Phosphorylation-dependent deubiquitinase OTUD3 regulates YY1 stability and promotes colorectal cancer progression.

Yin Yang 1 (YY1) is a key transcription factor that has been implicated in the development of several malignancies. The stability of YY1 is regulated by the ubiquitin-proteasome system. The role of deubiquitinases (DUBs) and their impact on YY1 remain to be fully elucidated. In this study, we screened for ubiquitin-specific proteases that interact with YY1, and identified OTUD3 as a DUB for YY1. Over-expressed OTUD3 inhibited YY1 degradation, thereby increasing YY1 protein levels, whereas OTUD3 knockdown or knockout promoted YY1 degradation, thereby decreasing the proliferation of colorectal cancer (CRC). Furthermore, PLK1 mediates OTUD3 S326 phosphorylation, which further enhances OTUD3 binding and deubiquitination of YY1. In CRC tissues, elevated the expression level of OTUD3 and YY1 were significantly associated with poor prognostic outcomes. These findings suggest that the OTUD3-YY1 pathway has therapeutic potential in CRC, and OTUD3 plays a critical role in regulating YY1.

LncRNA CASC15 regulates breast cancer cell stemness via the miR-654-5p/MEF2D axis.

Emerging evidence has demonstrated the prognostic and diagnostic potential of long noncoding RNA (lncRNA) cancer susceptibility candidate 15 (lncRNA CASC15) for the progression and tumorigenesis of human cancer. However, how CASC15 modulates the stemness of breast cancer stem cells (BCSCs) is not well understood. In this study, high expression of CASC15 in MCF-7 CSCs was reported, relative to MCF-7 cells, and this phenomenon was associated with metastatic lymph nodes, higher TNM stage, and shorter breast cancer survival rates. Further experiments revealed that CASC15 promoted the acquisition of stemness properties of breast cancer cells (BSCCs) by competing with endogenous RNA for miR-654-5p, resulting in overexpression of MEF2D in BCSCs. Overall, breast cancer stemness and tumor development are regulated via the CASC15/miR-654-5p/MEF2D axis. Accordingly, this pathway can be explored for breast cancer therapy.

A comprehensive analysis of transcriptome and phenolic compound profiles suggests the role of flavonoids in cotyledon greening in Catharanthus roseus seedling.

During seedling photo-morphogenesis, cotyledon greening is a vital developmental process and a moment of responding to light stress. An increasing number of reports suggest the function of natural antioxidant protection of phenolic compounds in plant growth and development processes. Due to the antioxidant functions, flavonoids allow plants to respond to abiotic or biotic stresses. As one of the plants rich in secondary metabolites, Catharanthus roseus has drawn great academic interest due to its richness of diverse secondary metabolites with medicinal values. To assess the distribution and function of phenolic compounds during cotyledon greening, combined phenolic profiling and transcriptome were applied in C. roseus seedling through ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometer (UPLC-Q-TOF/MS) and high throughput RNA sequencing, respectively. Results herein showed that light-exposed greening cotyledon accumulated large amounts of C6C3C6-type flavonoids, suggesting the function in repressing reactive oxygen species (ROS) generation to improve light adaptation and seedling survival. Moreover, synergistic up-regulation of relevant genes involved in flavonoids pathway, including PAL, C4H, CHS, FLS, and F3'H, was monitored in response to light. Several crucial candidate transcription factors including bHLH, MYB, and B-box families were likely to function, and thereinto, CrHY5 (CRO_T122304) and CRO_T137938 revealed a prompt response to light, supposing to induce flavonoids accumulation by targeting CHS and FLS. Therefore, this study provided new insight into the potential regulation and underlying roles of flavonoids to improve light acclimation during cotyledon greening.

Development and validation of nomograms for predicting axillary non-SLN metastases in breast cancer patients with 1-2 positive sentinel lymph node macro-metastases: a retrospective analysis of two independent cohorts.

BACKGROUND: It is reported that appropriately 50% of early breast cancer patients with 1-2 positive sentinel lymph node (SLN) micro-metastases could not benefit from axillary lymph node dissection (ALND) or breast-conserving surgery with whole breast irradiation. However, whether patients with 1-2 positive SLN macro-metastases could benefit from ALND remains unknown. The aim of our study was to develop and validate nomograms for assessing axillary non-SLN metastases in patients with 1-2 positive SLN macro-metastases, using their pathological features alone or in combination with STMs. METHODS: We retrospectively reviewed pathological features and STMs of 1150 early breast cancer patients from two independent cohorts. Best subset regression was used for feature selection and signature building. The risk score of axillary non-SLN metastases was calculated for each patient as a linear combination of selected predictors that were weighted by their respective coefficients. RESULTS: The pathology-based nomogram possessed a strong discrimination ability for axillary non-SLN metastases, with an area under the receiver operating characteristic (ROC) curve (AUC) of 0.727 (95% CI: 0.682-0.771) in the primary cohort and 0.722 (95% CI: 0.653-0.792) in the validation cohort. The addition of CA 15-3 and CEA can significantly improve the performance of pathology-based nomogram in the primary cohort (AUC: 0.773 (0.732-0.815) vs. 0.727 (0.682-0.771), P < 0.001) and validation cohort (AUC: (0.777 (0.713-0.840) vs. 0.722 (0.653-0.792), P < 0.001). Decision curve analysis demonstrated that the nomograms were clinically useful. CONCLUSION: The nomograms based on pathological features can be used to identify axillary non-SLN metastases in breast cancer patients with 1-2 positive SLN. In addition, the combination of STMs and pathological features can identify patients with patients with axillary non-SLN metastases more accurately than pathological characteristics alone.

