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苦参碱对从人肝癌SMMC - 7721细胞系分离出的癌干细胞样细胞的抗肿瘤作用。

Antitumor effects of matrine on cancer stem like cells isolated from the human liver cancer SMMC-7721 cell line.

作者信息

Wang Yong, Liu Yahui, Jiang Jianshuai, Cui Hanbin

机构信息

Key Lab of Ningbo, Ningbo First Hospital, Ningbo, Zhejiang 315010, P.R. China.

出版信息

Oncol Lett. 2018 Feb;15(2):1777-1782. doi: 10.3892/ol.2017.7531. Epub 2017 Dec 5.

Abstract

The existence of cancer stem cells (CSCs) or cancer stem-like cells (CSLCs) is regarded as the cause of tumor formation and recurrence. Matrine has been reported to exhibit antitumor effects in cancer cells. In the present study, a preliminary study was performed on the mechanisms of matrine on hepatocellular carcinoma (HCC) stem-like cells. The HCC SMMC-7721 cell line was cultured in tumor stem cell-specific medium to form spheres, and different concentrations (1, 2 and 5 mg/kg) of cisplatin were then used in order to purify the most drug-resistant cells, which were used as CSLCs. An MTT assay was performed to detect the inhibitory effects of matrine against CSLC proliferation. Quantitative polymerase chain reaction (qPCR) and western blot analysis were used to detect changes in cell adhesion regulating gene (CAR), E-cadherin, laminin and fibronectin. As a result, using tryptose sulfite cycloserine medium culture and cisplatin-resistance screening, CSLCs were successfully isolated from the SMMC-7721 cell line. Matrine inhibited the proliferation of CSLCs . The results of qPCR and western blot analysis demonstrated that matrine upregulated the expression of CAR, E-cadherin, laminin and fibronectin in CSLCs compared with the control treatment. A certain concentration of matrine exhibited antitumor effects on HCC stem like cells.

摘要

癌症干细胞(CSCs)或癌症干细胞样细胞(CSLCs)的存在被认为是肿瘤形成和复发的原因。据报道,苦参碱对癌细胞具有抗肿瘤作用。在本研究中,对苦参碱作用于肝癌(HCC)干细胞样细胞的机制进行了初步研究。将HCC SMMC - 7721细胞系在肿瘤干细胞特异性培养基中培养以形成球体,然后使用不同浓度(1、2和5 mg/kg)的顺铂来纯化耐药性最强的细胞,这些细胞用作CSLCs。进行MTT试验以检测苦参碱对CSLC增殖的抑制作用。采用定量聚合酶链反应(qPCR)和蛋白质印迹分析来检测细胞黏附调节基因(CAR)、E - 钙黏蛋白、层粘连蛋白和纤连蛋白的变化。结果,通过胰蛋白胨亚硫酸盐环丝氨酸培养基培养和顺铂耐药性筛选,成功从SMMC - 7721细胞系中分离出CSLCs。苦参碱抑制了CSLCs的增殖。qPCR和蛋白质印迹分析结果表明,与对照处理相比,苦参碱上调了CSLCs中CAR、E - 钙黏蛋白、层粘连蛋白和纤连蛋白的表达。一定浓度的苦参碱对HCC干细胞样细胞具有抗肿瘤作用。

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