Promotes Bladder Cancer Cell Proliferation Research Articles

ALKBH5 Inhibited Cell Proliferation and Sensitized Bladder Cancer Cells to Cisplatin by m6A-CK2α-Mediated Glycolysis

N6-methyladenosine (m6A) is the most commonly occurring internal RNA modification to be found in eukaryotic mRNA and serves an important role in various physiological events. AlkB homolog 5 RNA demethylase (ALKBH5), an m6A demethylase, belongs to the AlkB family of dioxygenases and has been shown to specifically demethylate m6A in RNA, which is associated with a variety of tumors. However, its function in bladder cancer remains largely unclear. In the present study, we found that the expression of ALKBH5 was downregulated in bladder cancer tissues and cell lines. Low expression of ALKBH5 was correlated with the worse prognosis of bladder cancer patients. Furthermore, functional assays revealed that knockdown of ALKBH5 promoted bladder cancer cell proliferation, migration, invasion, and decreased cisplatin chemosensitivity in the 5637 and T24 bladder cancer cell lines in vivo and in vitro, whereas ALKBH5 overexpression led to the opposite results. Finally, ALKBH5 inhibited the progression and sensitized bladder cancer cells to cisplatin through a casein kinase 2 (CK2)α-mediated glycolysis pathway in an m6A-dependent manner. Taken together, these findings might provide fresh insights into bladder cancer therapy.

MEST promotes bladder cancer cell proliferation, migration and invasion via STAT3/Twist-1-mediated EMT.

BackgroundMesoderm-specific transcript (MEST) has been demonstrated to be a proto-oncogene or anti-oncogene in various carcinomas. However, the role and mechanism of MEST in bladder cancer (BC) are still unknown. Here we aimed to explore the effect of MEST on malignant biological behaviour in BC and its potential mechanism.MethodsThe expression of MEST in BC tissues and cells was detected by qRT-PCR methods. MEST depletion and overexpression cell lines were established in T24 and 5637 respectively. Then the effects of MEST on cell proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) were investigated. Finally, the STAT3/Twist-1 signaling was verified.ResultsMEST was elevated in BC tissues and cells lines, and its high expression was highly relevant to the clinicopathologic features of patients with BC and to poor prognosis in these patients. MEST depletion impeded cell proliferation, migration and invasion as well as epithelial-mesenchymal transition (EMT), while MEST overexpression promoted malignant biological behaviour in BC. Mechanistically, MEST upregulated p-STAT3 and Twist-1 expression, while treatment with a STAT3 inhibitor clearly attenuated the STAT3 activation and Twist-1 upregulation induced by MEST. Subsequently, rescue assays confirmed that inhibition of STAT3 signalling could remarkably relieve the oncogenic effects of MEST on malignant biological behaviour in BC.ConclusionsOur data confirmed that MEST exerts oncogenic functions in bladder cancer via STAT3/Twist-1 signalling and that MEST may represent a promising target in BC treatment.

Isoflurane promotes epithelial-to-mesenchymal transition and metastasis of bladder cancer cells through HIF-1α-β-catenin/Notch1 pathways