CircRNA-UBE2G1 regulates LPS-induced osteoarthritis through miR-373/HIF-1a axis.

Osteoarthritis (OA) is a very common chronic and degenerative joint disease characterized by persistent destruction of articular cartilage. Recently, increasing evidence showed that circular RNAs (circRNAs) play critical roles in OA progression. However, the functions of circRNAs in OA and their underlying mechanisms of action remain unclear. In the present study, the expression levels of circRNA-UBE2G1 and HIF-1a were significantly increased in OA tissues, whereas miR­373 expression was downregulated. Function assays showed that circRNA-UBE2G1 inhibition reduced the effects of LPS on C28/I2 cells viability and apoptosis. In terms of mechanism, we revealed that circRNA-UBE2G1 binds to miR­373 as competing endogenous RNAs (ceRNAs). HIF-1a might act as a target of miR­373. Moreover, miR­373 suppression or HIF-1a overexpression restored the effects of circRNA-UBE2G1 downregulation on LPS-induced chondrocytes injury. Collectively, our data suggest that circRNA-UBE2G1 facilitates the progression in the LPS-induced OA cell model via regulating the miR­373/HIF-1a axis. ABBREVIATIONS: OA: Osteoarthritis; Circular RNAs; miRNAs: MicroRNAs; Mut: Mutant; WT: Wild type; UTR: Untranslated region.

Comparative Study of Growth, Cadmium Accumulation and Tolerance of Three Chickpea (Cicer arietinum L.) Cultivars.

Trace metals (TM) contamination is a severe problem in the environment and produced an adverse effect on the productivity of crops. Cadmium (Cd) is a TM ranked seven among the top 20 pollutants due to its high toxicity and solubility in water, taken up by the plants and affects their growth and metabolism. In this study, we evaluated the growth, Cd accumulation and tolerance capacities of three chickpea (Cicer arietinum L.) cultivars (NC234 (NC2), ICCV89310 (IC8) and ICCV89323-B (IC8-B)), subjected to two Cd concentrations (25 and 50 µM) in hydroponic culture. The toxicity of Cd reduced the plant height and fresh and dry biomass in all cultivars. The maximum reduction was observed at 50 µM of Cd. Compared with IC8-B, cultivars IC8 and NC2 exhibited better performance with high growth, biomass, root to shoot (R/S) ratio and water content under high Cd stress. To measure the accumulation of Cd in root and shoot, an inductively coupled plasma optical emission spectrometer (ICP-OES) was used. IC8 and NC2 had comparatively high Cd tolerance and accumulation ability (> 100 µg g-1 dry weight), with IC8 being more tolerant and accumulated higher Cd in shoot than NC2, while cultivar IC8-B was sensitive. Root accumulated more Cd than shoot in a dose-dependent manner. The bioconcentration factors (BCF) and bioaccumulation coefficients (BAC) were far higher than one (> 1) and increased with an increase in Cd concentrations, while the translocation factor (TF) was less than one (< 1), suggesting that all the three cultivars were unable to transfer Cd from the root to the shoot efficiently. Our results indicated that IC8 and NC2 proved to be resistant, while IC8-B showed sensitivity when exposed to high Cd stress (50 µM).

MiR-140-5p inhibits cell proliferation and metastasis by regulating MUC1 via BCL2A1/MAPK pathway in triple negative breast cancer.

Noncoding RNAs play important roles in the progression of malignant tumors, including triple negative breast cancer (TNBC). Accumulating evidence supported the involvement of the oncogenic MUC1 in tumor metastasis. Our study aimed to explore the roles of miR-140-5p and MUC1 in TNBC and identify the potential underlying mechanisms. In the present study, we found that miR-140-5p expression was significantly decreased in TNBC tissues and associated with advanced clinical features and poor prognosis. MiR-140-5p overexpression suppressed TNBC cells proliferation, invasion ability in vitro and reduced tumor growth in vivo. Subsequently, MUC1 was verified to be a direct target of miR-140-5p in TNBC. Furthermore, we revealed that MUC1 could regulate MAPK pathway through regulating BCL2A1 expression in TNBC. Thus, our study indicated that miR-140-5p might regulate MUC1 to suppress TNBC cells proliferation and metastasis by regulating BCL2A1/MAPK pathway, suggesting miR-140-5p could serve as a potential therapeutic target for TNBC.

