Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University, Xining, Qinghai 810000, P.R. China.
Mol Med Rep. 2018 Mar;17(3):3735-3743. doi: 10.3892/mmr.2017.8346. Epub 2017 Dec 22.
Aspirin as an antitumor drug has been studied in various malignancies with regards to its effects on apoptosis, proliferation, metastasis and senescence of tumor cells. However, the clinical application is limited by its side effects. Nutlin‑3 is a novel antitumor compound, which has not been clinically approved. The present study investigated the value of combining aspirin and nutlin‑3 on hepatocellular carcinoma (HCC) cells. MTT was performed to detect the proliferation of HepG2 cells treated with aspirin or/and nutlin‑3. Transwell invasion assays were performed to estimate the invasion ability of HepG2 cells treated with aspirin or/and nutlin‑3. Then the apoptotic analysis of HepG2 cells evaluated the synergistic effect of aspirin and nutlin‑3. Apoptosis markers, including B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax), caspase‑3, caspase‑8 and caspase‑9 were estimated by western blot analysis at various time points. In addition, a Xenograft mouse model was established by infection with HepG2 cells, and aspirin and/or nutlin‑3 was administrated to verify the anti‑apoptotic effect of the two drugs in vivo. A high dose of aspirin and nutlin‑3 inhibit the proliferation and apoptosis of HepG2 cells. The antitumor effect was enhanced with the combined treatment of the two drugs, particularly in the group with a low concentration of aspirin and nutlin‑3. Nutlin‑3 was able to increase the level of Bax in HepG2 cells treated with aspirin significantly after treatment for 8 h. When treated with a low concentration of aspirin and nutlin‑3, the level of Bax in HepG2 cells was enhanced for 2 h. In the animal model, tumor volume and tumor angiogenesis were significantly decreased in combination group compared with other groups (P<0.01). Although there were side effects in the group treated with aspirin alone, no side effects were observed in the combination group. Nutlin‑3 enhanced the apoptotic effect of a low dose of aspirin by upregulating Bax expression in the HepG2 cell line and in vivo. The synergistic effect of nutlin‑3 in aspirin antitumor therapy contributed to diminishing the dose of aspirin required and decreased the occurrence of adverse drug events in HCC through targeting the Bcl‑2/Bax signaling pathway.
阿司匹林作为一种抗肿瘤药物，已在多种恶性肿瘤中进行了研究，涉及它对肿瘤细胞凋亡、增殖、转移和衰老的影响。然而，其副作用限制了它的临床应用。Nutlin-3 是一种新型抗肿瘤化合物，尚未在临床上获得批准。本研究探讨了阿司匹林和 Nutlin-3 联合应用于肝癌（HCC）细胞的价值。采用 MTT 法检测阿司匹林或/和 Nutlin-3 处理后 HepG2 细胞的增殖情况。采用 Transwell 侵袭实验评估阿司匹林或/和 Nutlin-3 处理后 HepG2 细胞的侵袭能力。然后通过流式细胞术分析 HepG2 细胞的凋亡，评估阿司匹林和 Nutlin-3 的协同作用。采用 Western blot 分析不同时间点的凋亡标志物，包括 B 细胞淋巴瘤 2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）、半胱天冬酶-3、半胱天冬酶-8 和半胱天冬酶-9。此外，通过 HepG2 细胞感染建立了异种移植小鼠模型，并给予阿司匹林和/或 Nutlin-3 以验证两种药物在体内的抗凋亡作用。高剂量的阿司匹林和 Nutlin-3 抑制 HepG2 细胞的增殖和凋亡。两种药物联合治疗可增强抗肿瘤作用，特别是在阿司匹林和 Nutlin-3 浓度较低的组中。Nutlin-3 可显著增加阿司匹林处理后 HepG2 细胞中 Bax 的水平，在处理 8 h 后达到峰值。当用低浓度的阿司匹林和 Nutlin-3 处理时，HepG2 细胞中 Bax 的水平在 2 h 时增强。在动物模型中，与其他组相比，联合组的肿瘤体积和肿瘤血管生成明显减少（P<0.01）。虽然单独使用阿司匹林组有副作用，但联合组没有观察到副作用。Nutlin-3 通过上调 HepG2 细胞系和体内 Bax 的表达增强了低剂量阿司匹林的凋亡作用。Nutlin-3 在阿司匹林抗肿瘤治疗中的协同作用通过靶向 Bcl-2/Bax 信号通路减少了所需的阿司匹林剂量，并降低了 HCC 中不良药物事件的发生。
