PDF(Pubmed)
Abstract：
Current treatment options available for prostate cancer (PCa) patients have many adverse side effects and hence, new alternative therapies need to be explored. Anticancer potential of various phytochemicals derived from Calotropis procera has been studied in many cancers but no study has investigated the effect of leaf extract of C. procera on PCa cells. Hence, we investigated the effect of C. procera leaf extract (CPE) on cellular properties of androgen-independent PC-3 and androgen-sensitive 22Rv1 cells. A hydroalcoholic extract of C. procera was prepared and MTT assay was performed to study the effect of CPE on viability of PCa cells. The effect of CPE on cell division ability, migration capability and reactive oxygen species (ROS) production was studied using colony formation assay, wound-healing assay and 2\',7\'-dichlorodihydrofluorescein diacetate assay, respectively. Caspase activity assay and LDH assay were performed to study the involvement of apoptosis and necrosis in CPE-mediated cell death. Protein levels of cell cycle, antioxidant, autophagy and apoptosis markers were measured by western blot. The composition of CPE was identified using untargeted LC-MS analysis. Results showed that CPE decreased the viability of both the PCa cells, PC-3 and 22Rv1, in a dose- and time-dependent manner. Also, CPE significantly inhibited the colony-forming ability, migration and endogenous ROS production in both the cell lines. Furthermore, CPE significantly decreased NF-κB protein levels and increased the protein levels of the cell cycle inhibitor p27. A significant increase in expression of autophagy markers was observed in CPE-treated PC-3 cells while autophagy markers were downregulated in 22Rv1 cells after CPE exposure. Hence, it can be concluded that CPE inhibits PCa cell viability possibly by regulating the autophagy pathway and/or altering the ROS levels. Thus, CPE can be explored as a possible alternative therapeutic agent for PCa.
摘要：
目前可用于前列腺癌（PCa）患者的治疗选择有许多不良副作用，因此，需要探索新的替代疗法。已在许多癌症中研究了源自Calotropisprocera的各种植物化学物质的抗癌潜力，但尚无研究研究C.procera的叶提取物对PCa细胞的影响。因此，我们研究了C.procera叶提取物（CPE）对雄激素非依赖性PC-3和雄激素敏感性22Rv1细胞的细胞特性的影响。制备C.procera的水醇提取物并进行MTT测定以研究CPE对PCa细胞活力的影响。CPE对细胞分裂能力的影响,使用集落形成试验研究了迁移能力和活性氧（ROS）的产生，伤口愈合试验和2'，7\'-二氯二氢荧光素二乙酸酯测定，分别。进行了Caspase活性测定和LDH测定，以研究凋亡和坏死在CPE介导的细胞死亡中的作用。细胞周期的蛋白质水平，抗氧化剂，通过westernblot检测自噬和凋亡标志物。使用非靶向LC-MS分析鉴定CPE的组成。结果表明，CPE降低了PCa细胞的活力，PC-3和22Rv1，以剂量和时间依赖的方式。此外，CPE显著抑制菌落形成能力,两种细胞系中的迁移和内源性ROS产生。此外，CPE显着降低NF-κB蛋白水平并增加细胞周期抑制剂p27的蛋白水平。在CPE处理的PC-3细胞中观察到自噬标志物的表达显著增加,而在CPE暴露后22Rv1细胞中自噬标志物下调。因此，可以得出结论，CPE可能通过调节自噬途径和/或改变ROS水平来抑制PCa细胞活力。因此，可以探索CPE作为PCa的可能的替代治疗剂。
