Exosome‐derived miRNAs are regarded as biomarkers for the diagnosis and prognosis of many human cancers. However, its function in clear cell renal cell carcinoma (ccRCC) remains unclear. In this study, differentially expressed miRNAs from urinal exosomes were identified using next‐generation sequencing (NGS) and verified using urine samples of ccRCC patients and healthy donors. Then, the exosomes were analysed in early‐stage ccRCC patients, healthy individuals and patients suffering from other urinary system cancers. Thereafter, the target gene of the miRNA was detected. Its biological function was investigated in vitro and in vivo. The results showed that miR‐30c‐5p could be amplified in a stable manner. Its expression pattern was significantly different only between ccRCC patients and healthy control individuals, but not compared with that of other urinary system cancers, which indicated its specificity for ccRCC. Additionally, the overexpression of miR‐30c‐5p inhibited ccRCC progression in vitro and in vivo. Heat‐shock protein 5 (HSPA5) was found to be a direct target gene of miR‐30c‐5p. The depletion of HSPA5 caused by miR‐30c‐5p inhibition reversed the promoting effect of ccRCC growth. In conclusion, urinary exosomal miR‐30c‐5p acts as a potential diagnostic biomarker of early‐stage ccRCC and may be able to modulate the expression of HSPA5, which is correlated with the progression of ccRCC.
Background The heterotrimeric G protein β subunit RGB1 plays an important role in plant growth and development. However, the molecular mechanisms underlying the regulation of rice growth by RGB1 remain elusive. Results Here, the rgb1 mutants rgb1–1 (+ 1 bp), rgb1–2 (− 1 bp), and rgb1–3 (− 11 bp) were isolated using the CRISPR/Cas9 system, and they were arrested at 1 day after germination and ultimately exhibited seedling lethality. The dynamic anatomical characteristics of the embryos of the rgb1 seedlings and WT during early postgermination and according to TUNEL assays showed that the suppressed growth of the rgb1 mutants was caused by cell death. In addition to the limited shoot and root development, the development of the embryo shoot-root axis was suppressed in the rgb1 mutants. RGB1 was expressed mainly in the root epidermal and vascular tissues of the embryo. Moreover, transcript profiling analysis revealed that the expression of a large number of auxin-, cytokinin-, and brassinosteroid-inducible genes was upregulated or downregulated in the rgb1 mutant compared to the wild type during seedling development. Conclusions Overall, the rgb1 mutants provide an ideal material for exploring the molecular mechanism underlying rice seedling formation during early postgermination development by G proteins. Significance statement The heterotrimeric G protein β subunit RGB1 acts as a crucial factor in promoting early postgermination seedling development in rice. Electronic supplementary material The online version of this article (10.1186/s12284-019-0313-y) contains supplementary material, which is available to authorized users.
Background Brassinosteroids (BRs) are a new group of plant hormones and play important roles in plant growth and development. However, little information is available if BRs could regulate spikelet development in rice (Oryza sativa L.) especially under soil-drying conditions. This study investigated whether and how BRs mediate the effect of soil-drying on spikelet differentiation and degeneration in rice. A rice cultivar was field-grown and exposed to three soil moisture treatments during panicle development, that is, well-watered (WW), moderate soil-drying (MD) and severe soil-drying (SD). Results Compared with the WW treatment, the MD treatment enhanced BRs biosynthesis in young panicles, increased spikelet differentiation and reduced spikelet degeneration. The SD treatment had the opposite effects. Changes in expression levels of key rice inflorescence development genes (OsAPO2 and OsTAW1), ascorbic acid (AsA) content, and activities of enzymes involved AsA synthesis and recycle, and amount of nonstructural carbohydrates (NSC) in young panicles were consistent with those in BRs levels, whereas hydrogen peroxide (H2O2) content showed opposite trend. Knockdown of the BRs synthesis gene OsD11 or application of a BRs biosynthesis inhibitor to young panicles markedly decreased OsAPO2 and OsTAW1 expression levels, BRs and AsA contents, activities of enzymes involved AsA synthesis and recycle, NSC amount in rice panicles and spikelet differentiation but increased the H2O2 content and spikelet degeneration compared to the control (the wide type or application of water). The opposite effects were observed when exogenous BRs were applied. Conclusions The results suggest that BRs mediate the effect of soil-drying on spikelet differentiation and degeneration, and elevated BRs levels in rice panicles promote spikelet development under MD by enhancing inflorescence meristem activity, AsA recycle and NSC partitioning to the growing panicles.
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