Bladder Cancer Invasion Research Articles

Fine particulate matter induces METTL3-mediated m6A modification of BIRC5 mRNA in bladder cancer

Long-term exposure to fine particulate matter (PM2.5) is reportedly related to a variety of cancers including bladder cancer. However, little is known about the biological mechanism underlying this association. In the present study, PM2.5 exposure was significantly associated with increased levels of m6A modification in bladder cancer patients and bladder cells. METTL3 expression was aberrantly upregulated after PM2.5 exposure, and METTL3 was involved in PM2.5-induced m6A methylation. Higher METTL3 expression was observed in bladder cancer tissues and METTL3 knockdown dramatically inhibited bladder cancer cell proliferation, colony formation, migration and invasion, inducing apoptosis and disrupting the cell cycle. Mechanistically, PM2.5 enhanced the expression of METTL3 by inducing the promoter hypomethylation of its promoter and increasing the binding affinity of the transcription factor HIF1A. BIRC5 was identified as the target of METTL3 through m6A sequencing (m6A-Seq) and KEGG analysis. The methylated BIRC5 transcript was subsequently recognized by IGF2BP3, which increased its mRNA stability. In particular, PM2.5 exposure promoted the m6A modification of BIRC5 and its recognition by IGF2BP3. In addition, BIRC5 was involved in bladder cancer proliferation and metastasis, as well as VEGFA-regulated angiogenesis. This comprehensive study revealed that PM2.5 exposure exerts epigenetic regulatory effects on bladder cancer via the HIF1A/METTL3/IGF2BP3/BIRC5/VEGFA network.

Retraction: MicroRNA-137 Upregulation Increases Bladder Cancer Cell Proliferation and Invasion by Targeting PAQR3.

Retraction: MicroRNA-137 Upregulation Increases Bladder Cancer Cell Proliferation and Invasion by Targeting PAQR3.

Novel lncRNA LINC01614 Facilitates Bladder Cancer Proliferation, Migration and Invasion Through the miR-217/RUNX2/Wnt/β-Catenin Axis [Retraction

[This retracts the article DOI: 10.2147/CMAR.S330019.].

MAP9 Exhibits Protumor Activities and Immune Escape toward Bladder Cancer by Mediating TGF-β1 Pathway.

To investigate more potential targets for the treatment of human bladder cancer, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and high-content screening (HCS) analysis were performed, and microtubule-associated protein 9 (MAP9), which had the strongest proliferation inhibition from 809 downregulated genes, has been selected. MAP9 is responsible for bipolar spindle assembly and is involved in the progression of many types of tumors; however, its role in bladder cancer (BC) remains unknown. Expressive levels of MAP9 in BC tissues were determined through immunohistochemistry, and the clinical significance of MAP9 in BC was analyzed. Short hairpin ribonucleic acid- (ShRNA-) MAP9 was used to construct stable MAP9 knockdown BC cell lines. The proliferative abilities of MAP9 were measured through assays in vivo and in vitro, and the migrated and invasive abilities of MAP9 were analyzed via in vitro experiments. Quantitative reverse transcription PCR, western blotting, coimmunoprecipitation (Co-IP), and rescue assays were used to identify downstream targets of MAP9. MAP9 expression increased in the tumor tissues, and its increased level was negatively correlated with prognosis. Further, the loss of MAP9 caused decreased BC cell proliferation via inducing the growth 1/synthesis (G1/S) cell cycle arrest in vitro and slowed tumor growth in vivo. In addition, MAP9 silencing attenuated BC cell migration and invasion. Moreover, we found that the growth 1/synthesis (G1/S) cell cycle-related genes and the epithelial mesenchymal transition (EMT) marker levels decreased after silencing MAP9. Finally, we found that the transforming growth factor beta 1 (TGF-β1) pathway is activated as a mediator for MAP9 to regulate genes related to the G1/S cell cycle and EMT. MAP9 promotes BC progression and immune escape activity through the TGF-β1 pathway and is a potential novel target for therapies of BC.

Identification of the Putative Tumor Suppressor Characteristics of FAM107A via Pan-Cancer Analysis.

