Co-transcriptional R-loops-mediated epigenetic regulation drives growth retardation and docetaxel chemosensitivity enhancement in advanced prostate cancer.

R-loops are prevalent three-stranded nucleic acid structures, comprising a DNA-RNA hybrid and a displaced single-stranded DNA, that frequently form during transcription and may be attributed to genomic stability and gene expression regulation. It was recently discovered that RNA modification contributes to maintain the stability of R-loops such as N6-methyladenosine (m6A). Yet, m6A-modified R-loops in regulating gene transcription remains poorly understood. Here, we demonstrated that insulin-like growth factor 2 mRNA-binding proteins (IGF2BPs) recognize R-loops in an m6A-dependent way. Consequently, IGF2BPs overexpression leads to increased overall R-loop levels, cell migration inhibition, and cell growth retardation in prostate cancer (PCa) via precluding the binding of DNA methyltransferase 1(DNMT1) to semaphorin 3 F (SEMA3F) promoters. Moreover, the K homology (KH) domains of IGF2BPs are required for their recognition of m6A-containing R-loops and are required for tumor suppressor functions. Overexpression of SEMA3F markedly enhanced docetaxel chemosensitivity in prostate cancer via regulating Hippo pathway. Our findings point to a distinct R-loop resolution pathway mediated by IGF2BPs, emphasizing the functional importance of IGF2BPs as epigenetic R-loop readers in transcriptional genetic regulation and cancer biology.The manuscript summarizes the new role of N6-methyladenosine in epigenetic regulation, we introduce the distinct R-loop resolution mediated by IGF2BP proteins in an m6A-dependent way, which probably lead to the growth retardation and docetaxel chemotherapy resistance in prostate cancer. Moreover, our findings first emphasized the functional importance of IGF2BPs as epigenetic R-loop readers in transcriptional genetic regulation and cancer biology. In addition, our research provides a novel RBM15/IGF2BPs/DNMT1 trans-omics regulation m6A axis, indicating the new crosstalk between RNA m6A methylation and DNA methylation in prostate cancer.

A Class of Activatable NIR-II Photoacoustic Dyes for High-Contrast Bioimaging.

Photoacoustic (PA) imaging is emerging as one of the important non-invasive imaging techniques in biomedical research. Small molecule- second near-infrared window (NIR-II) PA dyes combined with imaging data can provide comprehensive and in-depth in vivo physiological and pathological information. However, the NIR-II PA dyes usually exhibit "always-on" properties due to the lack of a readily optically tunable group, which hinders the further applications in vivo. Herein, a novel class of dyes GX have been designed and synthesized as an activatable NIR-II PA platform, in which the absorption/emission wavelength of GX-5 extends up to 1082/1360 nm. Importantly, the GX dyes have a strong tissue penetration depth and high-resolution for the mouse vasculature structures in NIR-II PA 3D imaging and high signal-to-noise ratio in NIR-II fluorescence (FL) imaging. Furthermore, to demonstrate the applicability of GX dyes, the first NIR-II PA probe GX-5-CO activated by carbon monoxide (CO) was engineered and employed to reveal the enhancement of the CO levels in the hypertensive mice by high-contrast NIR-II PA and FL imaging. We expect that many derivatives of GX dyes will be developed to afford versatile NIR-II PA platforms for designing a wide variety activatable NIR-II PA probes as biomedical tools.

A prospective multi-center randomized comparative trial evaluating outcomes of transrectal ultrasound (TRUS)-guided 12-core systematic biopsy, mpMRI-targeted 12-core biopsy, and artificial intelligence ultrasound of prostate (AIUSP) 6-core targeted biopsy for prostate cancer diagnosis.

