Ahlina Faradiba Nur, Nugraheni Nadzifa, Salsabila Irfani Aura, Haryanti Sari, Da'i Muhammad, Meiyanto Edy
Cancer Chemoprevention Research Center, Faculty of Pharmacy, Universitas Gadjah Mada, Indonesia.
Medicinal Plant and Traditional Medicine Research and Development Centre, Ministry of Health, Indonesia.
Asian Pac J Cancer Prev. 2020 Jan 1;21(1):107-117. doi: 10.31557/APJCP.2020.21.1.107.
This study intends to explore the potential of galangal extract as a co-chemotherapeutic agent through the analysis of its cytotoxic and migratory effects on metastatic breast cancer cells and as an anti-ageing agent through its senescence inhibitory effect on normal fibroblast cells.
Galangal ethanolic extract (GE) was subjected to a cytotoxicity test with the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay alone or in combination with doxorubicin (Dox) against 4T1 cells but not in NIH-3T3 cells. Evidence of senescent cells was detected using a SA-β galactosidase based assay. In addition, the level of reactive oxygen species (ROS), apoptosis, and cell cycle were measured with a flow cytometry-based assay. Meanwhile, cell migration and matrix metalloproteinase (MMP)-9 expression after GE treatment on 4T1 cells were measured using the scratch wound healing assay and gelatin zymography assay, respectively. The metabolomic profiles of GE were traced using gas chromatography-mass spectrometry (GC-MS) analysis.
GE effectively inhibited the growth of 4T1 cells with an IC50 value of 135 µg/mL and increased the cytotoxic effect of Dox at concentrations of 50 and 100 µg/mL. GE increased the number of senescent cells arrested in the G2/M phase but did not cause apoptosis. This effect is compounded by increasing intracellular levels of ROS. However, GE reduced senescence to normal in fibroblast cells (NIH 3T3 cells) under oxidative stress by Dox without any changes in the ROS level. Moreover, GE also inhibited the migration of 4T1 cells and suppressed the expression of MMP-9 induced by Dox.
Galangal has the potential for use as a co-chemotherapeutic agent by inducing senescence in correlation with increasing intracellular ROS toward metastatic breast cancer. However, the effect of GE in decreasing the senescence phenomena toward normal fibroblast cells illustrates its potential as a promising anti-ageing agent.
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本研究旨在通过分析高良姜提取物对转移性乳腺癌细胞的细胞毒性和迁移作用，探索其作为辅助化疗药物的潜力，并通过其对正常成纤维细胞的衰老抑制作用，探索其作为抗衰老药物的潜力。
采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法单独或与阿霉素（Dox）联合检测高良姜乙醇提取物（GE）对4T1细胞的细胞毒性，但不检测对NIH-3T3细胞的毒性。使用基于SA-β半乳糖苷酶的检测方法检测衰老细胞的证据。此外，通过基于流式细胞术的检测方法测量活性氧（ROS）水平、细胞凋亡和细胞周期。同时，分别使用划痕伤口愈合试验和明胶酶谱分析检测GE处理后4T1细胞的迁移和基质金属蛋白酶（MMP）-9表达。使用气相色谱-质谱（GC-MS）分析追踪GE的代谢组学特征。
GE有效抑制4T1细胞生长，IC50值为135μg/mL，并在50和100μg/mL浓度下增强了Dox的细胞毒性作用。GE增加了停滞在G2/M期的衰老细胞数量，但未引起细胞凋亡。这种作用因细胞内ROS水平升高而加剧。然而，GE在Dox诱导的氧化应激下使成纤维细胞（NIH 3T3细胞）中的衰老恢复正常，而ROS水平没有任何变化。此外，GE还抑制了4T1细胞的迁移，并抑制了Dox诱导的MMP-9表达。
高良姜有潜力通过诱导衰老并增加细胞内ROS来作为辅助化疗药物用于转移性乳腺癌。然而，GE对正常成纤维细胞衰老现象的减轻作用表明其作为一种有前景的抗衰老药物的潜力。