Family with sequence similarity 107, member A(FAM107A) was supposed as a tumor suppressor for various types of tumors. However, no pan-cancer analysis of FAM107A is available. Therefore, we conducted a FAM107A-related pan-cancer analysis across thirty-three tumors based on TCGA database to explore the molecular characteristics of FAM107A. The FAM107A expression is reduced in most cancers, and its down-regulated expression was linked to poor overall survival and progression-free survival of tumor patients. Analysis of DNA methylation of the FAM107A gene showed a negative correlation between FAM107A expression and promoter methylation in numerous cancers. Furthermore, FAM107A expression was noted to be involved in myeloid-derived suppressor cell infiltration in multiple cancers. To explore the mechanism of FAM107A in cancers, KEGG, and GO enrichment analysis was performed and the result showed “cell adhesion” and “cAMP signaling pathway” terms as the potential impact of FAM107A on cancers. An experiment in vitro showed FAM107A knockdown promoted the proliferation, migration, and invasion of bladder cancer and renal cancer cells. Our study indicates that FAM107A may be a putative tumor suppressor in bladder cancer and other tumors.

Long Noncoding RNA HAGLROS Promotes the Malignant Progression of Bladder Cancer by Regulating the miR-330-5p/SPRR1B Axis.

Bladder cancer (BC) is the most common genitourinary malignancy worldwide, and its aetiology and pathogenesis remain unclear. Accumulating evidence has shown that HAGLROS is closely related to the occurrence and progression of various cancers. However, the biological functions and underlying mechanisms of HAGLROS in BC remain unknown. In the present study, the expression of HAGLROS in BC was determined by public dataset analysis, transcriptome sequencing analysis, qRT–PCR and ISH assays. Gain- or loss-of-function assays were performed to study the biological roles of HAGLROS in BC cells and nude mouse xenograft model. Bioinformatic analysis, qRT–PCR, western blot, immunohistochemistry, FISH assays, subcellular fractionation assays and luciferase reporter assays were performed to explore the underlying molecular mechanisms of HAGLROS in BC. Here, we found that HAGLROS expression is significantly upregulated in BC tissues and cells, and elevated HAGLROS expression was related to higher pathologic grade and advanced clinical stage, which is significant for BC diagnosis. HAGLROS can enhance the growth and metastasis of BC in vitro and in vivo. Furthermore, miR-330-5p downregulation reversed the BC cells proliferation, migration and invasion inhibited by silencing HAGLROS. SPRR1B silencing restored the malignant phenotypes of BC cells promoted by miR-330--5p inhibitor. Mechanistically, we found that HAGLROS functions as a microRNA sponge to positively regulate SPRR1B expression by sponging miR-330-5p. Together, these results demonstrate that HAGLROS plays an oncogenic role and may serve as a potential biomarker for the diagnosis and treatment of BC.

Context-Based Identification of Muscle Invasion Status in Patients With Bladder Cancer Using Natural Language Processing.

Mortality from bladder cancer (BC) increases exponentially once it invades the muscle, with inherent challenges delineating at the population level. We sought to develop and validate a natural language processing (NLP) model for automatically identifying patients with muscle-invasive bladder cancer (MIBC). All patients with a Current Procedural Terminology code for transurethral resection of bladder tumor (TURBT; n = 76,060) were selected from the Department of Veterans Affairs (VA) database. A sample of 600 patients (with 2,337 full-text notes) who had TURBT and confirmed pathology results were selected for NLP model development and validation. The NLP performance was assessed by calculating the sensitivity, specificity, positive predictive value, negative predictive value, F1 score, and overall accuracy at the individual note and patient levels. In the validation cohort, the NLP model had average overall accuracies of 94% and 96% at the note and patient levels. Specifically, the F1 score and overall accuracy for predicting muscle invasion at the patient level were 0.87% and 96%, respectively. The model classified nonmuscle-invasive bladder cancer (NMIBC) with overall accuracies of 90% and 93% at the note and patient levels. When applying the model to 71,200 patients VA-wide, the model classified 13,642 (19%) as having MIBC and 47,595 (66%) as NMIBC and was able to identify invasion status for 96% of patients with TURBT at the population level. Inherent limitations include a relatively small training set, given the size of the VA population. This NLP model, with high accuracy, may be a practical tool for efficiently identifying BC invasion status and aid in population-based BC research.

