Department of General Surgery, Taizhou First People's Hospital in Zhejiang Province, Taizhou, Zhejiang 318020, P.R. China.
Department of Microbiology, School of Biological Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia.
Mol Med Rep. 2019 Jun;19(6):5368-5376. doi: 10.3892/mmr.2019.10201. Epub 2019 Apr 30.
Cancer chemotherapy possesses high toxicity, particularly when a higher concentration of drugs is administered to patients. Therefore, searching for more effective compounds to reduce the toxicity of treatments, while still producing similar effects as current chemotherapy regimens, is required. Currently, the search for potential anticancer agents involves a random, inaccurate process with strategic deficits and a lack of specific targets. For this reason, the initial in vitro high‑throughput steps in the screening process should be reviewed for rapid identification of the compounds that may serve as anticancer agents. The present study aimed to investigate the potential use of the Pichia pastoris strain SMD1168H expressing DNA topoisomerase I (SMD1168H‑TOPOI) in a yeast‑based assay for screening potential anticancer agents. The cell density that indicated the growth of the recombinant yeast without treatment was first measured by spectrophotometry. Subsequently, the effects of glutamate (agonist) and camptothecin (antagonist) on the recombinant yeast cell density were investigated using the same approach, and finally, the effect of camptothecin on various cell lines was determined and compared with its effect on recombinant yeast. The current study demonstrated that growth was enhanced in SMD1168H‑TOPOI as compared with that in SMD1168H. Glutamate also enhanced the growth of the SMD1168H; however, the growth effect was not enhanced in SMD1168H‑TOPOI treated with glutamate. By contrast, camptothecin caused only lower cell density and growth throughout the treatment of SMD1168H‑TOPOI. The findings of the current study indicated that SMD1168H‑TOPOI has similar characteristics to MDA‑MB‑231 cells; therefore, it can be used in a yeast‑based assay to screen for more effective compounds that may inhibit the growth of highly metastatic breast cancer cells.
癌症化疗具有很高的毒性,尤其是当给患者施用更高浓度的药物时。因此,需要寻找更有效的化合物来降低治疗毒性,同时仍能产生与当前化疗方案相似的效果。目前,寻找潜在的抗癌药物涉及一个随机的、不准确的过程,存在战略缺陷和缺乏特定的靶点。因此,应该对筛选过程中的初始体外高通量步骤进行审查,以便快速识别可能作为抗癌剂的化合物。本研究旨在探讨表达 DNA 拓扑异构酶 I 的毕赤酵母菌株 SMD1168H(SMD1168H-TOPOI)在酵母基础测定中筛选潜在抗癌药物的潜在用途。首先通过分光光度法测量指示重组酵母无处理生长的细胞密度。随后,采用相同方法研究谷氨酸(激动剂)和喜树碱(拮抗剂)对重组酵母细胞密度的影响,最后,确定喜树碱对各种细胞系的作用并将其与对重组酵母的作用进行比较。本研究表明,与 SMD1168H 相比,SMD1168H-TOPOI 的生长增强。谷氨酸也增强了 SMD1168H 的生长;然而,谷氨酸处理的 SMD1168H-TOPOI 的生长效果没有增强。相比之下,喜树碱仅导致 SMD1168H-TOPOI 整个治疗过程中的细胞密度和生长降低。本研究的结果表明,SMD1168H-TOPOI 具有与 MDA-MB-231 细胞相似的特征;因此,它可以用于酵母基础测定来筛选更有效的化合物,这些化合物可能抑制高转移性乳腺癌细胞的生长。