Harada Koji, Ferdous Tarannum, Kawasaki Keisuke, Watanabe Kenji, Mizukami Yoichi
Department of Nursing, Faculty of Health Sciences, Hiroshima Cosmopolitan University, Hiroshima 734-0014, Japan.
Department of Oral and Maxillofacial Surgery, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan.
Biomed Rep. 2025 Jul 1;23(3):150. doi: 10.3892/br.2025.2028. eCollection 2025 Sep.
Aged garlic extract (AGE) is one of the widely used garlic-based products that are extensively studied and are commercially available. However, there are no reports that compare the effects of AGE on normal cells and oral cancer cells. The present study tried to investigate the effects of AGE on normal healthy cells and cancer cells. The effect of AGE on the proliferation of normal cell lines HaCaT and fibroblast, in addition to cancer cell lines SCC7, HSC2, HSC3 and Ca9-22 were evaluated by MTT assay. The effects of AGE on cell migration were examined using wound healing and migration assays. Whole transcriptome analysis, ingenuity pathways analysis (IPA) and western blot analysis were used to investigate the mechanism of action of AGE in HaCaT and SCC7 cells. Data from the aforementioned assays were then evaluated and compared to assess if AGE affects normal cells differently compared with cancer cells. AGE was found to promote the proliferation and migration of normal cells, especially HaCaT, in the absence of FBS more markedly compared with those of cancer cells. However, AGE could not promote wound healing in fibroblast cells at the same rate as in HaCaT cells. In normal cells, sequential or combination AGE treatment with 5-fluorouracil (5-FU), cisplatin (CDDP) and docetaxel (DOC) somewhat counteracted the proliferation-limiting effects of anti-cancer agents. However, in cancer cells, AGE treatments enhanced the inhibitory effects of anti-cancer agents when used in combination with 5-FU, CDDP or DOC. This observation was more evident in the case of pre-treatment with anticancer agents followed by AGE sequential treatment. Subsequently, whole transcriptome analysis and IPA data suggested that AGE facilitated the proliferation and survival of normal cells through the induction of protein kinase B family protein and brain-derived neurotrophic factor pathways, whilst suppressing ferroptosis These data were also confirmed by western blotting to see if the genetic changes shown in whole transcriptome analysis and IPA are also induced at the protein level. In addition, AGE may reduce cancer cell proliferation through the suppression of cancer metastasis signaling and enhancement of phagocytic activity according to whole transcriptome analysis and IPA data. Taken together, these findings suggested that AGE may prompt the proliferation of normal cells and suppress the that of cancer cells through different cellular processes and signaling pathways.
老化大蒜提取物（AGE）是广泛使用的基于大蒜的产品之一，已被广泛研究且有商业销售。然而，尚无关于AGE对正常细胞和口腔癌细胞作用比较的报道。本研究试图探究AGE对正常健康细胞和癌细胞的影响。通过MTT法评估了AGE对正常细胞系HaCaT和成纤维细胞以及癌细胞系SCC7、HSC2、HSC3和Ca9-22增殖的影响。使用伤口愈合和迁移实验检测了AGE对细胞迁移的影响。采用全转录组分析、 Ingenuity通路分析（IPA）和蛋白质印迹分析来研究AGE在HaCaT和SCC7细胞中的作用机制。然后对上述实验数据进行评估和比较，以评估AGE对正常细胞和癌细胞的影响是否不同。结果发现，在无胎牛血清（FBS）的情况下，AGE促进正常细胞尤其是HaCaT细胞的增殖和迁移，其作用比癌细胞更为显著。然而，AGE促进成纤维细胞伤口愈合的速率不及HaCaT细胞。在正常细胞中，AGE与5-氟尿嘧啶（5-FU）、顺铂（CDDP）和多西他赛（DOC）序贯或联合处理在一定程度上抵消了抗癌药物的增殖限制作用。然而，在癌细胞中，AGE与5-FU, CDDP或DOC联合使用时增强了抗癌药物的抑制作用。这种现象在抗癌药物预处理后再进行AGE序贯处理的情况下更为明显。随后，全转录组分析和IPA数据表明，AGE通过诱导蛋白激酶B家族蛋白和脑源性神经营养因子通路促进正常细胞的增殖和存活，同时抑制铁死亡。蛋白质印迹也证实了这些数据，以查看全转录组分析和IPA中显示的基因变化是否也在蛋白质水平上被诱导。此外，根据全转录组分析和IPA数据，AGE可能通过抑制癌症转移信号和增强吞噬活性来减少癌细胞增殖。综上所述，这些发现表明，AGE可能通过不同的细胞过程和信号通路促进正常细胞增殖并抑制癌细胞增殖。