TMEM2 expression is downregulated as bladder cancer invades the muscle layer

Cell surface hyaluronidase transmembrane protein 2 (TMEM2), which also serves as a reportedly functions in malignancy of several solid tumors. However, TMEM2 involvement in bladder cancer (BCa) is unknown. Therefore, we investigate potential changes in expression of TMEM2 during BCa invasion and over the course of the epithelial mesenchymal transition (EMT). Immunohistochemical analysis of 127 clinical specimens revealed that TMEM2 expression changed with pathological stage (pT) and infiltration pattern (INF) and was highest in pTa-pT1 of INFa tumors and significantly lower at stages from pTa-pT1 to pT2 or 3 in INFb or INFc. E-cadherin expression was highest in INFa and lowest in INFc, a pattern comparable to TMEM2 expression. TMEM2 protein expression analysis of BCa cell lines showed that muscle-invasive T24 and YTS-1 cells with low TMEM2 expression exhibited EMT phenotypes in vitro, in contrast to high TMEM2-expressing non-muscle invasive RT4 cells. EMT-induced non-muscle invasive RT4 cells also showed significantly decreased plasma membrane expression of TMEM2. Our data suggested TMEM2 expression is higher in non-invasive cancers, whereas invasive cancer cells are less likely to express TMEM2 during muscle-invasion and “partial EMT”.

Detecting the muscle invasiveness of bladder cancer: An application of diffusion kurtosis imaging and tumor contact length

PurposeTo evaluate the diagnostic efficacy of diffusion kurtosis imaging (DKI) parameters and tumor contact length (TCL) among clinical and radiological factors for preoperative prediction of muscle-invasive bladder cancer (MIBC). MethodA total of ninety-seven patients underwent 3.0 T MRI scan with propeller fast spin-echo T2WI, echo planar imaging diffusion-weighted imaging (DWI), and dynamic contrast-enhanced imaging (DCE). Two radiologists independently viewed multiparametric MRI (mpMRI) of each patient, graded the VI-RADS, drew the region of interest (ROI) and measured TCL. Interclass correlation coefficients (ICCs), Kappa statistics, Kolmogorov-Smirnov test, Mann-Whitney U tests, chi-square tests, logistic regression analyses, Hosmer-Lemeshow tests, receiver operating characteristic curve (ROC) analysis, and area under the curve (AUC) were applied. ResultsThe mean Kapp of NMIBC group (0.62 ± 0.01) was significantly lower than that of MIBC group (0.79 ± 0.08). The mean TCL of MIBC group (4.66 ± 1.89) was significantly larger than TCL of NMIBC group (1.88 ± 1.50) (all p < 0.01). At the corresponding cut-off, AUC of TCL, Kapp, VI-RADS and the combination of Kapp and TCL were 0.87, 0.92, 0.90, and 0.95, respectively. TCL and Kapp were risk factors of BC muscle invasion at both univariate and multivariate analysis. ConclusionsKapp performed better than conventional DWI in predicting MIBC. Kapp and TCL were independent risk factors of MIBC and could complement VI-RADS for predicting muscle invasion. The combination of Kapp and TCL had the largest AUC and highest accuracy among all parameters.

Evaluating adipose-derived stem cell exosomes as miRNA drug delivery systems for the treatment of bladder cancer.

ObjectivesExosomes are essential mediators of intercellular communication as they transport proteins and RNAs between cells. Owing to their tumor‐targeting capacity, immune compatibility, low toxicity, and long half‐life, mesenchymal stem cell‐derived exosomes have great potential for the development of novel antitumor strategies. In this context, the role of exosomes produced by adipose‐derived mesenchymal stem cells (ADSCs) for the treatment of bladder cancer (BC) remains unclear. Here, we investigated the use of ADSCs as a source of therapeutic exosomes, as well as their efficacy in delivering the tumor suppressor miR‐138‐5p in BC.MethodsADSCs stably expressing miR‐138‐5p were established using Lentivirus infection, and ADSC‐derived miR‐138‐5p exosomes (Exo‐miR‐138‐5p) were isolated from the cell culture medium. The effect of Exo‐miR‐138‐5p on BC cell migration, invasion, and proliferation was evaluated in vitro using wound healing, transwell invasion, and proliferation assays. The in vivo effect of Exo‐miR‐138‐5p was investigated using a subcutaneous xenograft mouse model.ResultsExo‐miR‐138‐5p prevented the migration, invasion, and proliferation of BC cells in vitro. Moreover, ADSC‐derived exosomes could penetrate tumor tissues and successfully deliver miR‐138‐5p to suppress the growth of xenograft tumors in vivo.ConclusionsThe present results reveal that ADSC‐derived exosomes are an effective delivery vehicle for small molecule drugs in vivo, and exosome‐delivered miR‐138‐5p is a promising therapeutic agent for BC treatment.

