Department of Biosciences, Integral University, Dasauli, Kursi Road, Lucknow, 226026, India.
Noida Institute of Engineering and Technology, 19, Knowledge Park-II, Institutional Area, Greater Noida, 201306, India.
Anticancer Agents Med Chem. 2019;19(13):1588-1608. doi: 10.2174/1871520619666190731152942.
Several studies have revealed that abnormal activation of Notch signaling is closely related with the development and progression of prostate cancer. Although there are numerous therapeutic strategies, a more effective modality with least side effects is urgently required for the treatment of prostate cancer. Carvacrol is a monoterpenoid phenol and majorly present in the essential oils of Lamiaceae family plants. Many previous reports have shown various biological activities of carvacrol like antioxidant, antiinflammatory and anticancer properties. Recently, we have shown potent anticancer property of carvacrol against prostate cancer cell line DU145. In the current study, we report the chemopreventive and therapeutic potential of carvacrol against another prostate cancer cell line PC-3 with its detailed mechanism of action.
To determine the effect of the carvacrol on prostate cancer cells, the cell viability was estimated by MTT assay and cell death was estimated by LDH release assay. The apoptotic assay was performed by DAPI staining and FITC-Annexin V assay. Reactive Oxygen Species (ROS) was estimated by DCFDA method. Cell cycle analysis was performed by flow cytometry. Gene expression analysis was performed by quantitative real time PCR.
Our results suggested that the carvacrol treatment significantly reduced the cell viability of PC-3 cells in a dose- and time-dependent manner. The antiproliferative action of carvacrol was correlated with apoptosis which was confirmed by nuclear condensation, FITC-Annexin V assay, modulation in expression of Bax, Bcl-2 and caspase activation. The mechanistic insight into carvacrol-induced apoptosis leads to finding of elevated level of Reactive Oxygen Species (ROS) and mitochondrial membrane potential disruption. Cell cycle analysis revealed that carvacrol prevented cell cycle in G0/G1 that was associated with decline in expression of cyclin D1 and Cyclin-Dependent Kinase 4 (CDK4) and augmented expression of CDK inhibitor p21. Having been said the role of hyperactivation of Notch signaling in prostate cancer, we also deciphered that carvacrol could inhibit Notch signaling in PC-3 cells via downregulation of Notch-1, and Jagged-1.
Thus, our previous and current findings have established the strong potential of carvacrol as a chemopreventive agent against androgen-independent human prostate cancer cells.
多项研究表明，Notch 信号的异常激活与前列腺癌的发生和发展密切相关。尽管有许多治疗策略，但对于前列腺癌的治疗，仍迫切需要更有效且副作用更小的方法。香芹酚是单萜酚类化合物，主要存在于唇形科植物的精油中。许多先前的报告显示，香芹酚具有多种生物学活性，如抗氧化、抗炎和抗癌特性。最近，我们已经证明香芹酚对前列腺癌细胞系 DU145 具有强大的抗癌作用。在本研究中，我们报告了香芹酚对另一种前列腺癌细胞系 PC-3 的化学预防和治疗潜力，并详细阐述了其作用机制。
为了确定香芹酚对前列腺癌细胞的影响，通过 MTT 测定法评估细胞活力，通过 LDH 释放测定法评估细胞死亡。通过 DAPI 染色和 FITC-Annexin V 测定法进行凋亡测定。通过 DCFDA 法评估活性氧 (ROS)。通过流式细胞术进行细胞周期分析。通过定量实时 PCR 进行基因表达分析。
我们的结果表明，香芹酚处理以剂量和时间依赖的方式显著降低了 PC-3 细胞的细胞活力。香芹酚的抗增殖作用与凋亡相关，这通过核浓缩、FITC-Annexin V 测定、Bax、Bcl-2 表达的调节和半胱天冬酶激活得到证实。香芹酚诱导凋亡的机制研究导致发现活性氧 (ROS) 水平升高和线粒体膜电位破坏。细胞周期分析显示，香芹酚阻止细胞周期进入 G0/G1 期，这与细胞周期蛋白 D1 和细胞周期蛋白依赖性激酶 4 (CDK4) 的表达下降以及 CDK 抑制剂 p21 的表达增加有关。由于 Notch 信号的过度激活在前列腺癌中的作用，我们还发现香芹酚可以通过下调 Notch-1 和 Jagged-1 来抑制 PC-3 细胞中的 Notch 信号。
因此，我们之前和现在的研究结果已经确立了香芹酚作为雄激素非依赖性人前列腺癌细胞化学预防剂的强大潜力。