AimsEpithelial-to-mesenchymal transition (EMT) facilitates cell migration and invasion, and contributes to metastasis in bladder cancer. Within the perioperative period, anesthetic such as isoflurane have been found to affect cancer prognosis. In the study, we reported the tumor-promoting effect of isoflurane in bladder cancer. Materials and MethodsHuman bladder cancer cell lines T24 and BIU-87 were exposed to isoflurane at different concentrations. The immunofluorescent staining of Ki67, Annexin V-FITC/PI staining, Transwell invasion assays and wound-healing assays were performed to assess cell proliferation, apoptosis, invasion and migration. Expressions of EMT markers (E-cadherin, N-cadherin and Vimentin) and metastatic markers (Snail-1, Slug-1 and MMP-2/9) were determined by immunoblotting. Orthotopic tumor models and mice given tail vein injection of T24 cells were developed with or without 4-h exposure to 2% isoflurane. Key findingsWe found isoflurane promoted bladder cancer cell proliferation, invasion and migration but reduce apoptosis in a concentration-dependent manner. In addition, isoflurane was shown to increase HIF-1α and its nuclear accumulation in bladder cancer cells. HIF-1α knockdown inhibited bladder cancer cell proliferation and delayed EMT, which was reversed in the presence of 4-h exposure to 2% isoflurane. Likewise, we found isoflurane modulated β-catenin/Notch1 pathways via HIF-1α. In vivo studies showed that isoflurane exposure accelerated formation of orthotopic bladder tumor and promoted hepatic metastases from carcinoma of the bladder. SignificanceTaken together, our study demonstrates that a frequently used anesthetic can exert a protumorigenic effect on bladder cancer. Isoflurane may serve as an important contributory factor to high recurrence following surgery.

Downregulation of TET1 Promotes Bladder Cancer Cell Proliferation and Invasion by Reducing DNA Hydroxymethylation of AJAP1.

Ten-eleven translocation 1 (TET1) is a member of methylcytosine dioxygenase, which catalyzes 5-methylcytosine (5 mC) to 5-hydroxymethylcytosine (5 hmC) to promote the demethylation process. The dysregulated TET1 protein and 5 hmC level were reported to either suppress or promote carcinogenesis in a cancer type-dependent manner. Currently, the role of TET1 in the development of urinary bladder cancer (UBC) and its underlying molecular mechanisms remain unclear. Herein, we found that TET1 expression was downregulated in UBC specimens compared with normal urothelium and was inversely related to tumor stage and grade and overall survival, suggesting its negative association with UBC progression. TET1 silencing in UBC cells increased cell proliferation and invasiveness while the ectopic expression of wild-type TET1-CD, but not its enzymatic inactive mutant, reversed these effects and suppressed tumorigenicity in vivo. In addition, as a direct regulator of TET1 activity, vitamin C treatment increased 5 hmC level and inhibited the anchorage-independent growth and tumorigenicity of UBC cells. Furthermore, we found that TET1 maintained the hypomethylation in the promoter of the AJAP1 gene, which codes for adherens junction-associated protein 1. The downregulation of AJAP1 reversed TET1-CD-induced nuclear translocation of β-catenin, thus inhibiting the expression of its downstream genes. In human UBC specimens, AJAP1 is frequently downregulated and positively associated with TET1. Notably, low expression levels of both TET1 and AJAP1 predict poor prognosis in UBC patients. In conclusion, we found that the frequently downregulated TET1 level reduces the hydroxymethylation of AJAP1 promoter and subsequently activates β-catenin signaling to promote UBC development. The downregulation of both TET1 and AJAP1 might be a promising prognostic biomarker for UBC patients.

MP17-10 N 6 -METHYLADENOSINE MODIFICATION OF CIRCANKS1A ACCELERATES CYTOPLASMIC EXPORT AND STABILIZES RACGAP1 TO PROMOTE BLADDER CANCER CELL PROLIFERATION AND METASTASIS

