■前列腺癌(PCa)是全世界男性死亡的原因之一。尽管已经制定了治疗策略,该疾病的复发和随之而来的副作用仍然是一个重要的问题。杜松子酒,一种传统的泰国药物,表现出不同的治疗特性,包括抗癌活性。然而,其对前列腺癌的抗癌活性尚未得到彻底探索。本研究旨在评估与LNCaP人前列腺癌细胞系中凋亡诱导相关的杜鹃乙酸乙酯提取物(EADR)的抗癌活性和潜在机制。
■乙酸乙酯用于提取杜鹃花的干燥树皮。使用MTS测定评估EADR对LNCaP和WPMY-1细胞(正常人前列腺成肌纤维细胞系)的细胞毒性。EADR对细胞周期的影响,凋亡诱导,通过碘化丙啶(PI)染色评估线粒体膜电位(MMP)的变化,膜联蛋白V-FITC/PI,和JC-1染料,分别。随后使用流式细胞术进行分析。裂解的caspase-3,BAX,和Bcl-2通过蛋白质印迹检查。EADR的植物化学分析使用气相色谱-质谱(GC-MS)进行。
■EADR对LNCaP细胞表现出剂量依赖性的细胞毒作用,24小时和48小时后的IC50值为15.43和12.35µg/mL,分别。尽管它对WPMY-1细胞也表现出细胞毒性作用,影响相对较低,暴露24和48小时后的IC50值分别为34.61和19.93µg/mL,分别。细胞周期分析表明,EADR在LNCaP或WPMY-1细胞中均未诱导细胞周期停滞。然而,它显著增加了LNCaP细胞中的亚G1群体,表明细胞凋亡的潜在诱导。膜联蛋白V-FITC/PI染色表明EADR显著诱导LNCaP细胞凋亡。随后对EADR诱导的细胞凋亡的潜在机制的研究表明,JC-1染色证明了MMP的减少。此外,Westernblotting显示EADR治疗导致BAX上调,LNCaP细胞中BCL-2的下调和caspase-3裂解的升高。值得注意的是,通过GC-MS鉴定,在EADR中,epilupeol是一种突出的化合物。
■EADR通过诱导细胞毒性和细胞凋亡表现出对LNCaP人前列腺癌细胞系的抗癌活性。我们的发现表明EADR通过上调促凋亡BAX促进细胞凋亡,而抗凋亡Bcl-2的下调导致MMP的减少和caspase-3的激活。特别令人感兴趣的是epilupeol的存在,EADR中确定的主要化合物,这可能有望成为前列腺癌治疗药物开发的候选药物。
UNASSIGNED: Prostate cancer (PCa) is one of the causes of death in men worldwide. Although treatment strategies have been developed, the recurrence of the disease and consequential side effects remain an essential concern. Diospyros rhodocalyx Kurz, a traditional Thai medicine, exhibits diverse therapeutic properties, including anti-cancer activity. However, its anti-cancer activity against prostate cancer has not been thoroughly explored. This study aims to evaluate the anti-cancer activity and underlying mechanisms of the ethyl acetate extract of D. rhodocalyx Kurz (EADR) related to apoptosis induction in the LNCaP human prostate cancer cell line.
UNASSIGNED: Ethyl acetate was employed to extract the dried bark of D. rhodocalyx Kurz. The cytotoxicity of EADR on both LNCaP and WPMY-1 cells (normal human prostatic myofibroblast cell line) was evaluated using MTS assay. The effect of EADR on the cell cycle, apoptosis induction, and alteration in mitochondrial membrane potential (MMP) was assessed by the staining with propidium iodide (PI), Annexin V-FITC/PI, and JC-1 dye, respectively. Subsequent analysis was conducted using flow cytometry. The expression of cleaved caspase-3, BAX, and Bcl-2 was examined by Western blotting. The phytochemical profiling of the EADR was performed using gas chromatography-mass spectrometry (GC-MS).
UNASSIGNED: EADR exhibited a dose-dependent manner cytotoxic effect on LNCaP cells, with IC50 values of 15.43 and 12.35 µg/mL after 24 and 48 h, respectively. Although it also exhibited a cytotoxic effect on WPMY-1 cells, the effect was comparatively lower, with the IC50 values of 34.61 and 19.93 µg/mL after 24 and 48 h of exposure, respectively. Cell cycle analysis demonstrated that EADR did not induce cell cycle arrest in either LNCaP or WPMY-1 cells. However, it significantly increased the sub-G1 population in LNCaP cells, indicating a potential induction of apoptosis. The Annexin V-FITC/PI staining indicated that EADR significantly induced apoptosis in LNCaP cells. Subsequent investigation into the underlying mechanism of EADR-induced apoptosis revealed a reduction in MMP as evidenced by JC-1 staining. Moreover, Western blotting demonstrated that EADR treatment resulted in the upregulation of BAX, downregulation of BCL-2, and elevation of caspase-3 cleavage in LNCaP cells. Notably, the epilupeol was a prominent compound in EADR as identified by GC-MS.
UNASSIGNED: The EADR exhibits anti-cancer activity against the LNCaP human prostate cancer cell line by inducing cytotoxicity and apoptosis. Our findings suggest that EADR promotes apoptosis by upregulating pro-apoptotic BAX, whereas downregulation of anti-apoptotic Bcl-2 results in the reduction of MMP and the activation of caspase-3. Of particular interest is the presence of epilupeol, a major compound identified in EADR, which may hold promise as a candidate for the development of therapeutic agents for prostate cancer.