University of Pretoria, Faculty of Health Sciences, Department of Physiology, Pretoria 0007, South Africa.
University of Zagreb, Faculty of Pharmacy and Biochemistry, HR-10 000 Zagreb, Croatia.
Acta Pharm. 2018 Sep 1;68(3):337-348. doi: 10.2478/acph-2018-0021.
In our previous paper, we showed that three primaquine-cinnamic acid conjugates composed of primaquine (PQ) residue and cinnamic acid derivatives (CADs) bound directly by an amide linkage (1) or through an acylsemicarbazide spacer (2 and 3) had significant growth inhibitory effects on some cancer cell lines. Compound 1 induced significant growth inhibition in the colorectal adenocarcinoma (SW620), human breast adenocarcinoma (MCF-7) and cervical carcinoma (HeLa) cell lines, while compounds 2 and 3 selectively inhibited the growth of MCF-7 cells. To better understand the underlying mechanisms of action of these PQ-CADs, morphological studies of the effects of test compounds on MCF-7 cells were undertaken using haematoxylin and eosin stain. Further analysis to determine the effects of test compounds on caspase activity and on the levels of apoptosis proteins were undertaken using the enzyme-linked immunosorbent assay (ELISA). Haematoxylin and eosin staining revealed that compounds 1 and 3 induced morphological changes in MCF-7 cells characteristic of apoptosis, while 2-treated cells were in interphase. Cell cycle analysis showed that cells treated with 1 and 3 were in sub-G1, while cells treated with 2 were mainly in interphase (G1 phase). Further, the study showed that the treatment of MCF-7 cells with 1 and 3 resulted in poly ADP ribose polymerase (PARP) cleavage as well as caspase-9 activation, indicating that they induced apoptotic cell death. We further investigated their effects on two important processes during metastasis, namely, migration and invasion. Compounds 1 and 3 inhibited the migration and invasion of MCF-7 cells, while compound 2 had a marginal effect.
在我们之前的论文中，我们表明三种由伯氨喹（PQ）残基和肉桂酸衍生物（CAD）组成的伯氨喹-肉桂酸缀合物通过酰胺键（1）或通过酰基缩氨基脲间隔基（2 和 3）直接结合，对一些癌细胞系具有显著的生长抑制作用。化合物 1 对结直肠腺癌（SW620）、人乳腺癌（MCF-7）和宫颈癌（HeLa）细胞系诱导了显著的生长抑制，而化合物 2 和 3 则选择性地抑制 MCF-7 细胞的生长。为了更好地了解这些 PQ-CAD 的作用机制，我们使用苏木精和伊红染色对测试化合物对 MCF-7 细胞的影响进行了形态学研究。进一步分析以确定测试化合物对 Caspase 活性和凋亡蛋白水平的影响，使用酶联免疫吸附测定（ELISA）进行。苏木精和伊红染色显示，化合物 1 和 3 诱导 MCF-7 细胞发生特征性凋亡的形态变化，而 2 处理的细胞处于间期中。细胞周期分析显示，用 1 和 3 处理的细胞处于 sub-G1 期，而用 2 处理的细胞主要处于间期中（G1 期）。此外，研究表明，用 1 和 3 处理 MCF-7 细胞导致多聚 ADP 核糖聚合酶（PARP）切割以及 caspase-9 激活，表明它们诱导了细胞凋亡。我们进一步研究了它们对转移过程中两个重要过程，即迁移和侵袭的影响。化合物 1 和 3 抑制 MCF-7 细胞的迁移和侵袭，而化合物 2 则仅有轻微作用。