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology II (MP17)1 Apr 2020MP17-10 N6-METHYLADENOSINE MODIFICATION OF CIRCANKS1A ACCELERATES CYTOPLASMIC EXPORT AND STABILIZES RACGAP1 TO PROMOTE BLADDER CANCER CELL PROLIFERATION AND METASTASIS Qiangqiang Ge*, Yang Xun, Junlin Lu, and Shaogang Wang Qiangqiang Ge*Qiangqiang Ge* More articles by this author , Yang XunYang Xun More articles by this author , Junlin LuJunlin Lu More articles by this author , and Shaogang WangShaogang Wang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000842.010AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Circular RNAs (circRNAs) have been identified to serve as significant regulators in cancer progression. However, the role of circRNA in bladder cancer (BCa) remains largely unknown. Herein, we investigate the expression profiles of circRNA in 20 paired samples of BCa. METHODS: To investigate the the expression profiles of circRNA, we performed high-throughput CircRNA Microarray using 20 pairs of BCa and matched adjacent non-tumor tissue samples. To explore crucial circRNAs that involved in BCa, co-expression network analysis was performed between the top 100 mostly changed circRNAs. The network represented that 15 circRNAs might involve in BCa progression. Analysis of the correlation between the expression level of 15 circRNAs and clinical data was conducted and circANKS1A was chosen. Gain-of-function and loss-of-function experiments were used to analyze the effect of circANKS1A on cell proliferation and metastasis in vitro and in vivo. m6A-containing RNAs were precipitated from methylated RNA immunoprecipitation (MeRIP) assays. The role of m6A methylation of circANKS1A in BCa proliferation and metastasis was investigated. To explore the potential molecular mechanisms of circANKS1A in regulating BCa malignance, RNA pull-down assays and mass spectrometry analysis were performed to screen circANKS1A-interacting proteins. RESULTS: By evaluating the co-expression network of differentially expressed circRNAs, we identify an N6-methyladenosine (m6A) modified circRNA, circANKS1A, upregulated in serum samples and tumor tissues from BCa patients with muscle invasive bladder Cancer (MIBC). What's more, circANKS1A upregulation is correlated with higher TNM stage and larger tumor size in BCa patients, and predicts poorer patient survival. Elevated circANKS1A promotes cell proliferation both in vitro and in vivo. The upregulated expression of circANKS1A facilitates cancer cells migration in vitro and promotes lung metastasis and liver metastasis in PDX metastasis models in vivo. Of note, N6-methyladenosine modification of circANKS1A accelerates export to the cytoplasm. By forming a circANKS1A/FOXM1/RACGAP1 RNA-protein ternary complex in the cytoplasm, circANKS1A enhances the stability of RACGAP1 mRNA to promote BCa cell proliferation and metastasis progression. Clinically, the upregulated expressions of circANKS1A and RACGAP1 are more prevalent in MIBC tissues than in non-muscle invasive bladder Cancer (NMIBC) tissues. CONCLUSIONS: N6-methyladenosine modification of circANKS1A modulates cytoplasmic export, stabilizes RACGAP1 to promote BCa cell proliferation and metastasis, and indicate that circANKS1A could serve as a significant prognostic marker and therapeutic target for BCa. Source of Funding: National Foundation Committee of Natural Sciences of China (grant numbers 81570631 and 81770704) © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e229-e230 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Qiangqiang Ge* More articles by this author Yang Xun More articles by this author Junlin Lu More articles by this author Shaogang Wang More articles by this author Expand All Advertisement PDF downloadLoading ...

MiR-20a acted as a ceRNA of lncRNA PTENPL and promoted bladder cancer cell proliferation and migration by regulating PDCD4.

Bladder cancer is the most frequent tumor of the urinary system. Despite variety of new treatment options, bladder cancer remains a main global medical problem. Our purpose was to explore the potential molecular and therapeutic targets of bladder cancer diagnosis. The qRT-PCR was used to assess the expression of miR-20a in tissues and cell lines. Counting Cell Kit-8 (CCK-8) assay was carried out to evaluate cell proliferation. Cell migration was calculated using the transwell assay. The expression of miR-20a increased and PDCD4 decreased in bladder cancer tissues compared with normal tissues. Overexpression of miR-20a promoted T24 cell proliferation and migration, while miR-20a inhibitor suppressed cell proliferation and migration. MiR-20a targeted PDCD4 to regulate its expression in T24 cells. MiR-20a is inversely related to PDCD4 and PTENPL in bladder cancer tissues. Upregulation of PDCD4 suppressed T24 cell proliferation and migration. The PTENP1/miR-20a/PTEN axis was involved in the progression of bladder cancer. Our study investigated the function of miR-20a in bladder cancer and provided new insights into the treatment of bladder cancer.

