Department of Pharmacology, School of Pharmacy, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Department of Chemistry, Laboratory of Organic Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Int J Oncol. 2018 Nov;53(5):2167-2179. doi: 10.3892/ijo.2018.4550. Epub 2018 Sep 3.
The present study aimed to assess the pharmacological anticancer profile of three natural and five synthetic sesquiterpenes developed by total chemical synthesis. To this end, their properties at the cellular and molecular level were evaluated in a panel of normal and cancer cell lines. The results obtained by performing cytotoxicity assays and gene expression analysis by reverse transcription-quantitative polymerase chain reaction showed that: i) Among the sesquiterpene derivatives analyzed, VDS58 exhibited a notable anticancer profile within attached (U-87 MG and MCF-7) and suspension (K562 and MEL-745) cancer cell cultures; however, U-87 MG cells were able to recover their proliferation capacity rapidly after 48 h of exposure; ii) gene expression profiling of U-87 MG cells, in contrast to K562 cells, showed a transient induction of cyclin-dependent kinase inhibitor 1A (CDKN1) expression; iii) the expression levels of transforming growth factor β1 (TGFB1) increased after 12 h of exposure of U-87 MG cells to VDS58 and were maintained at this level throughout the treatment period; iv) in K562 cells exposed to VDS58, TGFB1 expression levels were upregulated for 48 h and decrease afterwards; and v) the re-addition of VDS58 in U-87 MG cultures pretreated with VDS58 resulted in a notable increase in the expression of caspases (CASP3 and CASP9), BCL2‑associated agonist of cell death (BAD), cyclin D1, CDK6, CDKN1, MYC proto-oncogene bHLH transcription factor (MYC), TGFB1 and tumor suppressor protein p53. This upregulation persisted only for 24 h for the majority of genes, as afterwards, only the expression of TGFB1 and MYC was maintained at high levels. Through bioinformatic pathway analysis of RNA-Seq data of parental U-87 MG and K562 cells, substantial variation was reported in the expression profiles of the genes involved in the regulation of the cell cycle. This was associated with the differential pharmacological profiles observed in the same cells exposed to VDS58. Overall, the data presented in this study provide novel insights into the molecular mechanisms of action of sesquiterpene derivatives by dysregulating the expression levels of genes associated with the cell cycle of cancer cells.
本研究旨在评估三种天然和五种全合成倍半萜的药理学抗癌特性。为此，在一系列正常和癌细胞系中评估了它们在细胞和分子水平上的特性。通过进行细胞毒性测定和逆转录定量聚合酶链反应的基因表达分析获得的结果表明：i）在所分析的倍半萜衍生物中，VDS58 在附着（U-87 MG 和 MCF-7）和悬浮（K562 和 MEL-745）癌细胞培养物中表现出显著的抗癌特性；然而，U-87 MG 细胞在暴露 48 小时后能够迅速恢复其增殖能力；ii）与 K562 细胞相比，U-87 MG 细胞的基因表达谱显示细胞周期蛋白依赖性激酶抑制剂 1A（CDKN1）表达短暂诱导；iii）VDS58 处理 U-87 MG 细胞 12 小时后，转化生长因子β1（TGFB1）的表达水平升高，并在整个治疗期间保持在该水平；iv）在 K562 细胞中，VDS58 暴露 48 小时后 TGFB1 表达水平上调，随后下降；v）在先用 VDS58 预处理的 U-87 MG 培养物中再添加 VDS58，导致半胱天冬酶（CASP3 和 CASP9）、细胞死亡相关的 BCL2 激动剂（BAD）、细胞周期蛋白 D1、CDK6、CDKN1、原癌基因 bHLH 转录因子（MYC）、TGFB1 和肿瘤抑制蛋白 p53 的表达显著增加。大多数基因的这种上调仅持续 24 小时，此后仅 TGFB1 和 MYC 的表达保持高水平。通过对亲本 U-87 MG 和 K562 细胞的 RNA-Seq 数据进行生物信息学途径分析，报告了参与细胞周期调节的基因表达谱的显著变化。这与相同细胞暴露于 VDS58 时观察到的不同药理学特征相关。总体而言，本研究提供了有关倍半萜衍生物通过调节癌细胞周期相关基因的表达水平发挥作用的分子机制的新见解。