Long non-coding RNA long intergenic non-protein coding RNA 1232 promotes cell proliferation, migration and invasion in bladder cancer via modulating miR-370-5p/PIM3 axis.

Increasing evidences have suggested that long non-coding RNAs are critical regulators in the progression of tumor growth. Long intergenic non-protein coding RNA 1232 (LINC01232) was verified as an oncogene in multiple cancers. Nevertheless, its function in bladder cancer (BC) remains to be uncovered. In the current study, we detected LINC01232 expression utilizing quantitative real-time polymerase chain reaction (RT-qPCR) and discovered that LINC01232 was overexpressed in BC cell lines versus normal cell line. Besides, the effect of LINC01232 on BC cell behaviors was measured by colony formation, Cell Counting Kit-8 (CCK-8), transwell, TdT-mediated dUTP Nick-End Labeling and caspase-3/8 activity assays. Functionally, LINC01232 deficiency suppressed cell proliferation, migration and invasion. Next, miR-370-5p was proved to bind with LINC01232 by RNA pull down, RNA-binding protein immunoprecipitation (RIP) and luciferase reporter assays. Furthermore, PIM3 expression was negatively modulated by miR-370-5p and markedly increased in BC cell lines. Moreover, PIM3 silence repressed proliferation, migration and invasion but triggered apoptosis of BC cells. The rescue assays validated that upregulation of PIM3 recovered the effects of LINC01232 silence on the growth of BC cells. To summarize, our study manifested that LINC01232 accelerates BC progression by targeting miR-370-5p/PIM3 axis. Targeting LINC01232 might offer novel insight into BC treatment.

PRMT5-activated c-Myc promote bladder cancer proliferation and invasion through up-regulating NF-κB pathway

AimPRMT5 and c-Myc were considered as oncogene of bladder cancer. Nevertheless, whether the interaction between of PRMT5 and c-Myc affect bladder cancer progress is unknown. Herein, we explore the above points and discuss deeply its’ potential mechanism. Method5637 and T24 cells were study subjects in vitro. Western blot was used to examined the protein expression. CCK8 and transwell assay were used to analyze proliferation and invasion ability. Additionally, xenograft tumor model was established. Mice imaging experiment, Immunochemistry assay and western blot were carried out. ResultWestern blot result showed successful transfection of PRMT5-siRNA and c-Myc-siRNA. PRMT5-siRNA could inhibit c-Myc expression, and decrease the proliferation and invasion of bladder cells. And c-Myc overexpression could reverse inhibitory action caused by PRMT5 silence. And in vitro studies found low-expression of c-Myc reduced proliferation and invasion of tumor cells and make the NF-κB pathway inactivation. In vivo studies also demonstrated that inhibiting PRMT5 could downregulate c-Myc expression and inhibit the bladder cancer progress, and the potential mechanism was likely to be related to NF-κB signaling pathway. ConclusionIn a word, low-expression of PRMT5 suppressed c-Myc, and thus inhibited proliferation and invasion ability of 5637 and T24 cells through NF-κB pathway.

Near-infrared fluorescence imaging with intraoperative administration of indocyanine green for laparoscopic radical prostatectomy: Is it a useful weapon for pelvic lymph node dissection?

Near-infrared fluorescence imaging with indocyanine green has emerging applications in urologic surgery. This technology is strongly used in robotic surgery for several ablative and reconstructive procedures. On the contrary, it is not used at all in the urological laparoscopic surgery. To date, bilateral pelvic lymph node dissection represents the most accurate and reliable staging procedure for the detection of lymph node invasion in prostate cancer and bladder cancer. However, it is not devoid of complications. In this field, indocyanine green fluorescence-guided sentinel lymph node identification is an emerging technique, as accurate staging of urologic cancer could be enhanced by an intraoperative lymphatic mapping. Our goal was to show a high spatial resolution, real-time intraoperative imaging technique to recognize the main lymphatic drainage networks, avoiding at same time lymphatic vessel damage. Furthermore, the use of such an imaging system represents an absolute novelty in the field of urological laparoscopy.