BACKGROUND: Artificial intelligence ultrasound of prostate (AIUSP)-targeted biopsy has been used for prostate cancer (PCa) diagnosis. The objective of this prospective multi-center head-to-head clinical randomized comparative trail (RCT) is to compare PCa detection rate in the TRUS-guided 12-core standard systematic biopsy (TRUS-SB) group and cognitive fused mpMRI-guided 12-core biopsy (mpMRI) group against AIUSP group. METHODS: Four hundred patients were randomized to three arms and underwent biopsies by TRUS-SB (n = 133), mpMRI (n = 134), and AIUSP (n = 133) between January 2015 and December 2017. In TRUS-SB group, a standard 12-core systematic biopsy was performed. In mpMRI group, mpMRI-suspicious lesions (PI-RADS 3-5) were targeted by 2-core biopsy followed by a 10-core systematic biopsy. Otherwise, 12-core systematic biopsy was performed. In AIUSP group, a 6-core targeted biopsy was performed. The primary endpoint was PCa detection rate. RESULTS: AIUSP detected the highest rate of PCa (66/133, 49.6%) compared to TRUS-SB (46/133, 34.6%, p = 0.036) and mpMRI (48/134, 35.8%, p = 0.052). Compared to TRUS-SB (35/133, 26.3%) and mpMRI (31/134, 23.1%) groups, clinically significant PCa (csPCa) detection rate was 32.3% (43/133) in AIUSP group. Overall biopsy core positive rate in the TRUS-SB group (11.0%, 176/1598) and in the mpMRI group (12.7%, 204/1608) was significantly lower than that in the AIUSP group (22.7%, 181/798, p < 0.001). CONCLUSIONS: AIUSP detected the highest rate of overall and significant PCa compared to TRUS-SB and mpMRI, and could be used as an alternative to systematic biopsy in the future. REGISTRATION: This trial was registered in ISRCTN (ISRCTN18033113).

Urban expansion patterns and their driving forces analysis: a comparison between Chengdu-Chongqing and Middle Reaches of Yangtze River urban agglomerations.

Urban agglomerations have emerged as the primary drivers of high-quality economic growth in China. While recent studies have examined the urban expansion patterns of individual cities, a comparative study of the urban expansion patterns of urban agglomerations at two different scales is required for a more comprehensive understanding. Thus, in this study, we conduct a two-scale comparative analysis of urban expansion patterns and their driving factors of the two largest urban agglomerations in western and central China, i.e., Chengdu-Chongqing urban agglomeration (CCUA) and the Middle Reaches of Yangtze River urban agglomerations (MRYRUA) at both the urban agglomeration and city levels. We investigate the urban expansion patterns of CCUA and MRYRUA between 2000 and 2020 using various models, including the urban expansion rate, fractal dimension, modified compactness, and gravity-center method. Then we use multiple linear regression analysis and geographically weighted regression (GWR) to explore the magnitude and geographical differentiation of influences for economic, demographic, industrial structure, environmental conditions, and neighborhood factors on urban expansion patterns. Our findings indicate that CCUA experienced significantly faster urban growth compared to MRYRUA. There is an excessive concentration of resources to megacities within the CCUA, whereas there is a lack of sufficient collaboration among the three provinces within the MRYRUA. Additionally, we identify significant differences in the impacts of driving forces of CCUA and MRYRUA, as well as spatial heterogeneity and regional aggregation in the variation of their strength. Our two-scale comparative study of urban expansion patterns will not only provide essential reference points for CCUA and MRYRUA but also serve as valuable insights for other urban agglomerations in China, enabling them to promote sustainable urban management and foster integrated regional development.

Sentiment Analysis on Online Videos by Time-Sync Comments.

Video highlights are welcomed by audiences, and are composed of interesting or meaningful shots, such as funny shots. However, video shots of highlights are currently edited manually by video editors, which is inconvenient and consumes an enormous amount of time. A way to help video editors locate video highlights more efficiently is essential. Since interesting or meaningful highlights in videos usually imply strong sentiments, a sentiment analysis model is proposed to automatically recognize sentiments of video highlights by time-sync comments. As the comments are synchronized with video playback time, the model detects sentiment information in time series of user comments. Moreover, in the model, a sentimental intensity calculation method is designed to compute sentiments of shots quantitatively. The experiments show that our approach improves the F1 score by 12.8% and overlapped number by 8.0% compared with the best existing method in extracting sentiments of highlights and obtaining sentimental intensities, which provides assistance for video editors in editing video highlights efficiently.

In vivo confocal microscopy of sub-basal corneal nerves and corneal densitometry after three kinds of refractive procedures for high myopia.

