Department of Biotechnology, Medical University of Bialystok, Bialystok, Poland.
Department of Synthesis and Technology of Drugs, Medical University of Bialystok, Bialystok, Poland.
PLoS One. 2021 Aug 26;16(8):e0255585. doi: 10.1371/journal.pone.0255585. eCollection 2021.
Gastric cancer (GC) is a multifactorial disease with high mortality. Anti-HER2 therapy is a promising strategy in GC treatment and trastuzumab was approved by FDA (Food and Drug Administration) as the first and the second line of treatment of the disease.
The aim of the study was to examine the effectiveness of a combination of etoposide with trastuzumab or pertuzumab in AGS gastric cancer cells and breast cancer cells such as MCF-7, MDA-MB-231 and HCC1954.
The cytotoxic effects of the tested compounds against gastric and breast cancer cells were checked by MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assay. The anti-proliferative potential was analyzed by the incorporation of [3H]-thymidine into DNA. Fluorescent microscopy and flow cytometry was used to demonstrate the effect of the compounds on apoptosis. The mitochondrial membrane potential, and the activity of caspase-8 and caspase-9 were assessed. Autophagosomes and autolysosomes formation was checked by flow cytometry. The concentrations of Beclin-1, LC3A and LC3B were performed using ELISA. The expression of LC3A/B was also determined. The results from our study proved that the combination of etoposide with anti-HER2 antibodies was not cytotoxic against breast cancer cells, whereas the combination of etoposide with anti-HER2 antibodies decreased viability and DNA biosynthesis in gastric cancer cells. The interaction of etoposide with pertuzumab or trastuzumab induced programmed cell death via extrinsic and intrinsic apoptotic pathways in AGS gastric cancer cells, but did not affect autophagy, where a decrease of Beclin-1, LC3A and LC3B was observed in comparison with the untreated control.
The study demonstrated that etoposide (12.5 μM) with pertuzumab represent a promising strategy in gastric cancer treatment, but further in vivo examinations are also required.
胃癌（GC）是一种具有高死亡率的多因素疾病。抗 HER2 治疗是 GC 治疗的一种有前途的策略，曲妥珠单抗已被 FDA（美国食品和药物管理局）批准为该疾病的一线和二线治疗药物。
本研究旨在研究依托泊苷与曲妥珠单抗或帕妥珠单抗联合应用于 AGS 胃癌细胞和乳腺癌细胞（如 MCF-7、MDA-MB-231 和 HCC1954）的疗效。
通过 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）测定法检查测试化合物对胃癌和乳腺癌细胞的细胞毒性作用。通过将[3H]-胸苷掺入 DNA 中来分析抗增殖潜力。荧光显微镜和流式细胞术用于证明化合物对细胞凋亡的影响。评估线粒体膜电位以及 caspase-8 和 caspase-9 的活性。通过流式细胞术检查自噬体和自溶酶体的形成。使用 ELISA 检测 Beclin-1、LC3A 和 LC3B 的浓度。还确定了 LC3A/B 的表达。我们的研究结果证明，依托泊苷与抗 HER2 抗体的联合应用对乳腺癌细胞没有细胞毒性，而依托泊苷与抗 HER2 抗体的联合应用降低了胃癌细胞的活力和 DNA 生物合成。依托泊苷与 pertuzumab 或 trastuzumab 的相互作用通过外在和内在凋亡途径诱导 AGS 胃癌细胞程序性细胞死亡，但不影响自噬，与未处理的对照组相比，观察到 Beclin-1、LC3A 和 LC3B 的减少。
该研究表明，依托泊苷（12.5 μM）与 pertuzumab 联合应用是治疗胃癌的一种有前途的策略，但还需要进一步的体内研究。