The  Long Non-Coding RNA TMPO-AS1 Promotes Bladder Cancer Growth and Progression via OTUB1-Induced E2F1 Deubiquitination

Background: Increasing evidence indicates long non-coding RNAs (lncRNAs) play crucial roles in cancer tumorigenesis and progression. TMPO antisense RNA 1 (TMPO-AS1) has been found to be involved in several cancers by acting as an competing endogenous RNA. However, the potential roles of TMPO-AS1 interacting with proteins in bladder cancer (BC) remain poorly understood. Methods: The lncRNA TMPO-AS1 expression was evaluated by bioinformatics analysis and was validated by qRT-PCR. Loss- and gain-of- function assays were performed to determine the biological functions of TMPO-AS1 in BC carcinogenesis. Chromatin immunoprecipitation, luciferase reporter assays, western blotting, RNA pull-down, RNA immunoprecipitation assays, and fluorescence in situ hybridization were conducted to explore the molecular mechanisms of TMPO-AS1/E2F transcription factor 1 (E2F1) loop. Findings: TMPO-AS1 is upregulated in bladder cancer and is associated with poor prognosis of BC patients. Functional experiments demonstrated that TMPO-AS1 promotes bladder cancer cell proliferation, migration, invasion, and inhibits cell apoptosis in vivo and in vitro. Mechanically, E2F1 is responsible for the TMPO-AS1 upregulation. Overexpression of TMPO-AS1 facilitates the interaction of E2F1 with OTU domain-containing ubiquitin aldehyde binding 1 (OTUB1), leads to E2F1 deubiquitination and stabilization, and finally promotes BC malignant phenotypes. Interpretation: Our study is the first to uncover a novel positive regulatory loop of TMPO-AS1/E2F1 important for the promotion of BC malignant phenotypes. The TMPO-AS1/E2F1 loop should be considered in the quest for new BC therapeutic options. Funding Statement: This work was financially supported by the National Natural Science Foundation of China (81874137), Outstanding Youth Foundation of Hunan Province (2018JJ1047), Hunan Province Science and Technology Talent Promotion Project (2019TJ-Q10), and Independent Exploration and Innovation Project of Central South University(2019zzts825). Declaration of Interests: The authors declare no conflicts of interest relevant to this study. Ethics Approval Statement: This study was approved by the ethics committee of the Third Xiangya Hospital, Central South University, and written informed consent were obtained from each patients.

Cancer-associated fibroblasts-derived exosomes-mediated transfer of LINC00355 regulates bladder cancer cell proliferation and invasion.

The aim of this study was to investigate the regulatory mechanism of cancer-associated fibroblasts (CAFs) exosome in bladder cancer (BC) cell proliferation and invasion. CAFs and normal fibroblasts (NFs) were isolated from tumor tissues and adjacent normal tissues of BC patients, and examined by immunocytochemistry for the expression of fibroblast activation protein alpha (FAP) and α-smooth muscle actin (α-SMA). Exosomes were extracted from CAFs and NFs and observed under a transmission electron microscope, and expression of the exosome markers CD9 and CD63 was confirmed by western blotting. The distribution and intensity of fluorescence were observed by confocal laser microscopy to analyze exosomes uptake by BC cell lines T24 or 5367. BC cell proliferation and invasion were detected by MTT and Transwell assays, respectively. LINC00355 levels in CAFs, NFs, CAFs exosome, NFs exosome, and BC cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results showed that CAFs exosome significantly promoted BC cell proliferation and invasion relative to NFs exosome. LINC00355 expression was significantly elevated in CAFs exosome when compared with that in NFs-exosome. Up-regulated LINC00355 expression was observed both in T24 and 5367 cells co-incubated with CAFs exosome. Exosomes derived from LINC00355 siRNA-transfected CAFs observably repressed BC cell proliferation and invasion when compared with control siRNA-CAFs exosome. In conclusion, CAFs exosome-mediated transfer of LINC00355 regulates BC cell proliferation and invasion. Significance of the study. In this study, our data suggest that the exosomes released from CAFs promote BC cell proliferation and invasion. The mechanism of this effect is, at least in part, related to the increased LINC00355. Regulation of LINC00355 expression in exosomes released from CAFs might be a putative therapeutic strategy against the pathogenesis of BC.