LncRNA PVT1 contributes to invasion and doxorubicin resistance of bladder cancer cells through promoting MDM2 expression and AURKB-mediated p53 ubiquitination.

In most bladder cancer (BC) patients, cancer cells will eventually develop chemical resistance causing increased mortality. This study aimed to explore the mechanism of lncRNA plasmacytoma variant translocation 1 (PVT1) in regulating doxorubicin (ADM) resistance of BC cells. We observed that PVT1 expression was upregulated in ADM-resistant BC cells compared with ADM-sensitive BC cells. Downregulation of PVT1 suppressed ADM-resistant BC cell proliferation and invasion, promoted apoptosis, and increased sensitivity to ADM, while PVT1 overexpression promoted ADM-sensitive BC cell growth and their resistance to ADM. Further study uncovered that PVT1 could interact with and promote mouse double minute 2 (MDM2) expression, and upregulated MDM2-mediated Aurora kinase B (AURKB). Furthermore, Nutlin-3, an inhibitor of MDM2, could counteract the promotive effects of PVT1 overexpression on ADM resistance of ADM-sensitive BC cell, the expression of multidrug-resistance-related proteins, and the inhibition of p53-mediated tumor suppressor genes. And, overexpression of MDM2 or AURKB reversed the promotive effects of PVT1 silence on the ADM sensitivity of ADM-resistant BC cell, and the inhibitory effect on expression multidrug resistance proteins. Mechanically, AURKB increased MDM2-mediated p53 ubiquitination. Taken together, PVT1 promoted BC cell proliferation and drug resistance via elevating MDM2 expression and AURKB-mediated p53 ubiquitination.

Identification of muscle-invasion status in bladder cancer patients using natural language processing and machine learning.

447 Background: Mortality from bladder cancer (BC) increases exponentially once it invades the muscle. At the population level, accurate delineation of these patients is challenging. Methods: To develop and validate a natural language processing (NLP) model for automatically identifying muscle-invasive BC (MIBC) patients, aiding in population-based BC research. All patients with a CPT code for transurethral resection of bladder tumor (TURBT) (N = 76,060) were selected from the Department of Veterans Affairs (VA) Corporate Data Warehouse database. A sample of 600 patients (with 2,337 full-text notes) who had TURBT and confirmed pathology results were selected for NLP model development (500 patients) and validation (100 patients). Muscle-invasion (yes/no), unknown, or no cancer, were confirmed by detailed chart review of pathology notes. The NLP performance was assessed by calculating the sensitivity, positive predictive value (PPV), and overall accuracy at the individual note and patient levels. Results: In the validation cohort, the NLP model had overall accuracy of 88% and 92% at the note and patient levels. Specifically, PPV and specificity for predicting muscle-invasion on note level were 83% and 70%, respectively. The model classified non-muscle invasive BC (NMIBC) with 98% sensitivity at both the note and patient levels. Although the sensitivity for MIBC was 70% for note-level determination, the sensitivity was 86% when evaluated at the patient level. When applying the model to 71,200 patients VA-wide, the model classified 13,642 (19%) as having MIBC and 47,595 (66%) as NMIBC. The NLP model was able to identify invasion status for 96% TURBT patients at the population level. Inherent limitations include relatively small training set given the size of the VA population. Conclusions: This NLP model for identifying muscle-invasion at the population level had high accuracy. The NLP model may be a practical and accurate tool for efficiently identifying BC invasion status and may potentially aid in population-based BC research in the VA.

Long Noncoding RNA LINC02470 Sponges MicroRNA-143-3p and Enhances SMAD3-Mediated Epithelial-to-Mesenchymal Transition to Promote the Aggressive Properties of Bladder Cancer.

