that GMA activated AMPK, and up-regulated expression of p53 and p21, and down-regulated expression of cyclin D1 and CDK4.
GMA suppresses cell viability of breast cancer cells, and its effect is greater than Gli and Met alone or combination at the same concentration. GMA inhibits breast cancer cell growth in vivo. The antitumor effect of GMA may be related to the activation of AMPK resulting in up-regulation of p53 and p21 and down-regulation of cyclin D1 and CDK4.
复合加合物是由两种或多种化合物与水通过非共价键形成的低共熔晶体。新出现的证据表明，加合物可能不同于各化合物的简单物理混合物，具有一些新特性。最近的研究报道，格列美脲（Gli）和二甲双胍（Met）除具有抗糖尿病作用外，可能还具有抗乳腺癌作用。在本研究中，我们合成了格列美脲 - 二甲双胍加合物（GMA），并在体外和体内检测其抗乳腺癌作用，以探索其在糖尿病患者乳腺癌治疗中的潜力。
GMA由Gli、Met和水按摩尔分子质量1:1:1合成，并通过红外光谱进行鉴定。采用MTT法、集落形成试验和伤口愈合试验检测GMA对人乳腺癌细胞系CAL - 148、MDA - MB - 453、MDA - MB - 231和MCF - 7细胞活力和迁移的影响。通过流式细胞术检测GMA对细胞周期和凋亡的影响。建立原位植入模型以观察GMA对肿瘤生长的抑制作用。通过免疫组织化学检测Ki67的表达。进行RT - qPCR和蛋白质印迹分析以研究GMA发挥作用的机制。
MTT法和集落形成试验均显示，GMA抑制乳腺癌细胞活力，且该作用大于单独使用Gli、单独使用Met以及二者联合使用。体内研究表明，GMA对CAL - 148异种移植瘤的肿瘤生长具有抑制作用。流式细胞术分析表明，GMA诱导乳腺癌细胞G1/S期细胞周期阻滞和凋亡。RT - qPCR和蛋白质印迹分析表明，GMA激活AMPK，上调p53和p21的表达，下调细胞周期蛋白D1和细胞周期蛋白依赖性激酶4（CDK4）的表达。
GMA抑制乳腺癌细胞活力，其作用大于相同浓度下单独使用Gli和Met或二者联合使用。GMA在体内抑制乳腺癌细胞生长。GMA的抗肿瘤作用可能与激活AMPK导致p53和p21上调以及细胞周期蛋白D1和CDK4下调有关。
