Apoptosis in human liver carcinoma caused by gold nanoparticles in combination with carvedilol is mediated via modulation of MAPK/Akt/mTOR pathway and EGFR/FAAD proteins
Apoptosis in human liver carcinoma caused by gold nanoparticles in combination with carvedilol is mediated via modulation of MAPK/Akt/mTOR pathway and EGFR/FAAD proteins
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DOI：
10.3892/ijo.2017.4179
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发表时间：
2018-01-01
影响因子：
5.2
通讯作者：
Araujo, Aurigena A.
中科院分区：
文献类型：
--
作者：
De Araujo, Raimundo F., Jr.;Pessoa, Jonas B.;Araujo, Aurigena A.
在癌症中，导致对PI3K/Akt/mTOR和丝裂原活化蛋白激酶(MAPK)等常规治疗耐药的细胞凋亡信号通路和细胞生存和生长通路变得失调。最近，促进肿瘤细胞死亡的替代疗法变得重要起来。本研究报道了金纳米粒(GNPs)与卡维地洛联合及单独应用的抗肿瘤和细胞保护作用。流式细胞仪检测细胞凋亡，免疫荧光显微镜检测caspase-3、caspase-8、bcl2和MAPK/ERK活性，实时定量聚合酶链式反应检测与细胞死亡相关的蛋白Akt、mTOR、EGFR、mdr1、Survivin、FADD和APAF的基因表达，Western印迹分析MAPK/ERK、Akt和mTOR。通过还原型谷胱甘肽(GSH)和丙二醛(MDA)水平评估氧化应激。用透射电子显微镜鉴定细胞内的GNPs靶点。GNPs(6.25mU/ml)和卡维地洛(3mU M)联合作用后，流式细胞仪检测促凋亡蛋白FADD、caspase-3、caspase-8的表达以及对抗凋亡的MAPK/ERK、Akt、mTOR、EGFR和mdr1耐药的亚调节作用。检测GSH升高和丙二醛水平降低的非肿瘤细胞细胞保护作用。当与卡维地洛结合时，在靠近细胞核的细胞内发现了GNP。GNP和卡维地洛联合应用可促进肿瘤细胞中抗凋亡和耐药基因的下调，促进促凋亡蛋白的过度表达，并通过减少细胞凋亡和氧化应激来保护非肿瘤细胞。
In cancers, apoptosis signaling pathways and cell survival and growth pathways responsible for resistance to conventional treatments, such as Pi3K/Akt/mTOR and mitogen-activated protein kinase (MAPK) become dysregulated. Recently, alternative treatments to promote tumor cell death have become important. The present study reports on the antitumor and cytoprotective action of gold nanoparticles (GNPs) and carvedilol in combination and in isolated application. Apoptosis was analyzed by FITC/propidium iodide staining flow cytometry; caspase-3, caspase-8, Bcl-2 and MAPK/ERK activity by immunofluorescence microscopy; gene expression of proteins related to cell death as Akt, mTOR, EGFR, MDR1, survivin, FADD and Apaf, by the real-time PCR; and western blot analysis for MAPK/ERK, Akt and mTOR. Oxidative stress evaluation was performed by reduced glutathione (GSH) and malondialdehyde (MDA) levels. Intracellular GNPs targets were identified by transmission electron microscopy. After exposure to a combination of GNPs (6.25 mu g/ml) and carvedilol (3 mu M), death as promoted by apoptosis was detected using flow cytometry, for expression of pro-apoptotic proteins FADD, caspase-3, caspase-8 and sub-regulation of anti-apoptotic MAPK/ERK, Akt, mTOR, EGFR and MDR1 resistance. Non-tumor cell cytoprotection with GSH elevation and MDA reduction levels was detected. GNPs were identified within the cell near to the nucleus when combined with carvedilol. The combination of GNP and carvedilol promoted downregulation of anti-apoptotic and drug resistance genes, over-regulation of pro-apoptotic proteins in tumor cells, as well as cytoprotection of non-tumor cells with reduction of apoptosis and oxidative stress.