Survival advantages of multicellular spheroids vs. monolayers of HepG2 cells in vitro
Survival advantages of multicellular spheroids vs. monolayers of HepG2 cells in vitro
复制标题
DOI：
10.3892/or_00000167
复制
发表时间：
2008-12-01
期刊：
影响因子：
4.2
通讯作者：
Zhang, Wang-Gang
中科院分区：
文献类型：
--
作者：
Li, Chun-Li;Tian, Tao;Zhang, Wang-Gang
哺乳动物细胞在体内以三维（3-D）生长。常用的二维（2-D）细胞培养不足以重建肿瘤细胞的生物微环境。2-D和3-D培养系统的潜在不同结果可能对实验结果的相关性产生重大影响。本研究的目的是在2-D和3-D培养中表征人肝癌细胞系HepG 2。2-D和3-D培养的HepG 2细胞用顺铂、5-氟尿嘧啶和阿霉素处理，并通过扫描电子显微镜和透射电子显微镜分析。流式细胞仪检测细胞周期进程和凋亡。通过MTT测定定量细胞增殖的抑制。免疫组化染色检测E-cadherin、CD 44 v6、VEGF、KDR、endostatin、Bax和cytochrome-c的表达。与二维单层培养相比，三维多细胞球体（MCS）的HepG 2细胞具有更大比例的细胞处于G1期，并具有更丰富的细胞间粘附。此外，MCS中的细胞在维持培养基中凋亡显著较少，并且对药物诱导的凋亡更具抗性。与2-D细胞培养物相比，MCS中的E-钙粘蛋白、CD 44 v6、VEGF、KDR、内皮抑素和细胞色素c水平增加。结论：MCS可诱导不同的表型，包括细胞间粘附增加、G1期细胞阻滞、细胞抗凋亡能力增强和血管生成能力上调。基于我们的数据，多细胞形态层次可能维持癌细胞在体内的生长/存活优势。因此，三维培养系统应是肿瘤生物学研究的首选技术。
Mammalian cells grow in three-dimensions (3-D) in vivo. Commonly used two-dimensional (2-D) cell Cultures are inadequate to recreate the biological microenvironment of tumor cells. The potentially different outcomes from 2-D and 3-D culture systems may have a significant impact oil the relevance of experimental findings. The purpose of this study was to characterize the human hepatoma cell line HepG2 in 2-D and 3-D Cultures. HepG2 cells in 2-D and 3-D cultures were treated with cisplatin, 5-fluorouracil, and adriamycin and were analyzed by scanning electron microscopy and transmission electron microscopy. Cell cycle progression and apoptosis were detected by flow cytometry. Inhibition of cell proliferation was quantified by MTT assay. The expression of E-cadherin, CD44v6, VEGF, KDR, endostatin, Bax, and cytochrome-c were analyzed by immunohistochemical (IHC) staining. As compared to the 2-D monolayer culture, the 3-D multicellular spheroids (MCS) of HepG2 cells featured a greater fraction of cells in G I phase and were organized with more abundant cell-cell adhesion. In addition, cells in MCS were significantly less apoptotic in maintenance culture media and were more resistant to drug-induced apoptosis. E-cadherin, CD44v6, VEGF, KDR, endostatin, and cytochrome-c levels were increased in MCS as compared to 2-D cell cultures. In coclusion, MCS conferred differ-entiated phenotypes including increased cell-cell adhesion and G1 phase cell cycle arrest, enhanced cellular resistance to apoptosis, and upregulated angiogenic potential. Based on our data, a multicellular morphological hierarchy may sustain the growth/survival advantages of cancer cells in vivo. Therefore, a 3-D culture system should be the preferred technique for cancer biology investigation.