P114 - ADNP-mediated cell cycle promotes bladder cancer cell proliferation via AKT pathway

P114 - ADNP-mediated cell cycle promotes bladder cancer cell proliferation via AKT pathway

MiR-652-3p promotes bladder cancer migration and invasion by targeting KCNN3.

Increasing evidence indicated that microRNAs (miRNAs) are crucial regulators for cancer development. Bladder cancer (BCa) is a major threat to human health. The aim of this study was to analyze the roles of miR-652-3p in BCa, and to explore the associated mechanisms. MiR-652-3p expression in BCa cell lines was explored using Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) method. MiR-652-3p expression level in BCa tissues was explored at StarBase. Cell Counting Kit-8 (CCK-8) assay, wound-healing assay, and transwell invasion assay were conducted to investigate the biological roles of miR-652-3p. The underlying mechanisms of miR-652-3p in NSCLC were investigated using luciferase activity reporter assay and rescue experiments. We showed that miR-652-3p expression level was upregulated in both BCa tissues and cell lines. The knockdown of miR-652-3p significantly inhibited BCa cell proliferation, migration, and invasion in vitro. Moreover, we showed that potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3 (KCNN3) was a functional target for miR-652-3p. Besides, the expression of KCNN3 in BCa tissues was negatively correlated with miR-652-3p. Collectively, these results showed that miR-652-3p could promote BCa cell proliferation, migration, and invasion via directly regulating KCNN3, which may provide a novel therapeutic target for BCa treatment.

Long non-coding RNA XIST promotes cell proliferation and migration through targeting miR-133a in bladder cancer.

The long non-coding RNA (lncRNA) X inactive specific transcript (XIST) has recently been reported to promote the malignant progression of bladder cancer through regulating several microRNAs (miRs), including miR-124, miR-139-5p and miR-200c. However, whether other miRs are also involved in this process has remained to be determined. The present study demonstrated that XIST was significantly upregulated in bladder cancer tissues compared with that in adjacent normal tissues. Furthermore, its expression was reduced in several common bladder cancer cell lines. High expression of XIST was significantly associated with tumour progression and poor prognosis of patients with bladder cancer. An in vitro experiment indicated that knockdown of XIST significantly reduced the proliferation and migration of bladder cancer cells. A luciferase assay suggested that XIST binds to its predicted binding site in miR-133a. In addition, it was identified that miR-133a was significantly downregulated in bladder cancer, and its expression levels were inversely correlated with those of XIST in bladder cancer tissues. Furthermore, loss- and gain-of-function experiments indicated that miR-133a acted as a downstream effector in XIST-mediated bladder cancer cell proliferation and migration. In conclusion, the present study demonstrates that XIST promotes bladder cancer cell proliferation and migration via targeting miR-133a and thus suggests that XIST may be used as a potential therapeutic target for bladder cancer.

Adenylate kinase 4 promotes bladder cancer cell proliferation and invasion.

