Department of Cell Biology, College of Basic Medicine, Capital Medical University, Beijing 100069, P.R. China.
Clinical Medical Research Center, The Affiliated Hospital of Inner Mongolia Medical University, Inner Mongolia Autonomous Region 010050, P.R. China.
Oncol Rep. 2020 Jan;43(1):201-217. doi: 10.3892/or.2019.7406. Epub 2019 Nov 11.
The combination of chemotherapeutic modalities may be more effective in treating gastric cancer compared with any modality alone. Previous studies have demonstrated that the combination of anticancer bioactive peptides (ACBP) and oxaliplatin (OXA) significantly inhibited the growth of the gastric cancer cell line MKN‑45, promoted the apoptosis of MKN‑45 cells, and caused an irreversible arrest of the MKN‑45 cell cycle in the G2/M phase. In the present study, an isobaric tag for relative and absolute quantitation (iTRAQ)‑based quantitative proteomics technique was used to determine the effect of ACBP‑OXA treatment on the proteomics profile of MKN‑45 cells. Notably, a total of 6,210 proteins were detected. Proteins with a >1.2‑fold change in expression (either up‑ or downregulation) and P<0.05 were considered to be differentially expressed. A total of 256 differentially expressed proteins were identified through alignments with different groups. Compared with the control group, MKN‑45 cells treated with ACBP, OXA and ACBP‑OXA exhibited 17 (10 up‑ and 7 downregulated), 111 (27 up‑ and 84 downregulated) and 128 (53 up‑ and 75 downregulated) differentially expressed proteins, respectively. Of the 256 differentially expressed proteins, 6 (TPX2, NUSAP1, TOP2A, YAP, MKi‑67 and GPC4) were verified by the parallel reaction monitoring method, which revealed that TPX2, NUSAP1, TOP2A, YAP, MKi‑67 and GPC4 expression decreased with ACBP‑OXA treatment. The cellular localization, functional annotation and biological pathways of differentially expressed proteins were examined by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis. The results indicated that ACBP‑OXA may act through the ribosome or the AMP‑activated protein kinase (AMPK) signaling pathway, and the AMPK signaling pathway may be an important mediator of the inhibitory effects of ACBP‑OXA on MKN‑45 gastric cancer cells. In summary, iTRAQ‑based proteomics analysis of the effect of ACBP‑OXA on MKN‑45 cells may guide future therapeutic strategies for gastric cancer. In addition, the present study may help provide new insights into the therapeutic role of combined ACBP and OXA in gastric cancer.
化疗方式的联合应用可能比任何单一方式更能有效治疗胃癌。先前的研究表明，抗癌生物活性肽（ACBP）与奥沙利铂（OXA）的联合应用显著抑制了胃癌细胞系 MKN-45 的生长，促进了 MKN-45 细胞的凋亡，并导致 MKN-45 细胞周期不可逆地阻滞在 G2/M 期。在本研究中，采用基于同位素相对和绝对定量标记（iTRAQ）的定量蛋白质组学技术来确定 ACBP-OXA 处理对 MKN-45 细胞蛋白质组图谱的影响。值得注意的是，共检测到 6210 种蛋白质。表达水平变化超过 1.2 倍（上调或下调）且 P<0.05 的蛋白被认为是差异表达蛋白。通过与不同组别的比对，共鉴定到 256 个差异表达蛋白。与对照组相比，ACBP、OXA 和 ACBP-OXA 处理的 MKN-45 细胞分别表现出 17 个（10 个上调和 7 个下调）、111 个（27 个上调和 84 个下调）和 128 个（53 个上调和 75 个下调）差异表达蛋白。在 256 个差异表达蛋白中，通过平行反应监测法验证了 6 个蛋白（TPX2、NUSAP1、TOP2A、YAP、MKi-67 和 GPC4）的表达下调。结果表明，随着 ACBP-OXA 的处理，TPX2、NUSAP1、TOP2A、YAP、MKi-67 和 GPC4 的表达下降。通过基因本体论和京都基因与基因组百科全书分析，对差异表达蛋白的细胞定位、功能注释和生物学途径进行了研究。结果表明，ACBP-OXA 可能通过核糖体或 AMP 激活蛋白激酶（AMPK）信号通路发挥作用，而 AMPK 信号通路可能是 ACBP-OXA 抑制 MKN-45 胃癌细胞的重要介导因子。综上所述，基于 iTRAQ 的 ACBP-OXA 对 MKN-45 细胞作用的蛋白质组学分析可能为胃癌的未来治疗策略提供指导。此外，本研究可能有助于为联合应用 ACBP 和 OXA 治疗胃癌提供新的见解。