PURPOSE: To observe corneal nerve fibers and densitometry after small incision lenticule extraction (SMILE), femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK) and laser-assisted subepithelial keratomileusis (LASEK) for high myopia. METHODS: This is a prospective, cross-sectional research study. Patients with high myopia (equivalent spherical lens: -6.00 and -11.00D) who underwent laser corneal refractive surgery were divided into three groups: SMILE, FS-LASIK and LASEK. Scheimpflug imaging of corneal nerves in five areas was observed by confocal microscopy before and 6, 12 months after surgery. Corneal densitometry was measured by Pentacam anterior segment analysis system. RESULTS: Overall, 59 patients were enrolled. The nerve density in the central area did not recover to the preoperative level in three groups until 12 months. The density and length of corneal nerves in central and lower area were better in the SMILE group 6 months postoperatively (p = 0.01), while nerve density did not differ significantly among three groups 12 months postoperatively (p = 0.18). Nerve fibers in central and temporal region were wider in LASEK than that in other two groups at 6- and 12-month follow-up. Corneal densitometry in the central 6 mm diameter was significantly higher in the LASEK group compared with other two groups 6 months postoperatively (p = 0.04). Twelve months postoperatively, corneal densitometry in range of all zone was lower in SMILE than in FS-LASIK and LASEK (p = 0.01, 0.03, 0.04). CONCLUSIONS: Compared with FS-LASIK and LASEK, SMILE-treated eyes with high myopia had certain advantages in nerve density, length and nerve connection way and had better corneal transparency after operation.

Research hotspots of prostate cancer at the international conferences on urology in 2022. / 2022å¹´æ³Œå°¿å¤–ç§‘å›½é™ ä¼šè®®å‰åˆ—è ºç™Œç›¸å ³ç ”ç©¶çƒ­ç‚¹.

The American Urological Association (AUA), European Association of Urology (EUA) and International Urological Society (SIU) annual meetings were held in 2022. Studies on prostate cancer reported in the meetings mainly focus on the advances of diagnostic biomarkers (such as α-2, 3-1inked sialylation of terminal N-glycan on free PSA density, SelectMDx) and imaging techniques [such as multiparametric magnetic resonance imaging, prostate specific membrane antigen(PSMA)-PET/CT], the new method for prostate biopsy, the new treatments of prostate cancer including [177Lu] Ludotadipep and DROP-IN PSMA probe, and the prognosis assessment of prostate cancer (such as AR-V7). This article provides an overview on the research hotspots of three international academic meetings.

A Biocompatible Probe for the Detection of Neutrophil Elastase Free from the Interference of Structural Changes and Its Application to Ratiometric Photoacoustic Imaging In Vivo.

Neutrophil elastase (NE) plays a key role in chronic inflammation and acute responses to infection and injury. Effective disease interventions thus call for precise identification of NE to aid the clinical treatment of such diseases. However, the detection process suffers from the interference of structural changes of NE. Herein, we introduce a molecular probe with high biocompatibility to overcome the interference, which was achieved by combining theoretical calculations and experimental studies, that permits highly specific and sensitive detection of NE in cells and in vivo. The upregulated NE accumulation was specifically measured in inflammation by ratiometric photoacoustic and near-infrared fluorescence imaging, providing a new method for developing more specific fluorogenic probes for other enzymes.

Highly efficient enrichment of intact phosphoproteins by a cadmium ion-based co-precipitation strategy.

Selective separation and enrichment of phosphoproteins are essential for understanding their important functions in almost all cellular processes. Here, taking advantage of the feature that cadmium ion (Cd2+ ) has an overwhelming preference for phosphates, we developed a robust and simple Cd2+ co-precipitation strategy for the selective isolation of intact phosphoproteins. After evaluating the feasibility of Cd2+ in phosphoprotein precipitation, we compared the washing protocols for the removal of non-specific binding proteins and then used the best-performing protocol for the isolation of phosphoproteins from different complex samples. It was found that phosphoproteins can be specifically enriched from artificial protein mixtures containing α-casein, ß-casein, and bovine serum albumin or plasma, in which bovine serum albumin or plasma were served as interferences with very high molar ratios. Applying this method to enrich phosphoproteins from complex cell lysates, a high specificity was confirmed by western blotting analysis with a phosphoprotein-specific kit. Finally, we successfully applied this method to the purification of caseins from drinking milk, highlighting its potential application in the studies where purified phosphoproteins were required. In a word, this Cd2+ co-precipitation method enables universal and effective capture, enrichment, and detection of intact phosphoproteins, making it a powerful tool for the comprehensive analysis of the phosphoproteome.

