Gomes Sofia E, Pereira Diane M, Roma-Rodrigues Catarina, Fernandes Alexandra R, Borralho Pedro M, Rodrigues Cecília M P
Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Lisbon, Portugal.
UCIBIO, Departamento de Ciências da Vida, Faculty of Sciences and Technology, New University of Lisbon, Caparica, Portugal.
PLoS One. 2018 Jan 23;13(1):e0191607. doi: 10.1371/journal.pone.0191607. eCollection 2018.
MicroRNAs (miRNAs) regulate a wide variety of biological processes, including tumourigenesis. Altered miRNA expression is associated with deregulation of signalling pathways, which in turn cause abnormal cell growth and de-differentiation, contributing to cancer. miR-143 and miR-145 are anti-tumourigenic and influence the sensitivity of tumour cells to chemotherapy and targeted therapy. Comparative proteomic analysis was performed in HCT116 human colon cancer cells stably transduced with miR-143 or miR-145. Immunoblotting analysis validated the proteomic data in stable and transient miRNA overexpression conditions in human colon cancer cells. We show that approximately 100 proteins are differentially expressed in HCT116 human colon cancer cells stably transduced with miR-143 or miR-145 compared to Empty control cells. Further, Gene Ontology and pathway enrichment analysis indicated that proteins involved in specific cell signalling pathways such as cell death, response to oxidative stress, and protein folding might be modulated by these miRNAs. In particular, antioxidant enzyme superoxide dismutase 1 (SOD1) was downregulated by stable expression of either miR-143 or miR-145. Further, SOD1 gain-of-function experiments rescued cells from miR-143-induced oxidative stress. Moreover, miR-143 overexpression increased oxaliplatin-induced apoptosis associated with reactive oxygen species generation, which was abrogated by genetic and pharmacological inhibition of oxidative stress. Overall, miR-143 might circumvent resistance of colon cancer cells to oxaliplatin via increased oxidative stress in HCT116 human colon cancer cells.
微小RNA（miRNA）调节多种生物学过程，包括肿瘤发生。miRNA表达的改变与信号通路失调相关，进而导致细胞异常生长和去分化，促进癌症发生。miR-143和miR-145具有抗肿瘤作用，并影响肿瘤细胞对化疗和靶向治疗的敏感性。对稳定转导miR-143或miR-145的HCT116人结肠癌细胞进行了比较蛋白质组学分析。免疫印迹分析验证了人结肠癌细胞在稳定和瞬时miRNA过表达条件下的蛋白质组学数据。我们发现，与空载体对照细胞相比，稳定转导miR-143或miR-145的HCT116人结肠癌细胞中约有100种蛋白质表达存在差异。此外，基因本体论和通路富集分析表明，参与特定细胞信号通路（如细胞死亡、氧化应激反应和蛋白质折叠）的蛋白质可能受这些miRNA调控。特别是，抗氧化酶超氧化物歧化酶1（SOD1）在稳定表达miR-143或miR-145时被下调。此外，SOD1功能获得实验使细胞从miR-143诱导的氧化应激中恢复。此外，miR-143过表达增加了奥沙利铂诱导的与活性氧生成相关的细胞凋亡，而氧化应激的基因和药理学抑制可消除这种凋亡。总体而言，miR-143可能通过增加HCT116人结肠癌细胞中的氧化应激来规避结肠癌细胞对奥沙利铂的耐药性。
