Enhanced anti-cancer effects of conditioned medium from hypoxic human adult dermal fibroblasts on cervical cancer cells [dataset 2]. Enhanced anti-cancer effects of conditioned medium from hypoxic human adult dermal fibroblasts on cervical cancer cells [dataset 2]

Hypoxia regulates fibroblast function by changing intracellular signaling and secretion factors, which influences the states of nearby cells. In this work, we investigated the effects of conditioned medium (CM) from human adult dermal fibroblasts (HDFs) cultured in normoxic and hypoxic conditions on cervical cancer (HeLa) cells. The HeLa cells showed decreased cell viability, in-creased apoptosis, and cell cycle arrest in response to CM from hypoxic-cultured HDFs (H-CM) compared with CM from normoxic-cultured HDFs (N-CM). Among the proteins up-regulated (> 2-fold) in H-CM compared with N-CM, LTBR decreased the viability of HeLa cells. Among the intracellular proteins down-regulated (> 2-fold) in HeLa cells treated with H-CM compared with N-CM, the most enriched biological process GO term and KEGG pathway were protein deubiq-uitination and hsa05166:HTLV-I infection, respectively. In the protein–protein interaction net-work of intracellular proteins with altered expression (> 2-fold), 1 up-regulated (TNF) and 8 down-regulated (ESR1, MCL1, TBP, CD19, LCK, PCNA, CHEK1, and POLA1) hub proteins were defined. Among the down-regulated hub proteins, the most enriched biological process GO term and KEGG pathway were leading strand elongation and hsa05166:HTLV-I infection, respectively. This study reveals that H-CM had enhanced anti-cancer effects on cervical cancer cells compared with N-CM, and it induced intracellular signaling patterns related to those enhanced anti-cancer effects. Overall design: Human adult dermal fibroblasts (HDFs) were cultured in 21% O2 as the normoxic condition and 1% O2 as the hypoxic condition. Conditioned medium from normoxic HDFs (N-CM) and hypoxic HDFs (H-CM) were harvested, and treated to HeLa cells for 24 hours. Up- and down-regulated intracellular proteins (> 2-fold) in HeLa cells treated with H-CM compared with N-CM were analyzed using Fullmoon Signaling explorer array.

缺氧可通过调控细胞内信号通路与分泌因子影响成纤维细胞功能，进而改变邻近细胞的状态。本研究针对常氧与缺氧条件下培养的成人真皮成纤维细胞（human adult dermal fibroblasts，HDFs）的条件培养基（conditioned medium，CM）对宫颈癌（HeLa）细胞的作用展开探究。结果显示，与常氧培养HDFs的条件培养基（N-CM）相比，缺氧培养HDFs的条件培养基（H-CM）处理后的HeLa细胞活力下降、凋亡水平升高，并出现细胞周期阻滞。
在H-CM相较于N-CM上调（>2倍）的蛋白中，LTBR可降低HeLa细胞的活力。在经H-CM相较于N-CM处理的HeLa细胞内下调（>2倍）的细胞内蛋白中，富集度最高的基因本体（Gene Ontology，GO）生物过程术语与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路分别为蛋白质去泛素化与hsa05166：人类T细胞白血病病毒I型（HTLV-I）感染。
在表达量改变（>2倍）的细胞内蛋白所构成的蛋白质相互作用网络中，共鉴定出1个上调核心蛋白（TNF）与8个下调核心蛋白（ESR1、MCL1、TBP、CD19、LCK、PCNA、CHEK1及POLA1）。在下调的核心蛋白中，富集度最高的GO生物过程术语与KEGG通路分别为前导链延伸与hsa05166：HTLV-I感染。
本研究证实，相较于N-CM，H-CM对宫颈癌细胞的抗肿瘤效应更强，并可诱导与该增强抗肿瘤效应相关的细胞内信号通路模式。
整体实验设计：将成人真皮成纤维细胞（HDFs）分别置于21% O₂的常氧环境与1% O₂的缺氧环境中培养。收集常氧HDFs的条件培养基（N-CM）与缺氧HDFs的条件培养基（H-CM），以其处理HeLa细胞24小时。采用Fullmoon信号探索芯片（Fullmoon Signaling explorer array）分析经H-CM相较于N-CM处理的HeLa细胞内上调与下调（>2倍）的蛋白。