Department of Integrative Medical Biology, Umeå University, Umeå, Sweden.
Department of Chemistry, Umeå University, Umeå, Sweden.
J Cell Biochem. 2018 Feb;119(2):2320-2332. doi: 10.1002/jcb.26394. Epub 2017 Oct 18.
The cell-based therapies could be potential methods to treat damaged cartilage tissues. Instead of native hyaline cartilage, the current therapies generate mainly weaker fibrocartilage-type of repair tissue. A correct microenvironment influences the cellular phenotype, and together with external factors it can be used, for example, to aid the differentiation of mesenchymal stem cells to defined types of differentiated adult cells. In this study, we investigated the effect of long-term exposure to 5% low oxygen atmosphere on human chondrosarcoma HCS-2/8 cells. This atmosphere is close to normal oxygen tension of cartilage tissue. The proteome was analyzed with label-free mass spectrometric method and further bioinformatic analysis. The qRT-PCR method was used to gene expression analysis, and ELISA and dimethylmethylene blue assays for type II collagen and sulfated glycosaminoglycan measurements. The 5% oxygen atmosphere did not influence cell proliferation, but enhanced slightly ACAN and COL2A1 gene expression. Proteomic screening revealed a number of low oxygen-induced protein level responses. Increased ones included NDUFA4L2, P4HA1, NDRG1, MIF, LDHA, PYGL, while TXNRD1, BAG2, TXN2, AQSTM1, TNFRSF1B, and EPHX1 decreased during the long-term low oxygen atmosphere. Also a number of proteins previously not related to low oxygen tension changed during the treatment. Of those S100P, RPSS26, NDUFB11, CDV3, and TUBB8 had elevated levels, while ALCAM, HLA-B, EIF1, and ACOT9 were lower in the samples cultured at low oxygen tension. In conclusion, low oxygen condition causes changes in the cellular amounts of several proteins.
细胞疗法可能是治疗受损软骨组织的潜在方法。与天然透明软骨不同，目前的治疗方法主要产生较弱的纤维软骨样修复组织。正确的微环境会影响细胞表型，与外部因素一起，可以用于例如，辅助间充质干细胞向特定类型的分化成年细胞分化。在这项研究中，我们研究了长期暴露于 5%低氧环境对人软骨肉瘤 HCS-2/8 细胞的影响。这种气氛接近于软骨组织的正常氧张力。使用无标记质谱法和进一步的生物信息学分析分析蛋白质组。使用 qRT-PCR 方法进行基因表达分析，以及 ELISA 和二甲亚甲基蓝测定法测定 II 型胶原和硫酸化糖胺聚糖。5%的氧气环境不会影响细胞增殖，但略微增强了 ACAN 和 COL2A1 基因的表达。蛋白质组筛选揭示了许多低氧诱导的蛋白质水平反应。增加的包括 NDUFA4L2、P4HA1、NDRG1、MIF、LDHA、PYGL，而 TXNRD1、BAG2、TXN2、AQSTM1、TNFRSF1B 和 EPHX1 在长期低氧环境下减少。此外，在治疗过程中，一些以前与低氧张力无关的蛋白质也发生了变化。其中 S100P、RPSS26、NDUFB11、CDV3 和 TUBB8 的水平升高，而 ALCAM、HLA-B、EIF1 和 ACOT9 在低氧张力培养的样品中水平降低。总之，低氧条件会导致细胞内多种蛋白质的含量发生变化。