Herbacetin treatment remitted LPS induced inhibition of osteoblast differentiation through blocking AKT/NF-κB signaling pathway.

Inflammation, a common situation during the process of bone healing, is reported to play a negative role in bone regeneration. Up to date, therapeutic strategies for inflammation triggered inhibition of osteoblast differentiation are still limited. The aim of this study was to explore the potential roles and molecular mechanisms of Herbacetin in the process of osteoblast differentiation under LPS-mediated inflammatory environment. By using MC3T3-E1, C2C12 and primary mouse calvarial osteoblast (PMCO) cells as experimental models, we observed that LPS stimulation suppressed osteoblast differentiation via inhibiting alkaline phosphatase (ALP) activity and the expression of several osteoblastic genes (osterix, runx2 and osteocalcin). However, the negative role of LPS during osteoblast differentiation could be restored by Herbacetin treatment. Mechanistical studies revealed that Herbacetin treatment suppressed AKT activation and in turn blocked NF-κB signaling pathway. Furthermore, reactivating AKT by a selective PTEN inhibitor SF1670 suppressed the effect of Herbacetin. These data suggested that Herbacetin might play a protective role in osteoblast differentiation in MC3T3-E1/C2C12/PMCO cells under LPS stimulation.

Transcriptomics comparison reveals the diversity of ethylene and methyl-jasmonate in roles of TIA metabolism in Catharanthus roseus.

BACKGROUND: The medicinal plant, Catharanthus roseus (C. roseus), accumulates a wide range of terpenoid indole alkaloids (TIAs). Ethylene (ET) and methyl-jasmonate (MeJA) were previously reported as effective elicitors for the production of various valuable secondary metabolites of C. roseus, while a few ET or MeJA induced transcriptomic research is yet reported on this species. In this study, the de-novo transcriptome assembly of C. roseus is performed by using the next-generation sequencing technology. RESULTS: The result shows that phenolic biosynthesis genes respond specifically to ET in leaves, monoterpenoid biosynthesis genes respond specifically to MeJA in roots. By screening the database, 23 ATP-binding cassette (ABC) transporter partial sequences are identified in C. roseus. On this basis, more than 80 key genes that encode key enzymes (namely TIA pathway, transcriptional factor (TF) and candidate ABC transporter) of alkaloid synthesis in TIA biosynthetic pathways are chosen to explore the integrative responses to ET and MeJA at the transcriptional level. Our data indicated that TIA accumulation is strictly regulated by the TF ethylene responsive factor (ERF) and bHLH iridoid synthesis 1 (BIS1). The heatmap, combined with principal component analysis (PCA) of C. roseus, shows that ERF co-expression with ABC2 and ABC8 specific expression in roots affect the root-specific accumulation of vinblastine in C. roseus. On the contrast, BIS1 activities follow a similar pattern of ABC3 and CrTPT2 specific expression in leaves, which affects the leaf-specific accumulation of vindoline in C. roseus. CONCLUSIONS: Results presented above illustrate that ethylene has a stronger effect than MeJA on TIA induction at both transcriptional and metabolite level. Furthermore, meta-analysis reveals that ERF and BIS1 form a positive feedback loop connecting two ABC transporters respectively and are actively involved in TIAs responding to ET and MeJA in C. roseus.

Differential responses to Cd stress induced by exogenous application of Cu, Zn or Ca in the medicinal plant Catharanthus roseus.