Small molecule based fluorescent chemosensors for imaging the microenvironment within specific cellular regions.

The microenvironment (local environment), including viscosity, temperature, polarity, hypoxia, and acidic-basic status (pH), plays indispensable roles in cellular processes. Significantly, organelles require an appropriate microenvironment to perform their specific physiological functions, and disruption of the microenvironmental homeostasis could lead to malfunctions of organelles, resulting in disorder and disease development. Consequently, monitoring the microenvironment within specific organelles is vital to understand organelle-related physiopathology. Over the past few years, many fluorescent probes have been developed to help reveal variations in the microenvironment within specific cellular regions. Given that a comprehensive understanding of the microenvironment in a particular cellular region is of great significance for further exploration of life events, a thorough summary of this topic is urgently required. However, there has not been a comprehensive and critical review published recently on small-molecule fluorescent chemosensors for the cellular microenvironment. With this review, we summarize the recent progress since 2015 towards small-molecule based fluorescent probes for imaging the microenvironment within specific cellular regions, including the mitochondria, lysosomes, lipid drops, endoplasmic reticulum, golgi, nucleus, cytoplasmic matrix and cell membrane. Further classifications at the suborganelle level, according to detection of microenvironmental factors by probes, including polarity, viscosity, temperature, pH and hypoxia, are presented. Notably, in each category, design principles, chemical synthesis, recognition mechanism, fluorescent signals, and bio-imaging applications are summarized and compared. In addition, the limitations of the current microenvironment-sensitive probes are analyzed and the prospects for future developments are outlined. In a nutshell, this review comprehensively summarizes and highlights recent progress towards small molecule based fluorescent probes for sensing and imaging the microenvironment within specific cellular regions since 2015. We anticipate that this summary will facilitate a deeper understanding of the topic and encourage research directed towards the development of probes for the detection of cellular microenvironments.

Graph-Based Feature Weight Optimisation and Classification of Continuous Seismic Sensor Array Recordings.

Slope instabilities caused by heavy rainfall, man-made activity or earthquakes can be characterised by seismic events. To minimise mortality and infrastructure damage, a good understanding of seismic signal properties characterising slope failures is therefore crucial to classify seismic events recorded from continuous recordings effectively. However, there are limited contributions towards understanding the importance of feature selection for the classification of seismic signals from continuous noisy recordings from multiple channels/sensors. This paper first proposes a novel multi-channel event-detection scheme based on Neyman-Pearson lemma and Multi-channel Coherency Migration (MCM) on the stacked signal across multi-channels. Furthermore, this paper adapts graph-based feature weight optimisation as feature selection, exploiting the signal's physical characteristics, to improve signal classification. Specifically, we alternatively optimise the feature weight and classification label with graph smoothness and semidefinite programming (SDP). Experimental results show that with expert interpretation, compared with the conventional short-time average/long-time average (STA/LTA) detection approach, our detection method identified 614 more seismic events in five days. Furthermore, feature selection, especially via graph-based feature weight optimisation, provides more focused feature sets with less than half of the original number of features, at the same time enhancing the classification performance; for example, with feature selection, the Graph Laplacian Regularisation classifier (GLR) raised the rockfall and slide quake sensitivities to 92% and 88% from 89% and 85%, respectively.

Inter- and Intra-Modal Contrastive Hybrid Learning Framework for Multimodal Abstractive Summarization.

Internet users are benefiting from technologies of abstractive summarization enabling them to view articles on the internet by reading article summaries only instead of an entire article. However, there are disadvantages to technologies for analyzing articles with texts and images due to the semantic gap between vision and language. These technologies focus more on aggregating features and neglect the heterogeneity of each modality. At the same time, the lack of consideration of intrinsic data properties within each modality and semantic information from cross-modal correlations result in the poor quality of learned representations. Therefore, we propose a novel Inter- and Intra-modal Contrastive Hybrid learning framework which learns to automatically align the multimodal information and maintains the semantic consistency of input/output flows. Moreover, ITCH can be taken as a component to make the model suitable for both supervised and unsupervised learning approaches. Experiments on two public datasets, MMS and MSMO, show that the ITCH performances are better than the current baselines.

