一种候选中药制剂——蔓荆子总黄酮可抑制肺癌干细胞样细胞的干细胞样特性。
A candidate Chinese medicine preparation-Fructus Viticis Total Flavonoids inhibits stem-like characteristics of lung cancer stem-like cells.
作者信息
Cao Xiaocheng, Zou Hui, Cao Jianguo, Cui Yinghong, Sun Shuwen, Ren Kaiqun, Song Zhenwei, Li Duo, Quan Meifang
机构信息
College of Life Science, Hunan Normal University, Changsha, China.
Laboratory of Medicine Engineering, College of Medicine, Hunan Normal University, Changsha, China.
出版信息
BMC Complement Altern Med. 2016 Sep 15;16:364. doi: 10.1186/s12906-016-1341-4.
BACKGROUND
Cancer stem cells (CSCs) are considered as the origin of tumor relapse. Here, we investigated the effects of Fructus Viticis total flavonoids (FVTF) on the characteristics of lung cancer stem-like cells (LCSLCs) derived from human small cell lung cancer NCI-H446 cell line and its potential mechanism.
METHODS
Human small cell lung cancer NCI-H446 cell line was cultured in vitro. The CD133(+) cells were sorted from NCI-H446 cell line by magnetic separation. The suspended culture with stem cell-conditioned medium was used to amplify CD133(+) sphere-forming cells (SFCs). The stem cell characteristics of CD133(+) SFCs were evaluated using cell self-renewal capacity by tumor sphere formation assay, migration and invasion capacity by Transwell assay, tumorigenicity by xenograft model in nude mouse and cancer stem cell markers expression levels by western blot. The effects of FVTF on the properties of LCSLCs were examined by tumorsphere formation assay and transwell chamber assay. The expression level of p-Akt was determined by western blot analysis.
RESULT
CD133(+) SFCs derived from human small cell lung cancer NCI-H446 cells exhibited stemness properties of tumorsphere formation and tumorigenesis capacity comparing to the parental cells. FVTF relative selectively inhibited the proliferation of LCSLCs, suppressed tumor sphere forming capacity and migration and invasion of LCSLCs, and down-regulated the protein expression of stem cell markers (CD133, CD44 and ALDH1), self-renewal associated transcription factors (Bmi1, Nanog and OCT4) and invasion associated transcription factors (Twist1 and Snail1) in a dose-dependent manner. Moreover, we found that FVTF treatment could significantly decrease the phosphorylation level of Akt in LCSLCs. Meanwhile, LY294002 and FVTF synergistically inhibited the characteristics of LCSLCs.
CONCLUSION
FVTF inhibits the characteristics of LCSLCs through down-regulating expression of p-Akt.
背景
癌症干细胞（CSCs）被认为是肿瘤复发的根源。在此，我们研究了蔓荆子总黄酮（FVTF）对源自人小细胞肺癌NCI-H446细胞系的肺癌干细胞样细胞（LCSLCs）特性的影响及其潜在机制。
方法
人小细胞肺癌NCI-H446细胞系在体外培养。通过磁珠分选从NCI-H446细胞系中分离出CD133(+)细胞。使用含干细胞条件培养基的悬浮培养来扩增CD133(+)成球细胞（SFCs）。通过肿瘤球形成试验评估CD133(+) SFCs的细胞自我更新能力，通过Transwell试验评估其迁移和侵袭能力，通过裸鼠异种移植模型评估其致瘤性，并通过蛋白质免疫印迹法检测癌症干细胞标志物的表达水平，以此来评价CD133(+) SFCs的干细胞特性。通过肿瘤球形成试验和Transwell小室试验检测FVTF对LCSLCs特性的影响。通过蛋白质免疫印迹分析确定p-Akt的表达水平。
结果
与人小细胞肺癌NCI-H446细胞系的亲代细胞相比，源自该细胞系的CD133(+) SFCs表现出肿瘤球形成和致瘤能力的干性特性。FVTF相对选择性地抑制LCSLCs的增殖，抑制其肿瘤球形成能力以及迁移和侵袭能力，并以剂量依赖性方式下调干细胞标志物（CD133、CD44和ALDH1）、自我更新相关转录因子（Bmi1、Nanog和OCT4）以及侵袭相关转录因子（Twist1和Snail1）的蛋白表达。此外，我们发现FVTF处理可显著降低LCSLCs中Akt的磷酸化水平。同时，LY294002和FVTF协同抑制LCSLCs的特性。
结论
FVTF通过下调p-Akt的表达来抑制LCSLCs的特性。
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