Simple SummaryLong noncoding RNAs (lncRNAs) were proposed as novel tumor prognostic markers, including for predicting bladder cancer progression, and the competing endogenous RNA (ceRNA) hypothesis conceived an accessible entry point to discover potential lncRNA candidates. This study indicated that LINC02470 promotes bladder cancer cell viability, migration, invasion, and in vivo tumorigenicity by sponging miR-143-3p and consequently rescuing SMAD3 translation to activate the TGF-β-induced EMT process. These data demonstrate that the LINC02470–miR-143-3p–SMAD3 ceRNA axis directly regulates the major transcription factor of TGF-β signaling, SMAD3, thereby inducing the EMT process in bladder cancer and enhancing the aggressiveness of bladder cancer cells.Bladder cancer progression and metastasis have become major threats in clinical practice, increasing mortality and therapeutic refractoriness; recently, epigenetic dysregulation of epithelial-to-mesenchymal transition (EMT)-related signaling pathways has been explored. However, research in the fields of long noncoding RNA (lncRNA) and competing endogenous RNA (ceRNA) regulation in bladder cancer progression is just beginning. This study was designed to determine potential EMT-related ceRNA regulation in bladder cancer progression and elucidate the underlying mechanisms that provoke aggressiveness. After screening the intersection of bioinformatic pipelines, LINC02470 was identified as the most upregulated lncRNA during bladder cancer initiation and progression. Both in vitro and in vivo biological effects indicated that LINC02470 promotes bladder cancer cell viability, migration, invasion, and tumorigenicity. On a molecular level, miR-143-3p directly targets and reduces both LINC02470 and SMAD3 RNA expression. Therefore, the LINC02470–miR-143-3p–SMAD3 ceRNA axis rescues SMAD3 translation upon LINC02470 sponging miR-143-3p, and SMAD3 consequently activates the TGF-β-induced EMT process. In conclusion, this is the first study to demonstrate that LINC02470 plays a pivotally regulatory role in the promotion of TGF-β-induced EMT through the miR-143-3p/SMAD3 axis, thereby aggravating bladder cancer progression. Our study warrants further investigation of LINC02470 as an indicatively prognostic marker of bladder cancer.

CT-based radiomics to predict muscle invasion in bladder cancer

This study investigated the feasibility of a computed tomography (CT)-based radiomics prediction model to evaluate muscle invasive status in bladder cancer. Patients who underwent CT urography at two medical centers from October 2014 to May 2020 and had bladder urothelial carcinoma confirmed by postoperative histopathology were retrospectively enrolled. In total, 441 cases were collected and randomized into a training cohort (n= 293), an internal testing cohort (n= 73), and an external testing cohort (n= 75). The images were first filtered, and then, 1218 features were extracted. The best features related to muscle invasiveness of bladder cancer were identified by ANOVA. A prediction model was built by using the logistic regression method. Statistical analysis was performed by plotting the receiver operating characteristic curve. Indicators of the diagnostic performance of the prediction model, including sensitivity, specificity, accuracy, and area under curve (AUC), were evaluated. In the training, internal testing, and external testing cohorts, the prediction model diagnosed muscle-invasive bladder cancer with AUCs of 0.885 (95% confidence interval [95% CI] 0.841-0.929), 0.820 (95% CI 0.698-0.941), and 0.784 (95% CI 0.674-0.893), respectively. In the internal testing cohort, the sensitivity, specificity, and accuracy of the model were 0.667 (95% CI 0.387-0.870), 0.845 (95% CI 0.721-0.922), and 0.782 (95% CI 0.729-0.827), respectively. In the external testing cohort, the sensitivity, specificity, and accuracy of the model were 0.742 (95% CI 0.551-0.873), 0.750 (95% CI 0.594-0.863), and 0.782 (95% CI 0.729-0.827), respectively. CT-based radiomics prediction model can evaluate muscle invasiveness of bladder cancer before surgery with a good diagnostic performance. • CT-based radiomics model can evaluate muscle invasive status in bladder cancer. • The radiomics model shows good diagnostic performance to differentiate muscle-invasive bladder cancer from non-muscle-invasive bladder cancer. • This preoperative CT-based prediction method might complement MR evaluation of bladder cancer and supplement biopsy.

DNA Methylome Changes of Muscle- and Neuronal-Related Processes Precede Bladder Cancer Invasiveness.

