Department of Urology, Affiliated Dongguan People's Hospital, Southern Medical University, Dongguan 523059, Guangdong, P.R. China.
J BUON. 2020 Jan-Feb;25(1):280-285.
The primary purpose of the current study was to investigate the antitumor activity of limonene which is a plant monoterpene along with evaluating its effects on cell apoptosis, cell cycle phase distribution, cell migration and invasion.
The cell proliferation of T24 bladder cancer cells was examined by WTS-1 assay. The apoptotic effects induced by limonene were investigated by a combination of fluorescence microscopy and flow cytometry and then confirmed by western blot assay. The effects of limonene on cell cycle in T24 bladder cancer cells were studied by flow cytometry. The effects on cell migration and invasion were examined by wound healing assay and transwell assay using Matrigel.
The results showed that limonene induced cytotoxic effects and reduced cell viability of T24 human bladder cancer cells showing an IC50 value of 9 μM. Limonene also induced significant apoptosis in bladder cancer cells since it induced significant nuclear fragmentation, chromatin condensation, and splitting of the nucleus, representative of the apoptotic cascade. The apoptotic cell percentage was 1.95, 5.35, 15.61 and 34.71% at limonene concentrations of 0, 9, 18 and 36 µM. Further, the apoptotic effects of limonene were also confirmed by Western blot analysis and the results showed increase in the expression of Bax and caspase-3 and decrease of Bcl-2 expression. Limonene also caused G2/M phase cell cycle arrest as well as suppression of cell migration and invasion.
These results indicate that limonene might be used as a potent anticancer agent against human bladder cancer for which further in depth studies are needed, especially over its toxicological studies.
本研究的主要目的是研究柠檬烯作为植物单萜烯的抗肿瘤活性，同时评估其对细胞凋亡、细胞周期分布、细胞迁移和侵袭的影响。
通过 WTS-1 测定法检测 T24 膀胱癌细胞的细胞增殖。通过荧光显微镜和流式细胞术相结合，研究柠檬烯诱导的细胞凋亡作用，并通过 Western blot 检测进一步证实。通过流式细胞术研究柠檬烯对 T24 膀胱癌细胞周期的影响。通过划痕愈合试验和 Transwell 试验（使用 Matrigel）研究柠檬烯对细胞迁移和侵袭的影响。
结果表明，柠檬烯诱导 T24 人膀胱癌细胞的细胞毒性作用，降低细胞活力，IC50 值为 9 μM。柠檬烯还诱导膀胱癌细胞发生显著的细胞凋亡，因为它诱导了明显的核碎裂、染色质浓缩和核分裂，代表了凋亡级联反应。在 0、9、18 和 36 μM 的柠檬烯浓度下，凋亡细胞的百分比分别为 1.95%、5.35%、15.61%和 34.71%。此外，Western blot 分析也证实了柠檬烯的凋亡作用，结果显示 Bax 和 caspase-3 的表达增加，Bcl-2 的表达减少。柠檬烯还导致 G2/M 期细胞周期停滞以及细胞迁移和侵袭的抑制。
这些结果表明，柠檬烯可能作为一种有效的抗癌剂用于人类膀胱癌，需要进一步深入研究，特别是在其毒理学研究方面。
