Boueroy Parichart, Hahnvajanawong Chariya, Boonmars Thidarut, Saensa-ard Sunitta, Wattanawongdon Wareeporn, Kongsanthia Charuphan, Salao Kanin, Wongwajana Suwin, Anantachoke Natthinee, Reutrakul Vichai
Department of Microbiology, Khon Kaen University, Khon Kaen 40002, Thailand.
Liver Fluke and Cholangiocarcinoma Research Center, Cholangiocarcinoma Screening and Care Program (CASCAP), Khon Kaen 40002, Thailand.
Asian Pac J Cancer Prev. 2017 Dec 29;18(12):3343-3351. doi: 10.22034/APJCP.2017.18.12.3343.
Background: Chemotherapy for advanced cholangiocarcinoma (CCA) is largely ineffective; thus innovative combinations of chemotherapeutic agents and natural compounds represent a promising strategy. This study aimed to investigate the synergistic effects of forbesione combined with 5-fluorouracil (5-FU) in hamster cholangiocarcinoma (Ham-1) cells both in vitro and in vivo. The anti-tumor effects of 5-FU combined with forbesione in vitro were determined using the Sulforhodamine B (SRB) assay and the effects in vivo were assessed in transplanted Ham-1 allograph models. Using ethidium bromide/acridine orange (EB/AO) staining, the morphological changes of apoptotic cells was investigated. The expressions of apoptosis-related molecules after combined treatment with forbesione and 5-FU were determined using real-time RT-PCR and western blot analysis. Forbesione or 5-FU alone inhibited proliferation of Ham-1 cells in a dose-dependent manner and their combination showed a synergistic proliferation inhibitory effect in vitro. In vivo studies, forbesione in combination with 5-FU exhibited greater inhibition of the tumor in the hamster model compared with treatment using either drug alone. Forbesione combined with 5-FU exerted stronger apoptotic induction in Ham-1 cells than did single drug treatment. The combination of drugs strongly suppressed the expression of B-cell lymphoma 2 (Bcl-2) and procaspase-3 while enhancing the expression of p53, Bcl-2-associated X protein (Bax), apoptotic protease activating factor-1 (Apaf-1), caspase-9 and caspase-3, compared with single drug treatments. These results explained the decreased expression of cytokeratin 19 (CK19) positive cells and proliferation cell nuclear antigen (PCNA) positive cells in Ham-1 cell tumor tissues of the treated hamsters. There was no apparent systemic toxicity observed in the treated animals compared with the control groups. Forbesione combined with 5-FU strongly induced apoptosis in Ham-1 cells. The growth inhibitory effect of combined treatment using these two drugs was much greater than treatment with either drug alone, both in vitro and in vivo.
晚期胆管癌（CCA）的化疗大多无效；因此，化疗药物与天然化合物的创新组合是一种很有前景的策略。本研究旨在探讨福贝西酮与5-氟尿嘧啶（5-FU）联合应用对仓鼠胆管癌（Ham-1）细胞的体内外协同作用。采用磺酰罗丹明B（SRB）法测定5-FU与福贝西酮联合应用的体外抗肿瘤作用，并在移植的Ham-1同种异体模型中评估其体内作用。使用溴化乙锭/吖啶橙（EB/AO）染色，研究凋亡细胞的形态变化。采用实时逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析，测定福贝西酮与5-FU联合处理后凋亡相关分子的表达。单独使用福贝西酮或5-FU均能以剂量依赖的方式抑制Ham-1细胞的增殖，二者联合应用在体外表现出协同的增殖抑制作用。在体内研究中，与单独使用任一药物相比，福贝西酮与5-FU联合应用在仓鼠模型中对肿瘤的抑制作用更强。福贝西酮与5-FU联合应用比单一药物治疗更能强烈诱导Ham-1细胞凋亡。与单一药物治疗相比，联合用药强烈抑制B细胞淋巴瘤2（Bcl-2）和前半胱天冬酶-3的表达，同时增强p53、Bcl-2相关X蛋白（Bax）、凋亡蛋白酶激活因子-1（Apaf-1）、半胱天冬酶-9和半胱天冬酶-3的表达。这些结果解释了经处理仓鼠的Ham-1细胞肿瘤组织中细胞角蛋白19（CK19）阳性细胞和增殖细胞核抗原（PCNA）阳性细胞表达的降低。与对照组相比，在接受治疗的动物中未观察到明显的全身毒性。福贝西酮与5-FU联合应用能强烈诱导Ham-1细胞凋亡。这两种药物联合治疗的生长抑制作用在体外和体内均远大于单一药物治疗。
