Department of Pathology, Faculty of Medicine and Odontology, University of Valencia, Valencia, Spain.
Research Institute of the University Clinical Hospital of Valencia (INCLIVA), Valencia, Spain.
PLoS One. 2020 Feb 12;15(2):e0228909. doi: 10.1371/journal.pone.0228909. eCollection 2020.
BACKGROUND/OBJECTIVE: Patients with non-small cell lung cancer (NSCLC) develop resistance to antitumor agents by mechanisms that involve the epithelial-to-mesenchymal transition (EMT). This necessitates the development of new complementary drugs, e.g., cannabinoid receptors (CB1 and CB2) agonists including tetrahydrocannabinol (THC) and cannabidiol (CBD). The combined use of THC and CBD confers greater benefits, as CBD enhances the effects of THC and reduces its psychotropic activity. We assessed the relationship between the expression levels of CB1 and CB2 to the clinical features of a cohort of patients with NSCLC, and the effect of THC and CBD (individually and in combination) on proliferation, EMT and migration in vitro in A549, H460 and H1792 lung cancer cell lines.
Expression levels of CB1, CB2, EGFR, CDH1, CDH2 and VIM were evaluated by quantitative reverse transcription-polymerase chain reaction. THC and CBD (10-100 μM), individually or in combination (1:1 ratio), were used for in vitro assays. Cell proliferation was determined by BrdU incorporation assay. Morphological changes in the cells were visualized by phase-contrast and fluorescence microscopy. Migration was studied by scratch recolonization induced by 20 ng/ml epidermal growth factor (EGF).
The tumor samples were classified according to the level of expression of CB1, CB2, or both. Patients with high expression levels of CB1, CB2, and CB1/CB2 showed increased survival reaching significance for CB1 and CB1/CB2 (p = 0.035 and 0.025, respectively). Both cannabinoid agonists inhibited the proliferation and expression of EGFR in lung cancer cells, and CBD potentiated the effect of THC. THC and CBD alone or in combination restored the epithelial phenotype, as evidenced by increased expression of CDH1 and reduced expression of CDH2 and VIM, as well as by fluorescence analysis of cellular cytoskeleton. Finally, both cannabinoids reduced the in vitro migration of the three lung cancer cells lines used.
The expression levels of CB1 and CB2 have a potential use as markers of survival in patients with NSCLC. THC and CBD inhibited the proliferation and expression of EGFR in the lung cancer cells studied. Finally, the THC/CBD combination restored the epithelial phenotype in vitro.
背景/目的：非小细胞肺癌（NSCLC）患者通过涉及上皮-间质转化（EMT）的机制对抗肿瘤药物产生耐药性。这就需要开发新的互补药物，例如大麻素受体（CB1 和 CB2）激动剂，包括四氢大麻酚（THC）和大麻二酚（CBD）。THC 和 CBD 的联合使用带来了更大的益处，因为 CBD 增强了 THC 的作用并降低了其精神活性。我们评估了 CB1 和 CB2 的表达水平与 NSCLC 患者队列的临床特征之间的关系，以及 THC 和 CBD（单独使用和联合使用）对体外 A549、H460 和 H1792 肺癌细胞系增殖、EMT 和迁移的影响。
通过定量逆转录-聚合酶链反应评估 CB1、CB2、EGFR、CDH1、CDH2 和 VIM 的表达水平。单独或联合（1:1 比例）使用 10-100 μM 的 THC 和 CBD 进行体外测定。通过 BrdU 掺入测定法测定细胞增殖。通过 20ng/ml 表皮生长因子（EGF）诱导的划痕再殖民化研究迁移。
根据 CB1、CB2 或两者的表达水平对肿瘤样本进行分类。CB1、CB2 和 CB1/CB2 高表达水平的患者生存时间延长，CB1 和 CB1/CB2 均具有统计学意义（p=0.035 和 0.025）。两种大麻素激动剂均抑制肺癌细胞中 EGFR 的增殖和表达，CBD 增强了 THC 的作用。单独使用 THC 和 CBD 或联合使用可恢复上皮表型，表现为 CDH1 表达增加，CDH2 和 VIM 表达减少，以及细胞细胞骨架的荧光分析。最后，两种大麻素均减少了三种肺癌细胞系的体外迁移。
CB1 和 CB2 的表达水平有可能作为 NSCLC 患者生存的标志物。THC 和 CBD 抑制了研究中肺癌细胞中 EGFR 的增殖和表达。最后，THC/CBD 联合在体外恢复了上皮表型。
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