A Dual-Stage Attention Model for Tool Wear Prediction in Dry Milling Operation.

The intelligent monitoring of tool wear status and wear prediction are important factors affecting the intelligent development of the modern machinery industry. Many scholars have used deep learning methods to achieve certain results in tool wear prediction. However, due to the instability and variability of the signal data, some neural network models may have gradient decay between layers. Most methods mainly focus on feature selection of the input data but ignore the influence degree of different features to tool wear. In order to solve these problems, this paper proposes a dual-stage attention model for tool wear prediction. A CNN-BiGRU-attention network model is designed, which introduces the self-attention to extract deep features and embody more important features. The IndyLSTM is used to construct a stable network to solve the gradient decay problem between layers. Moreover, the attention mechanism is added to the network to obtain the important information of output sequence, which can improve the accuracy of the prediction. Experimental study is carried out for tool wear prediction in a dry milling operation to demonstrate the viability of this method. Through the experimental comparison and analysis with regression prediction evaluation indexes, it proves the proposed method can effectively characterize the degree of tool wear, reduce the prediction errors, and achieve good prediction results.

Upregulation of ARNTL2 is associated with poor survival and immune infiltration in clear cell renal cell carcinoma.

BACKGROUND: Aryl hydrocarbon receptor nuclear translocator like 2 (ARNTL2) is a member of the PAS superfamily. Previous studies explored the carcinogenic roles of transcription factor ARNTL2 in human malignancies. However, its roles in ccRCC have not been elucidated. This study sought to explore the roles of ARNTL2 in ccRCC and determine its correlations with tumor immunity. METHODS: The expression of ARNTL2 was analyzed using the GEO, TCGA and GTEx database, and verified in ccRCC tissue samples and cell lines by qRT-PCR and western blot analysis. Kaplan-Meier survival curve analysis, Cox regression analysis (including univariate and multivariate analysis) was utilized to evaluate the prognostic values of ARNTL2. Potential biological mechanisms of ARNTL2 were explored using GSEA method. Colony formation and wound healing assays were conducted to explore the oncogenic role of ARNTL2 in ccRCC. ssGSEA and xCell algorithm were used to explore the correlation between ARNTL2 expression and tumor immune microenvironment (TIME). RESULTS: ARNTL2 was significantly upregulated in ccRCC tissues and cell lines compared to normal kidney tissues and cell line. Enhanced expression of ARNTL2 was strongly linked to advanced clinical stage and unfavorable overall survival in ccRCC. ARNTL2 was determined as an independent prognostic marker through cox regression analysis. A prognostic nomogram was constructed to predict 1-, 3- and 5-year overall survival of ccRCC patients by integrating ARNTL2 expression with other clinicopathologic variables. GSEA analysis showed that focal adhesion, T cell receptor, cell cycle, and JAK-STAT signaling pathway were significantly enriched in high ARNTL2 samples. Silencing of ARNTL2 suppressed the colony formation ability and wound healing efficacy of ccRCC cell lines. xCell analysis showed that high expression level of ARNTL2 exhibited an immune infiltration status similar to CD8 + inflamed ccRCC subtype, which was characterized by high infiltration level of CD8 + T cell and high expression level of the immune escape biomarkers such as PD-L1, PD-L2, PD1 and CTLA4. CONCLUSION: ARNTL2 is an independent adverse predictor of ccRCC patient survival. High expression level of ARNTL2 is associated with immune infiltration, and may be a novel therapeutic target in ccRCC.

Investigation on the influence of additives on the oriented dissolution of calcite.