Bladder cancer is the second most common urological cancer worldwide with low early diagnosis and high mortality. Since the time of diagnosis directly affects survival rate, early detection and precise biomarkers of bladder cancer are very important. Adenylate kinase 4 (AK4) is a key enzyme involved in cellular metabolism and multiple cancer development; however, the potential role of AK4 in bladder tumorigenesis is still unclear. Immunohistochemistry assay was conducted to evaluate the expression level of AK4 in 107 human bladder cancer tissues. Overall survival and recurrence-free survival were used to assess the prognosis of patients. Colony formation and MTT assays [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide] were performed to measure the proliferation capacity of tumor cells. Cell scratch assays and transwell assays were performed to measure the invasion capacity of tumor cells. The expression level of involved genes was measured by reverse transcription-polymerase chain reaction and western blot assays. The animal model was used to examine the effects of indicated protein on tumorigenesis and invasion in vivo. Herein, our study demonstrated that increased AK4 expression in patients with bladder cancer was associated with a poor prognosis. We further found that inhibition of AK4 in bladder cancer cell line T24 and 5637 can obviously inhibit the proliferation of cancer cells. Transwell assay results showed that down-regulated AK4 was related to the decreased metastasis of T24 and 5637 cells. In addition, AK4-shRNA transfected obviously inhibited tumor growth and metastasis in mice compared with the scramble group. Taken together, the results provide strong evidence of the involvement of AK4 in the progression of bladder cancer and suggest that it could have high potential as a therapeutic target of disease.

LncRNA ITGB1 promotes the development of bladder cancer through regulating microRNA-10a expression.

This study aims to investigate the expression level of lncRNA ITGB1 both in bladder cancer (BCa) tissue and cell lines, as well as to evaluate its function and potential mechanism in the progression of BCa. The expressions of lncRNA ITGB1 in 36 BCa tissues samples (and corresponding adjacent normal ones) and cell lines were detected by quantitative real-time polymerase chain reaction (qRT-PCR). After transfection of sh-ITGB1 in BCa cell lines, the effect of ITGB1 on the proliferation of BCa cells was examined by cell counting kit-8 (CCK-8) assay and colony formation assay. Subsequently, qRT-PCR was used to examine microRNA-10a expression in BCa tissues and cells after ITGB1 was silenced. At the same time, the correlation between ITGB1 and microRNA-10a expression was analyzed. Finally, cell recovery experiment was applied for the in-depth study of the interaction between ITGB1 and microRNA-10a and its underlying mechanism. LncRNA ITGB1 was found upregulated in BCa tissues and cell lines. Knockdown of lncRNA ITGB1 remarkably inhibited cell proliferation. The expression levels of ITGB1 and microRNA-10a in BCa tissues were negatively correlated. ITGB1 downregulation was found to be able to enhance microRNA-10a expression, suggesting that microRNA-10a may be a potential target for ITGB1 in BCa. In addition, cell reverse experiment also verified that ITGB1 could regulate the expression of microRNA-10a, and their interaction affected the malignant progression of BCa. LncRNA ITGB1 level is upregulated in BCa tissues and associated with the pathological stage of BCa, which could be used as a new predictor of BCa patients' prognosis. In addition, ITGB1 might promote BCa cell proliferation via regulating microRNA-10a expression.

Kinesin family member C1 accelerates bladder cancer cell proliferation and induces epithelial-mesenchymal transition via Akt/GSK3β signaling.

Kinesin family member C1 (KIFC1) is implicated in the clustering of multiple centrosomes to maintain tumor survival and is thought to be an oncogene in several kinds of cancers. In our experiments, we first performed bioinformatics analysis to investigate the expression levels of KIFC1 in bladder cancer (BC) specimens and normal bladder epitheliums and then, using our samples, verified findings by quantitative real‐time PCR and western blotting assays. All data showed that KIFC1 was significantly upregulated in BC specimens at both the mRNA and protein levels. Immunohistochemical studies in a cohort of 152 paraffin‐embedded BC tissues displayed that upregulated expression of KIFC1 clearly correlated with pT status (P = .014) and recurrent status (P = .002). Kaplan‐Meier survival analysis and log‐rank test indicated that patients with BC with high KIFC1 expression had both shorter cancer‐specific survival (P < .001) and recurrence‐free survival time (P < .001) than those with low KIFC1 expression. Furthermore, ectopic downregulation of KIFC1 weakened BC cell proliferation and migration both in vitro and in vivo, whereas upregulation of KIFC1 enhanced this in vitro. Overexpression of KIFC1 phosphorylated GSK3β and promoted Snail through activating AKT (protein kinase B0) to induce proliferation and epithelial–mesenchymal transition (EMT) and, therefore, substantially promoted BC migration and metastasis. Our study revealed an oncogenic role for KIFC1 to promote BC cell proliferation and EMT via Akt/GSK3β signaling; KIFC1 might be a promising prognostic biomarker as well as a therapeutic target for BC.

