亚硝酸盐调控人肝癌SMMC-7721细胞癌干细胞特性

发布时间：2018-10-19 12:15

【摘要】：有证据表明肝癌含有一定数量的具有自我更新能力的癌干细胞亚群。然而,癌干细胞是如何被调控的还不清楚。肿瘤微环境中一些生物介质可以导致癌干细胞快速可逆性改变。本文旨在探讨,在肿瘤微环境中有较高含量,又是细胞关键的调控因子的亚硝酸盐是如何调控人肝癌干细胞样细胞表型的。用SMMC-7721细胞系无血清培养获得细胞球。流式细胞术检测细胞周期,体外克隆形成实验和球形成实验检测细胞自我更新能力,侵袭实验检测细胞侵袭能力,癌干细胞标志物检测采用流式细胞术和Westen blot方法,MTT检测细胞活力。皮下注射SMMC-7721球形成细胞建立裸鼠移植瘤模型,活性氧(ROS)检测试剂盒检测移植瘤组织内ROS水平,免疫荧光检测组织内缺氧有诱导因子-1α(HIF-1α)。结果显示,SMMC-7721细胞系来源的球细胞自我更新能力增强,干细胞标志物表达增加,耐药性增强,提示球形成细胞富集了较多的具有癌干细胞特征的细胞。用亚硝酸钠(150μmol·L~(-1))处理亲本细胞和球形成细胞后,G_0/G_1期细胞比例增加,肝癌干细胞标志物CD133、CD90和EpCAM表达增加,细胞耐药能力和侵袭能力增强。与亲本细胞相比,亚硝酸盐对球形成细胞的上述作用明显增强。体内实验结果也显示,亚硝酸盐促进球形成细胞移植瘤生长,组织内ROS水平下降,HIF-1α累积。结果表明,亚硝酸盐通过上调肝癌干细胞"干性"增强肝癌细胞侵袭能力。
[Abstract]:There is evidence that liver cancer contains a certain number of self-renewing cancer stem cell subsets. However, it is unclear how cancer stem cells are regulated. Some biological mediators in tumor microenvironment can cause rapid reversible changes of cancer stem cells. The aim of this paper is to investigate how nitrite, which is a key regulatory factor and a high content of tumor microenvironment, regulates the phenotype of human hepatoma stem cell like cells. The cell spheres were obtained by serum-free culture of SMMC-7721 cell line. The cell cycle was detected by flow cytometry, the ability of cell self-renewal was detected by colony formation in vitro and the ability of cell invasion was detected by invasiveness assay. Cancer stem cell markers were detected by flow cytometry and Westen blot, and MTT was used to detect cell viability. The xenograft tumor model of nude mice was established by subcutaneous injection of SMMC-7721 ball forming cells. Reactive oxygen species (Ros) (ROS) assay kit was used to detect the level of ROS in transplanted tumor tissues. Immunofluorescence was used to detect hypoxia inducible factor-1 伪 (HIF-1 伪). The results showed that the spherical cells derived from SMMC-7721 cell lines had enhanced self-renewal ability, increased expression of stem cell markers and increased drug resistance, which suggested that more cells with the characteristics of cancer stem cells were enriched in the globe-forming cells. After treated with sodium nitrite (150 渭 mol L ~ (-1), the proportion of G_0/G_1 phase cells increased, the expression of CD133,CD90 and EpCAM, and the ability of cell resistance and invasion increased. Compared with parental cells, nitrite significantly enhanced the effect of nitrite on globular cells. The results of in vivo experiments also showed that nitrite promoted the growth of granuloma, decreased the level of ROS and accumulated HIF-1 伪 in tissues. The results showed that nitrite enhanced the invasion ability of hepatoma cells by up-regulating the "dryness" of hepatoma stem cells.
【作者单位】：河南大学护理学院;河南大学第一附属医院;河南大学医学院环境医学研究所;
【基金】：国家环保公益项目专项(200809115) 省部共建河南大学科研项目(SBGJ090702)
【分类号】：R735.7

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