Cd(II) is one of the most widespread and toxic heavy metals and seriously threatens plant growth, furthermore negatively affecting human health. For survival from this metal stress, plants always fight with Cd(II) toxicity by themselves or using other external factors. The effects of second metals copper (Cu(II)), zinc (Zn(II)) and calcium (Ca(II)) on the Cd(II)-affected root morphology, Cd(II) translocation and metabolic responses in Catharanthus roseus were investigated under hydroponic conditions. We found that the Cd-stressed plants displayed the browning and rot root symptom, excess H2O2 content, lipid peroxidation and Cd(II) accumulation in plants. However, the supplement with second metals largely alleviated Cd-induced toxicity, including browning and rot roots, oxidative stress and internal Cd(II) accumulation. The amended effects at metabolic and transcriptional levels involved in different second metals share either common or divergent strategies. They commonly repressed Cd uptake and promoted Cd(II) translocation from roots to shoots with divergent mechanisms. High Zn(II) could activate MTs expression in roots, while Cu(II) or Ca(II) did not under Cd(II) stress condition. The presence of Ca(II) under Cd stress condition largely initiated occurrence of lateral roots. We then grouped a metabolic diagram integrating terpenoid indole alkaloid (TIA) accumulation and TIA pathway gene expression to elucidate the metabolic response of C. roseus to Cd(II) alone or combined with second metals. The treatment with 100 Cd(II) alone largely promoted accumulation of vinblastine, vindoline, catharanthine and loganin, whereas depressed or little changed the expression levels of genes detected here, compared to 0 Cd(II) control. In the presence of Cd(II), the supplement with second metals displayed specific effect on different alkaloid. Among them, the metal Ca(II) is especially beneficial for serpentine accumulation, Zn(II) mainly promoted tabersonine production. However, the addition of Cu(II) commonly depressed accumulation of most alkaloids detected here. Generally, we presented different mechanisms by which the second metals used to alleviate Cd (II) toxicity. This plant has potential application in phytoremediation of Cd(II), due to relatively substantial accumulation of biomass, as well as secondary metabolites TIAs used as pharmaceutical materials when facing Cd stress.

Utilization of unmodified gold nanoparticles for label-free detection of mercury (II): Insight into rational design of mercury-specific oligonucleotides.

Colorimetric detection of mercury (II) with the use of DNA oligonucleotides and unmodified gold nanoparticles (AuNPs) as indicators has been extensively studied. This study provides in-depth insights into the rational design of mercury-specific oligonucleotides (MSO) in the biosensing system. The leftover bases of MSO, as a result of the formation of T-Hg2+-T base pairs, can adsorb on the AuNPs and hinder their aggregation at concentrations of salt. This phenomenon was directly verified by the changes in particle sizes characterized by dynamic light scattering for the first time. Based on these findings, we proposed a rational design for the MSO with approximately 20-fold improvement in detection sensitivity. The detection limit of the proposed assay decreased to 15nM with a linear working range from 50nM to 300nM for Hg2+. The cross-reactivity against eight other metal ions was negligible compared with the response to Hg2+. Considering the diverse applications of AuNPs with oligonucleotides, this study can serve as a good reference and provides important implications in sensing and DNA-directed nanoparticle assembly.

Optimizations of packed sorbent and inlet temperature for large volume-direct aqueous injection-gas chromatography to determine high boiling volatile organic compounds in water.

For the expanded application area, fast trace analysis of certain high boiling point (i.e., 150-250 °C) volatile organic compounds (HVOCs) in water, a large volume-direct aqueous injection-gas chromatography (LV-DAI-GC) method was optimized for the following parameters: packed sorbent for sample on-line pretreatment, inlet temperature and detectors configuration. Using the composite packed sorbent self-prepared with lithium chloride and a type of diatomite, the method enabled safe injection of an approximately 50-100 µL sample at an inlet temperature of 150 °C in the splitless mode and separated HVOCs from water matrix in 2 min. Coupled with a flame ionization detector (FID), an electron capture detector (ECD) and a flame photometric detector (FPD), the method could simultaneously quantify 27 HVOCs that belong to seven subclasses (i.e., halogenated aliphatic hydrocarbons, chlorobenzenes, nitrobenzenes, anilines, phenols, polycyclic aromatic hydrocarbons and organic sulfides) in 26 min. Injecting a 50 µL sample without any enrichment step, such as cryotrap focusing, the limits of quantification (LOQs) for the 27 HVOCs was 0.01-3 µg/L. Replicate analyses of the 27 HVOCs spiked source and river water samples exhibited good precision (relative standard deviations ≤ 11.3%) and accuracy (relative errors ≤ 17.6%). The optimized LV-DAI-GC was robust and applicable for fast determination and automated continuous monitoring of HVOCs in surface water.

[Realtime analysis of volatile organic compounds in source water by membrane inlet/time-of-flight mass spectrometry].

To establish an early-warning system for source water pollution accident, a membrane inlet/time-of-flight mass spectrometry technology was applied to a series of pollution scenarios as an online monitoring method for typical volatile organic compounds such as BTEX (benzene, toluene, ethylbenzene and xylene), substituted benzenes, and halogenated aliphatic hydrocarbons. It was shown that this technology can adequately meet the requirements of realtime analysis with short response time to the target organic pollutants (30-70 s for BTEX and 30 s for halogenated aliphatic hydrocarbons) in a linear detecting range of 3-4 magnitudes; the detection limits of BTEX and chlorobenzene were less than 10 microg x L(-1). The results of 52 simulated water pollution accidents in a 30-days' continuous monitoring indicated that the monitoring system was stable with the relative standard deviation less than 5%; the accuracy was acceptable and could be reduced to within 10% by periodical calibrations. Membrane inlet/time-of-flight mass spectrometry technology was proven to be available for the remote monitoring and early-warning of source water pollution accident.