LINC00978 promotes bladder cancer cell proliferation, migration and invasion by sponging miR‑4288.

Long noncoding RNAs regulate the occurrence and progression of numerous types of cancer, including bladder cancer (BCa). However, the role of long intergenic non‑protein coding RNA 978 (LINC00978) in BCa remains unknown. In the present study, it was identified that the LINC00978 expression level was significantly increased in BCa tissues compared with adjacent normal tissues. By performing cell proliferation, colony formation, Transwell and Matrigel assays, it was demonstrated that LINC00978 knockdown significantly inhibited the proliferation, migration and invasion of BCa cells. Conversely, overexpression of LINC00978 resulted in opposite effects. Mechanistically, it was demonstrated that LINC00978 served as a competing endogenous RNA to sponge microRNA‑4288 (miR‑4288), and LINC00978 knockdown significantly increased the expression level of miR‑4288 in BCa cells. Furthermore, it was demonstrated that the expression of LINC00978 was inversely correlated with the miR‑4288 expression level in BCa tissues. Furthermore, by performing a rescue assay, it was identified that inhibition of miR‑4288 was able to reverse the effects of LINC00978 knockdown on proliferation, migration and invasion of BCa cells. Collectively, the present study demonstrated that LINC00978 promoted BCa progression by sponging miR‑4288, suggesting that LINC00978 may represent a potential therapeutic target for BCa treatment.

LncRNA CCAT1 promotes bladder cancer cell proliferation, migration and invasion.

ABSTRACTObjective:To study the expression patterns of long noncoding RNA (lncRNA) colon cancer-associated transcript 1 (CCAT1) and the changes in cell proliferation, apoptosis, migration and invasion induced by silencing CCAT1 in bladder cancer cells.Materials and Methods:The expression levels of CCAT1 were determined using realtime quantitative polymerase chain reaction in cancerous tissues and paired normal tissues from 34 patients with bladder cancer. The relationship between clinical characteristics and CCAT1 expression was analyzed. And then we conducted cell experiments. Bladder urothelial carcinoma cell lines T24 and 5637 cells were transfected with CCAT1 small interfering RNA (siRNA) or scramble siRNA. Cell proliferation and apoptosis changes were determined using a Cell Counting Kit-8 (CCK-8) assay and a flow cytometry assay. Migration and invasion changes were measured using a wound healing assay and a trans-well assay. microRNAs (miRNAs) were predicted by Starbase 2.0, and their differential expression levels were studied.Results:CCAT1 was significantly upregulated in bladder cancer (P < 0.05). CCAT1 upregulation was positively related to tumor stage (P = 0.004), tumor grade (P = 0.001) and tumor size (P = 0.042). Cell proliferation, migration and invasion were promoted by abnormally expressed CCAT1. miRNAs miR-181b-5p, miR-152-3p, miR-24-3p, miR-148a-3p and miR-490-3p were potentially related to the aforementioned functions of CCAT1.Conclusion:CCAT1 plays an oncogenic role in urothelial carcinoma of the bladder. In addition, CCAT1 may be a potential therapeutic target in this cancer.