Similar to the crystal growth process, additives have a strong influence on the dissolution process of crystals. Studies on the dissolution process may shed light on understanding the biomineralization and bioinspired crystallization process. The influence of different kinds of additives including surfactants and polymers on the dissolution process of calcite {104} planes was investigated in detail in this work. The additives can be classified into three kinds according to their influence on the dissolution process of calcite under different concentration windows. The additives show three different kinds of dissolution behaviors with the increase of additive concentrations according to the tomographic variation of the calcite surface after the dissolution process. There are four dissolution modes of calcite while changing the additive concentrations in the solution. Rhombohedral etch pits with [4[combining macron]41] and [481[combining macron]] step edges are formed on the calcite {104} planes after the dissolution process at low additive concentrations (mode I). Calcite micropyramids begin to appear on the calcite surface and the densities of micropyramids increase with the increase of the additive concentrations until they cover the entire calcite surface after the dissolution process at medium additive concentrations (mode II). Instead of micropyramids, large pyramids with [481[combining macron]] and [4[combining macron]41] step edges and a size of about 50 µm form after the dissolution process at high additive concentrations (modes III and IV). We propose that the different anisotropic dissolution behaviors of calcite are strongly related to the concentrations and the adsorption features of the additives on the calcite surface. The additives may act as inhibitors of calcite dissolution, possibly through adsorption on calcite surfaces without preferred adsorption, or adsorption at specific kink sites or step edges. The influence of additives on the oriented dissolution of calcite is generally related to the adsorption density and homogeneity of additives on the calcite substrates.

Artisan versus Artiflex phakic intraocular lens implantation in the treatment of moderate to high myopia: meta-analysis.

BACKGROUND: To compare the postoperative safety, efficacy, predictability, visual quality and biomechanics after implantation of Artisan vs. Artiflex phakic intraocular lenses (PIOLs). METHODS: Pubmed, Embase, Cochrane Library were conducted up from January 2000 to February 2020. Comparative clinical studies reporting in accordance with the eligibility criteria were included in this meta-analysis. The pooled weighted mean differences (WMDs) and odds ratios (ORs) with corresponding 95% confidence intervals were calculated. RESULTS: Comparative trials with myopia patients were selected in this review. The pooled WMD and OR estimates statistical significance in terms of postoperative best corrected visual acuity (BCVA), efficacy, postoperative spherical equivalence (SE), predictability, contrast sensitivity and mean intraocular higher-order aberrations (HOA) (mm) for a 6-mm pupil, manifesting that Artiflex PIOL showed evident beneficial effect for correcting myopia compared to Artisan PIOL. There was no significant difference in the incidence of complications between the two groups. CONCLUSION: Both of two techniques were safe and effective for myopia and compared to Artisan PIOL, Artiflex PIOL had significant improvement in efficacy, predictability, contrast sensitivityand HOA, except safety and complications in the treatment of moderate to high myopia.

Understanding the Effects of China's Agro-Environmental Policies on Rural Households' Labor and Land Allocation with a Spatially Explicit Agent-Based Model.

Understanding household labor and land allocation decisions under agro-environmental policies is challenging due to complex human-environment interactions. Here, we develop a spatially explicit agent-based model based on spatial and socioeconomic data to simulate households' land and labor allocation decisions and investigate the impacts of two forest restoration and conservation programs and one agricultural subsidy program in rural China. Simulation outputs reveal that the forest restoration program accelerates labor out-migration and cropland shrink, while the forest conservation program promotes livelihood diversification via increasing non-farm employment. Meanwhile, the agricultural subsidy program keeps labor for cultivation on land parcels with good quality, but appears less effective for preventing marginal croplands from being abandoned. The policy effects on labor allocation substantially differ between rules based on bounded rational and empirical knowledge of defining household decisions, particularly on sending labor out-migrants and engaging in local off-farm jobs. Land use patterns show that the extent to which households pursue economic benefits through shrinking cultivated land is generally greater under bounded rationality than empirical knowledge. Findings demonstrate nonlinear social-ecological impacts of the agro-environmental policies through time, which can deviate from expectations due to complex interplays between households and land. This study also suggests that the spatial agent-based model can represent adaptive decision-making and interactions of human agents and their interactions in dynamic social and physical environments.

Roles of N6 -methyladenosine (m6 A) RNA modifications in urological cancers.

