Endothelial cells are the key components of vascular intima and play pivotal roles in vasculogenesis, angiogenesis, and tumor growth. Using Northern blot and real-time PCR, we confirmed that miR-126 and its host gene EGF-like domain 7 (EGFL7) were widely expressed in rat tissues but strictly expressed in endothelial cells. In mammals, miR-126 gene is embedded in intron7 of EGFL7. To explore the biogenesis of miR-126, plasmid EGFL7(126)-pEGFPc1 containing segment of exon7-intron7-exon8 of EGFL7 was constructed and expressed in 293T. Expression of spliced exon7-8 and excised mature miR-126 was detected by PCR and Northern blot. Knocking-down of endothelial endogenous miR-126 did not affect EGFL7 expression at mRNA or protein level. To investigate the possible roles of miR-126, PicTar, miRBase, miRanda, Bibiserv, and Targetscan were used to screen the targets. VEGFA and PIK3R2 were confirmed as the targets of miR-126 by luciferase reporter assay and Western blot. Interestingly, Northern blot and western blot showed that miR-126 was down-regulated in breast tumors where the VEGF/PI3K/AKT signaling pathway was activated. Introduction of miR-126 mimics into MCF-7 could effectively decrease VEGF/PI3K/AKT signaling activity. In summary, miR-126 was strictly expressed in endothelial cells and excised from EGFL7 pre-mRNA without affecting splicing and expression of its host gene. In addition, miR-126 could target both VEGFA and PIK3R2, and its expression was decreased in human breast cancer, implying that miR-126 may play a role in tumor genesis and growth by regulating the VEGF/PI3K/AKT signaling pathway.
Faced with the scarcity of water resource and irrational fertilizer use, it is highly important to supply plants with water and fertilizer at desiderated stages to improve yield with high water use efficiency (WUE). A pot experiment was conducted to investigate the effects of growth stage-specific water deficiency and potassium (K) fertilization on tomato yield and WUE. The entire growing season of tomato was divided into 5 stages: vegetative growth stage (VG), flowering and fruit setting stage (FS), early fruit growth stage (FG), fruit development stage (FD) and fruit maturity stage (FM). Three soil moisture (W) and three K fertilization levels were set up. W levels included W1, W2 and W3, indicating that soil water was maintained at 60–70% field capacity, 70–80% field capacity, and 80–90% field capacity, respectively. K levels included K1, K2 and K3, indicating that 0 g K
2
O per kg soil, 0.46 g K
2
O per kg soil and 0.92 g K
2
O per kg soil was applied. All combinations of the three W and three K levels were solely imposed at each of the five growth stages, for other four stages, plants were watered to 80–90% field capacity without K fertilizer (W3K1). The permanent W3K1 over the entire growth stage was taken as control (CK). The results showed that W deficiency imposed at all stages significantly affected tomato yield (P<0.01), except for VG stage in which W deficiency did not cause yield loss. K fertilization level during FS or FM stage had a significant effect on yield (P<0.01). A significant interaction effect of W and K on yield was only observed during FM stage. For WUE, significant effect of W deficiency at FS, FD and FM stages were observed, and a significant effect of K levels at FS, FD and FM stages was observed. Specifically, K fertilization was necessary during specific growth stage of tomato (i.e. FS and FM). During FS stage, even if a sufficient water supply seems necessary, a deficit irrigation with K fertilization could be applied as K fertilization could alleviate the negative effect of soil water deficit, however, excess of K fertilization during FM stage should be avoided to maintain tomato yield and WUE.
Effects of partial root-zone irrigation (PRI) on the hydraulic conductivity in the soil–root system (Lsr) in different root zones were investigated using a pot experiment. Maize plants were raised in split-root containers and irrigated on both halves of the container (conventional irrigation, CI), on one side only (fixed PRI, FPRI), or alternately on one of two sides (alternate PRI, APRI). Results show that crop water consumption was significantly correlated with Lsr in both the whole and irrigated root zones for all three irrigation methods but not with Lsr in the non-irrigated root zone of FPRI. The total Lsr in the irrigated root zone of two PRIs was increased by 49.0–92.0% compared with that in a half root zone of CI, suggesting that PRI has a significant compensatory effect of root water uptake. For CI, the contribution of Lsr in a half root zone to Lsr in the whole root zone was ∼50%. For FPRI, the Lsr in the irrigated root zone was close to that of the whole root zone. As for APRI, the Lsr in the irrigated root zone was greater than that of the non-irrigated root zone. In comparison, the Lsr in the non-irrigated root zone of APRI was much higher than that in the dried zone of FPRI. The Lsr in both the whole and irrigated root zones was linearly correlated with soil moisture in the irrigated root zone for all three irrigation methods. For the two PRI treatments, total water uptake by plants was largely determined by the soil water in the irrigated root zone. Nevertheless, the non-irrigated root zone under APRI also contributed to part of the total crop water uptake, but the continuously non-irrigated root zone under FPRI gradually ceased to contribute to crop water uptake, suggesting that it is the APRI that can make use of all the root system for water uptake, resulting in higher water use efficiency.
Osteosarcoma (OS) is among the most frequently occurring bone tumors, particularly in children. Clinical treatment of OS is limited due to several factors including resistance to chemotherapy drugs and metastasis, and the underlying molecular mechanisms remain unclear. In the present study, tripartite motif containing 37 (TRIM37) expression levels were upregulated in tumor samples and associated with the development of drug resistance in OS. Furthermore, chemotherapy drug treatment (doxorubicin, cisplatin and methotrexate) induced TRIM37 expression in OS cells in vitro. TRIM37 mRNA and protein were upregulated in 41 pediatric osteosarcoma clinical specimens. To further elucidate the effect of TRIM37, gain and loss-of-function analysis was performed. Overexpression of TRIM37 induced cell proliferation and drug resistance ability of OS cells, whilst TRIM37 knockdown suppressed cell growth rate and restored chemosensitivity. TRIM37-regulated genes were subsequently analyzed by expression microarray and gene set enrichment analysis. Using the Wnt/β-catenin inhibitor XAV-939, the present study demonstrated that TRIM37-induced chemoresistance is partially dependent on the activation of the Wnt/β-catenin signaling pathway. Collectively, the results of the present study suggest that TRIM37 may have a key role in the development of OS and in the ability for the cells to acquire drug resistance, thus it may be a novel target for the treatment of OS.
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