Upregulation of NPL4 promotes bladder cancer cell proliferation by inhibiting DXO destabilization of cyclin D1 mRNA

BackgroundNPL4 is an important cofactor of the valosin-containing protein (VCP)–NPL4–UFD1 complex. The VCP–NPL4–UFD1 has been considered as a ubiquitin proteasome system (UPS) regulator and response to protein degradation. While NPL4 plays important roles in various diseases, little is known about its functions in bladder cancer (BC).MethodsMTT assays and colony forming test were performed to evaluate cell proliferation ability and Western blotting was used to detect protein expression. Cyclin D1 mRNA expression was detected using qRT-PCR, and coimmunoprecipitation (CoIP) was used to detect protein–protein interactions.ResultsNPL4 was upregulated in BC tissue and correlated with poor prognosis. Upregulation of NPL4 promoted cell proliferation while suppression of NPL4 reduced BC cell proliferation. Upregulation of NPL4 led to overexpression of cyclin D1 by enhancing its mRNA stability. Moreover, NPL4 was found to bind directly to DXO and induce its degradation. DXO was downregulated in BC tissue and regulated BC cell proliferation by destabilizing cyclin D1 mRNA. DXO-mediated NPL4 regulated BC cell proliferation by stabilizing cyclin D1 expression.ConclusionsThe NPL4/DXO/cyclin D1 axis exert crucial role in BC cell growth and is associated with prognosis and may represent a potential therapeutic target for BC.

Correction: CLCA4 inhibits bladder cancer cell proliferation, migration, and invasion by suppressing the PI3K/AKT pathway.

[This corrects the article DOI: 10.18632/oncotarget.21724.].

MicroRNA-93 promotes bladder cancer proliferation and invasion by targeting PEDF

ObjectiveMicroRNA-93 (miR-93) is upregulated in the urine of patients with bladder cancer (BC). Here, we investigated the role of miR-93 in BC progression and explored the underlying mechanism. MethodsmiR-93 expression in BC tissues and cells was detected by real time-polymerase chain reaction. The effects of miR-93 and pigment epithelium-derived factor (PEDF) on cell proliferation and invasion were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Transwell assays. The binding of miR-93 to the 3′-untranslated region of PEDF was identified by the luciferase reporter assay. ResultsmiR-93 expression was higher in BC tissues than in normal controls, and its expression was associated with tumor stage and node stage. Inhibition of miR-93 suppressed the proliferation and invasion of BC cells. PEDF was identified as a target of miR-93 and shown to mediate the effect of miR-93 on cell proliferation and invasion. ConclusionsThe present data suggested that miR-93 promoted BC cell proliferation and invasion by targeting PEDF, providing new biomarkers and targets for BC diagnosis and treatment.

Upregulation of Long Noncoding RNA TUG1 Promotes Bladder Cancer Cell Proliferation, Migration, and Invasion by Inhibiting miR-29c.

Bladder cancer (BC) is one of the leading causes of cancer-related deaths in the world. Long noncoding RNA (lncRNA) taurine-upregulated gene 1 (TUG1) plays an important role in the development and progression of numerous cancers, including BC. However, the exact role of TUG1 in modulating BC progression is still poorly known. In this study, we found that TUG1 was upregulated and microRNA-29c (miR-29c) was downregulated in BC tissues and cell lines. Overexpression of TUG1 promoted the cell proliferation of T24 and EJ cells, whereas TUG1 knockdown had the opposite effect. Upregulation of TUG1 obviously facilitated the migration and invasion of T24 and EJ cells. In contrast, TUG1 silencing repressed the migration and invasion of T24 and EJ cells. Furthermore, TUG1 knockdown markedly increased the expression of miR-29c in vitro. On the contrary, overexpression of TUG1 remarkably decreased the expression of miR-29c. Transfection with plasmids containing mutant TUG1 has no effect on the expression of miR-29c. There were direct interactions between miR-29c and the binding sites of TUG1. In addition, the inhibitory effects of small interfering RNA specific for TUG1 on BC cell proliferation, migration, and invasion were reversed by downregulation of miR-29c. Collectively, our study strongly demonstrates that TUG1 promotes BC cell proliferation, migration, and invasion by inhibiting miR-29c, suggesting that lncRNA TUG1 may be a promising target for BC gene therapy.