Epigenetics has long been a hot topic in the field of scientific research. The scope of epigenetics usually includes chromatin remodelling, DNA methylation, histone modifications, non-coding RNAs and RNA modifications. In recent years, RNA modifications have emerged as important regulators in a variety of physiological processes and in disease progression, especially in human cancers. Among the various RNA modifications, m6 A is the most common. The function of m6 A modifications is mainly regulated by 3 types of proteins: m6 A methyltransferases (writers), m6 A demethylases (erasers) and m6 A-binding proteins (readers). In this review, we focus on RNA m6 A modification and its relationship with urological cancers, particularly focusing on its roles and potential clinical applications.

METTL3/YTHDF2 m6 A axis promotes tumorigenesis by degrading SETD7 and KLF4 mRNAs in bladder cancer.

N6-Methyladenosine (m6 A) modification, the most prevalent modification of eukaryotic messenger RNA (mRNA), is involved in the progression of various tumours. However, the specific role of m6 A in bladder cancer (BCa) is still poorly understood. In this study, we demonstrated the tumour-promoting function and specific regulatory mechanism of m6 A axis, consisting of the core 'writer' protein METTL3 and the major reader protein YTHDF2. Depletion of METTL3 impaired cancer proliferation and cancer metastasis in vitro and in vivo. Through transcriptome sequencing, m6 A methylated RNA immunoprecipitation (MeRIP) and RIP, we determined that the METTL3/YTHDF2 m6 A axis directly degraded the mRNAs of the tumour suppressors SETD7 and KLF4, contributing to the progression of BCa. In addition, overexpression of SETD7 and KLF4 revealed a phenotype consistent with that induced by depletion of the m6 A axis. Thus, our findings on the METTL3/YTHDF2/SETD7/KLF4 m6 A axis provide the insight into the underlying mechanism of carcinogenesis and highlight potential therapeutic targets for BCa.

YTHDF2 mediates the mRNA degradation of the tumor suppressors to induce AKT phosphorylation in N6-methyladenosine-dependent way in prostate cancer.

BACKGROUND: N6-methyladenosine (m6A) is the most abundant modification in mRNA of humans. Emerging evidence has supported the fact that m6A is comprehensively involved in various diseases especially cancers. As a crucial reader, YTHDF2 usually mediates the degradation of m6A-modified mRNAs in m6A-dependent way. However, the function and mechanisms of m6A especially YTHDF2 in prostate cancer (PCa) still remain elusive. METHODS: To investigate the functions and mechanisms of YTHDF2 in PCa, in vitro, in vivo biofunctional assays and epigenetics experiments were performed. Endogenous expression silencing of YTHDF2 and METTL3 was established with lentivirus-based shRNA technique. Colony formation, flow cytometry and trans-well assays were performed for cell function identifications. Subcutaneous xenografts and metastatic mice models were combined with in vivo imaging system to investigate the phenotypes when knocking down YTHDF2 and METTL3. m6A RNA immunoprecipitation (MeRIP) sequencing, mRNA sequencing, RIP-RT-qPCR and bioinformatics analysis were mainly used to screen and validate the direct common targets of YTHDF2 and METTL3. In addition, TCGA database was also used to analyze the expression pattern of YTHDF2, METTL3 and the common target LHPP in PCa, and their correlation with clinical prognosis. RESULTS: The upregulated YTHDF2 and METTL3 in PCa predicted a worse overall survival rate. Knocking down YTHDF2 or METTL3 markedly inhibited the proliferation and migration of PCa in vivo and in vitro. LHPP and NKX3-1 were identified as the direct targets of both YTHDF2 and METTL3. YTHDF2 directly bound to the m6A modification sites of LHPP and NKX3-1 to mediate the mRNA degradation. Knock-down of YTHDF2 or METTL3 significantly induced the expression of LHPP and NKX3-1 at both mRNA and protein level with inhibited phosphorylated AKT. Overexpression of LHPP and NKX3-1 presented the consistent phenotypes and AKT phosphorylation inhibition with knock-down of YTHDF2 or METTL3. Phosphorylated AKT was consequently confirmed as the downstream of METTL3/YTHDF2/LHPP/NKX3-1 to induce tumor proliferation and migration. CONCLUSION: We propose a novel regulatory mechanism in which YTHDF2 mediates the mRNA degradation of the tumor suppressors LHPP and NKX3-1 in m6A-dependent way to regulate AKT phosphorylation-induced tumor progression in prostate cancer. We hope our findings may provide new concepts of PCa biology.