Transposon insertional mutagenesis in mice identifies human breast cancer susceptibility genes and signatures for stratification.

Robust prognostic gene signatures and therapeutic targets are difficult to derive from expression profiling because of the significant heterogeneity within breast cancer (BC) subtypes. Here, we performed forward genetic screening in mice using Sleeping Beauty transposon mutagenesis to identify candidate BC driver genes in an unbiased manner, using a stabilized N-terminal truncated ß-catenin gene as a sensitizer. We identified 134 mouse susceptibility genes from 129 common insertion sites within 34 mammary tumors. Of these, 126 genes were orthologous to protein-coding genes in the human genome (hereafter, human BC susceptibility genes, hBCSGs), 70% of which are previously reported cancer-associated genes, and ∼16% are known BC suppressor genes. Network analysis revealed a gene hub consisting of E1A binding protein P300 (EP300), CD44 molecule (CD44), neurofibromin (NF1) and phosphatase and tensin homolog (PTEN), which are linked to a significant number of mutated hBCSGs. From our survival prediction analysis of the expression of human BC genes in 2,333 BC cases, we isolated a six-gene-pair classifier that stratifies BC patients with high confidence into prognostically distinct low-, moderate-, and high-risk subgroups. Furthermore, we proposed prognostic classifiers identifying three basal and three claudin-low tumor subgroups. Intriguingly, our hBCSGs are mostly unrelated to cell cycle/mitosis genes and are distinct from the prognostic signatures currently used for stratifying BC patients. Our findings illustrate the strength and validity of integrating functional mutagenesis screens in mice with human cancer transcriptomic data to identify highly prognostic BC subtyping biomarkers.

Allelopathic effects of volatile organic compounds from Eucalyptus grandis rhizosphere soil on Eisenia fetida assessed using avoidance bioassays, enzyme activity, and comet assays.

Allelopathy has been identified as an underlying mechanism of detrimental environmental impacts within commercial plantations. Eucalyptus spp. are known to generate huge amounts of volatile organic compounds (VOCs) that can function as phytotoxins and thus inhibit other plants. In the present study, biochemical markers, including activities of acetylcholinesterase (AChE) and oxidative stress enzymes, such as superoxide dismutase (SOD) and glutathione S-transferase (GST), were assayed to assess changes in Eisenia fetida at the physiological level induced by different doses of VOCs as part of an acute toxicity test over 7 and 14-day exposures. In addition, the toxicities of VOCs were investigated using a soil avoidance test and comet assay. The results revealed that E. fetida exhibited significant avoidance behavior towards the highest concentrations of undecane, decane, 2,4-dimethyl heptane, and 2,2,4,6,6-pentametyl heptane. The tail DNA percentages were significantly increased for all experimental treatments relative to control. However, under the treatments of VOCs, Olive tail moment content and comet tail length also display an obvious increase compared to control, except for that of octane, undecane and decane treatments. As VOC concentrations and durations increased in the soil, activities of AChE, SOD, and GST were either stimulated or inhibited. Among the VOCs, decane, 2,4-dimethyl heptane, 2,2,4,6,6-pentamethyl heptane, and 2,4-di tert buyl phenol exerted stronger effects on enzymatic activities. In summary, VOCs in rhizosphere soils of E. grandis might exert a toxic impact on E. fetida, among which 2,4-dimethyl heptane, 2,2,4,6,6-pentamethyl heptane, and 2,4-di tert buyl phenol have the strongest effects.