ABSTRACT
The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G(0)-G(1) phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST-PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.
Subject(s)
Bronchi/cytology , Epithelial Cells/cytology , Gene Expression Regulation, Neoplastic , MicroRNAs/metabolism , Simian virus 40/metabolism , Up-Regulation , Animals , Antigens, Viral, Tumor/metabolism , Cell Line, Tumor , Cell Proliferation , Cell Transformation, Neoplastic , Epigenesis, Genetic , Humans , Mice , Mice, SCID , Signal TransductionABSTRACT
A regulator of the protein phosphatase 2A (PP2A), α4, has been implicated in a variety of functions that regulate many cellular processes. To explore the role of α4 in human cell transformation and tumorigenesis, we show that α4 is highly expressed in human cells transformed by chemical carcinogens including benzo(a)pyrene, aflatoxin B(1), N-methyl-N'-nitro-N-nitrosoguanidine, nickel sulfate and in several hepatic and lung cancer cell lines. In addition, overexpression of α4 was detected in 87.5% (74/80) of primary hepatocellular carcinomas, 84.0% (21/25) of primary lung cancers and 81.8% (9/11) of primary breast cancers, indicating that α4 is ubiquitously highly expressed in human cancer. Functional studies revealed that elevated α4 expression results in an increase in cell proliferation, promotion of cell survival and decreased PP2A-attributable activity. Importantly, ectopic expression of α4 permits non-transformed human embryonic kidney cells (HEKTER) and L02R cells to form tumors in immunodeficient mice. Furthermore, we show that the highly expressed α4 in transformed cells or human tumors is not regulated by DNA hypomethylation. A microRNA, miR-34b, that suppresses the expression of α4 through specific binding to the 3'-untranslated region of α4 is downregulated in transformed or human lung tumors. Taken together, these observations identify that α4 possesses an oncogenic function. Reduction of PP2A activity due to an enhanced α4-PP2A interaction contributes directly to chemical carcinogen-induced tumorigenesis.
Subject(s)
Cell Transformation, Neoplastic/genetics , Intracellular Signaling Peptides and Proteins/genetics , Neoplasms/genetics , Adaptor Proteins, Signal Transducing , Animals , Base Sequence , Carcinogens , Cell Line, Transformed , Cell Transformation, Neoplastic/chemically induced , Cell Transformation, Neoplastic/drug effects , Cell Transformation, Neoplastic/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Gene Knockdown Techniques , Hep G2 Cells , Humans , Intracellular Signaling Peptides and Proteins/metabolism , Intracellular Signaling Peptides and Proteins/physiology , Mice , Mice, Inbred BALB C , Mice, Nude , MicroRNAs/physiology , Molecular Chaperones , Neoplasms/chemically induced , RNA, Small Interfering/pharmacology , Transplantation, Heterologous , Tumor Cells, Cultured , Up-RegulationABSTRACT
The effects of instilled silica have been studied on the serum-phospholipid (PL), lipid peroxide (LPO) and histopathology of rat lung up to 140 days from the first day of instillation. Silica induced relatively higher serum-PL throughout the experiment. The level of LPO also increased appreciably. They presented positive linear correlation. The early lesion was acute alveolitis with silica particles. These lesions became silicotic nodules on the 30th day, which then were enlarged gradually and fused by fibrosis. Alveolar macrophages (AM) were activated and surface structure was damaged. These results indicate that instilled silica can induce lipid peroxidation of cell membrane and selective accumulation of lung PL.