Simple SummaryUrinary bladder cancer can be therapeutically controlled until it becomes invasive, thus identifying critical molecular processes preceding and promoting the transition from pre-invasive to invasive tumors is of vital medical importance. Here, we tested epigenomic (DNA methylation) and gene expression profiles in non-invasive and invasive bladder cancers. We found methylation changes in the genes related to muscle and neuronal processes that discriminate between two cancer stages. Our results may open new avenues for early diagnosis of pre-invasive tumors by testing methylation profiles of tumor cells present in patients’ urine or biopsies leading to timely therapeutic measures.Bladder cancer (BC) is the ninth leading cause of cancer death with one of the highest recurrence rates among all cancers. One of the main risks for BC development is exposure to nitrosamines present in tobacco smoke or in other products. Aberrant epigenetic (DNA methylation) changes accompanied by deregulated gene expression are an important element of cancer pathogenesis. Therefore, we aimed to determine DNA methylation signatures and their impacts on gene expression in mice treated with N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN), a carcinogen similar to compounds found in tobacco smoke. Following BBN administration mice developed non-invasive or invasive bladder cancers. Surprisingly, muscle- and neuronal-related pathways emerged as the most affected in those tumors. Hypo- and hypermethylation changes were present within non-invasive BC, across CpGs mapping to the genes involved in muscle- and neuronal-related pathways, however, methylation differences were not sufficient to affect the expression of the majority of associated genes. Conversely, invasive tumors displayed hypermethylation changes that were linked with alterations in gene expression profiles. Together, these findings indicate that bladder cancer progression could be revealed through methylation profiling at the pre-invasive cancer stage that could assist monitoring of cancer patients and guide novel therapeutic approaches.

Long Noncoding RNA HAND2-AS1 Suppresses Cell Proliferation, Migration, and Invasion of Bladder Cancer via miR-17-5p/KLF9 Axis.

Bladder cancer (BC) is the most common type of malignant tumor in the genitourinary system. Through the microarray analysis of clinical samples, long noncoding RNA HAND2-AS1 expression was found to be downregulated in BC tissues. However, the function of HAND2-AS1 on BC and underlying mechanism are unclear. In this study, the correlations of HAND2-AS1 with clinicopathological parameters in BC patients were determined. The gain- and loss-of-function experiments were conducted to examine the role of HAND2-AS1 in malignant behaviors of BC cells in vitro and in vivo. Then, we paid attention to miR-17-5p/KLF9 axis to illustrate the molecular mechanism. Results showed that HAND2-AS1 was downregulated in BC tissues, and its overexpression significantly inhibited cell proliferation, migration, and invasion in vitro, as well as tumor growth in vivo. Knockdown of HAND2-AS1 caused an opposite effect on BC cell malignancies. Furthermore, miR-17-5p was shown to be a direct target of HAND2-AS1, and it reversed the inhibitory effect of HAND2-AS1 on BC malignancies. Also, as a downstream factor of miR-17-5p, KLF9 silencing was demonstrated to mediate the role of miR-17-5p inhibitor in BC cell proliferation and invasion. Thus, it suggests that HAND2-AS1 acts as a suppressor in BC development through miR-17-5p/KLF9 axis.

CircRNA CRIM1 regulates the migration and invasion of bladder cancer by targeting miR182/Foxo3a axis.

To explore the molecular mechanism of circRNA CRIM1 in the regulation of bladder cancer by targeting the miR182/Foxo3a axis. 50 pairs of cancer tissues and para-cancerous tissues of patients with bladder cancer were collected. RT-PCR method was used to detect the expression of CRIM1 and miR-182. The association between circRNA CRIM1 and clinical data was analyzed. qPCR was used to measure the expression of circRNA CRIM1 and miR-182 in bladder cancer cell UMUC3 and endothelial cell line HUVEC. CRIM1 genes and miR-182 in UMUC3 cell lines were overexpressed and silenced, respectively, to investigate their effects on invasion and migration of bladder cancer, and to detect the changes of miR182/Foxo3a expression. The association between circRNA CRIM1 and miR182/Foxo3a was determined by bioinformatics analysis. The results showed that there was a significant association between the expression of circRNA CRIM1 and distal migration. The expression of CRIM1 in adjacent tissues was significantly down-regulated and negatively correlated with distal migration. The overexpression of circRNA CRIM1 reduced migration and invasion processes in bladder cancer cells. After circRNA CRIM1 was overexpressed, the miR-182 was significantly down-regulated. The expression levels of Foxo3a mRNA and proteins were up-regulated after miR-182 silencing of bladder cancer cell line UMUC3. miR-182 silencing inhibited invasion and migration of cancer cells to some extent. In bladder cancer cells and tissues, CRIM1 and Foxo3a were significantly down-regulated, miR-182 was significantly up-regulated. circRNA CRIM1 regulated the migration and invasion of bladder cancer by targeting the miR182/Foxo3a axis.