Melatonin potentiates cisplatin-induced apoptosis and cell cycle arrest in human lung adenocarcinoma cells
Melatonin potentiates cisplatin-induced apoptosis and cell cycle arrest in human lung adenocarcinoma cells
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DOI：
10.1111/cpr.12158
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发表时间：
2015-02-01
影响因子：
8.5
通讯作者：
Johns, N. P.
中科院分区：
文献类型：
--
作者：
Plaimee, P.;Weerapreeyakul, N.;Johns, N. P.
目的褪黑素通过多种机制发挥抗癌作用，包括诱导细胞凋亡。通过这种方式，它已被证明与化疗药物一起用于癌症治疗。本文研究了褪黑素对化疗药物顺铂诱导的人肺腺癌顺铂敏感细胞株(SK-LU-1)的细胞毒作用、细胞周期停滞和细胞凋亡的影响。材料与方法：用1-5 mM的褪黑素或10-200M的顺铂单独作用于SK-LU-1细胞48h。用四甲基偶氮唑蓝还原比色法测定细胞毒作用。流式细胞仪Annexin V/PI染色和DAPI核染色检测细胞凋亡。用DiOC6(3)试剂测定线粒体膜电位m的变化，并检测caspase-3/7的活性。结果5 mM和34M浓度的褪黑素和顺铂单独作用时，细胞毒作用IC50为50%。1 mM或2 mM褪黑素与顺铂联合应用可显著增强顺铂的细胞毒作用，使其IC50分别降至11和4M。与单用顺铂相比，联合用药通过提高线粒体膜去极化、激活caspase-3/7和诱导细胞周期停滞于S时相一致地增加了凋亡细胞的数量。结论褪黑素增强了顺铂诱导的SK-LU-1肺癌细胞的细胞毒作用和细胞凋亡。SK-LU-1细胞群体生长抑制是通过细胞周期停滞于S期而实现的。这些发现表明，褪黑素有可能与顺铂等化疗药物联合用于非小细胞肺癌的治疗。
ObjectivesMelatonin produces anti-cancer effects via several mechanisms, including by induction of apoptosis. In this way, it has been shown to be of use, in combination with chemotherapeutic drugs, for cancer treatment. The study described here has evaluated effects of melatonin on cytotoxicity, apoptosis and cell cycle arrest induced with the chemotherapeutic agent cisplatin, in human lung adenocarcinoma cisplatin-sensitive cell line (SK-LU-1), which previously had only limit data.Materials and methodsCells of the SK-LU-1 line were treated with melatonin alone at 1-5mm concentration or cisplatin alone 10-200m, for 48h in culture. Cytotoxicity was measured by MTT reduction assay. Apoptosis induction was detected by annexin V/PI staining using flow cytometric analysis and DAPI nuclear staining. Change in mitochondrial membrane potential (m) was quantified using DiOC6(3) reagent and activities of caspases-3/7 were also investigated. DNA fractions were measured using propidium iodide (PI) staining.ResultsMelatonin or cisplatin alone had 50% (IC50) cytotoxicity at 5mm or 34m concentrations respectively. Combination of 1 or 2mm melatonin and cisplatin significantly augmented cytotoxicity of cisplatin by reducing its IC50 to 11 and 4m, respectively. Consistently, combined treatment increased population of apoptotic cells by elevating mitochondrial membrane depolarization, activating caspases-3/7 and inducing cell cycle arrest in the S phase, compared to treatment with cisplatin alone.ConclusionThese data demonstrate that melatonin enhanced cisplatin-induced cytotoxicity and apoptosis in SK-LU-1 lung cancer cells. SK-LU-1 cell population growth inhibition was mediated by cell cycle arrest in the S phase. These findings suggest that melatonin has the potential to be used for NSCLC treatment in combination with a chemotherapeutic agent such as cisplatin.