The original western blot result.

Immunofluorescence staining for CCAT1 in PCa cells

Upregulation CCAT1 in LNCaP could promote UBE2C expression.

DDX5 peptides difference in desthiobiotinylated-CCAT1 sense vs antisense.

The list of miRNA mimics, miRNA inhibitors, primers and shRNA sequence.

CCAT1 expression at different metastatic sites

Pulldown assays showed miR-125a-3p and miR-125b-5p interacted with CCAT1,CCK8 assays showed that miR-125a-3p and miR-125b-5p manifested anti-cancer effect in Du145 cells

The list of miRNAs pulled down by CCAT1 sense and antisense strand in our RNA pulldown assay.

Background The latent involvement of MRPL13 in non-small cell lung cancer (NSCLC) remains unclear. This study aimed to explore the role of MRPL13 in NSCLC. Methods All analyses were performed in R software 4.0, SPSS version 23, and GraphPad Prism 8. The “limma” package was used to identify differentially expressed genes. Univariate and multivariate cox analyses were used to identify prognosis-related genes. A549 and H1299 lung cancer cell lines were selected for phenotypic experiments. Results The high level of MRPL13 was correlated with poor T classification and overall survival. In vitro experiments showed that MRPL13 was highly expressed in NSCLC tissue and cell lines. MRPL13 knockdown inhibited the proliferation of lung cancer A549 and H1299 cell lines, which was further validated by in vivo experiment. Moreover, GSEA analysis suggested that the pathway of MYC target, PI3K/AKT/mTOR/ signaling, oxidative phosphorylation, and G2/M checkpoints may be the potential pathway where MRPL13 was involved. Meanwhile, MRPL13 demonstrated a negative correlation with M1 macrophage, CD8+ T cells, and CD4+ T cells, making it an underlying immunotherapy target of NSCLC. Conclusion MRPL13 may promote the proliferation of NSCLC cells and serve as an independent tumor marker and an emerging therapeutic target.

Although micro RNA (miRNA) expression profiles are widely investigated in renal cell carcinoma (RCC), their potential roles for affecting RCC initiation and progression remain largely unknown. Here, we examined the aberrant expression profiles of miRNAs inhuman metastatic RCC tissues based on Gene Expression Omnibus (GSE37989). We further validated them iRNAs expression data in the largest clinical dataset: The Cancer Genome Atlas (TCGA). And cell adhesion and migration abilities and epithelial me senchymal transition (EMT) related proteins were assessed in both normal and tumor RCC cell lines. We suggest that hsa‐miR‐143 is a potential tumor suppressor in RCC as its down regulation positively correlated with adverse prognosis. Biologically, cell adhesion, migration, and EMT were dramatically inhibited by miR‐143. Mechanistically, we found that miR‐143 targets ABL proto‐oncogene 2 (ABL2), which was also found to be an indicator for poor survival in TCGA database. Our results have important implications in understanding functions of miRNAs in metastatic RCC and will provide a basis for further clinical application.

PURPOSE:To explore the association between MEG3 polymorphisms and the risk of prostate cancer in the Chinese Han population.MATERIALS AND METHODS:Two MEG3 single-nucleotide polymorphisms (SNPs) (rs11627993 C >T rs7158663 A>G) were genotyped in a case-control study in which 165 prostate cancer patients and 200 healthy controls were recruited by a Real-Time Polymerase Chain Reaction (PCR) with the TaqMan assay. The odds ratios (ORs) and 95% confidence intervals (CIs) were used to estimate the strength of association.RESULTS:No statistically significant differences were found in the allele or genotype distributions of the MEG3 rs11627993 C >T and rs7158663 A > G polymorphisms among cases or healthy control subjects (rs11627993: CC vs CA: 95% CI = 0.54-1.95, ORs = 1.03; CC vs AA: 95% CI = 0.67-2.54, ORs = 1.30 ; CC/CA vs AA: 95% CI = 0.81-1.98, ORs = 1.26 , P = .29 ; C vs A: 95% CI = 0.85-1.57, ORs = 1.16, P = .35; rs7158663: AA vs AG: 95% CI = 0.76-5.08, ORs = 1.97, AA vs GG: 95% CI = 0.57-3.29, ORs = 1.37; AA/AG vs GG : 95% CI = 0.56-1.32, ORs = 0.86, P = .49; A vs G: 95% CI = 0.69-1.39, ORs = 0.98, P = .91) Further stratified analysis detected no significant association.CONCLUSION:The MEG3 polymorphisms (rs11627993 C>T and rs7158663 A>G) does not influence the susceptibility to prostate cancer.

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology III (MP81)1 Apr 2019MP81-14 LNCRNA CCAT1 PROMOTES PROSTATE CANCER CELL PROLIFERATION BY INTERACTING WITH DDX5 AND MIR-28-5P Zonghao You*, Chunhui Liu, Can Wang, Bin Xu, Han Guan, and Ming Chen Zonghao You*Zonghao You* More articles by this author , Chunhui LiuChunhui Liu More articles by this author , Can WangCan Wang More articles by this author , Bin XuBin Xu More articles by this author , Han GuanHan Guan More articles by this author , and Ming ChenMing Chen More articles by this author View All Author Informationhttps://doi.org/10.1097/01.JU.0000557433.41195.d2AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVES: Accumulated evidence indicates that CCAT1 functions as an oncogene in the progression of a variety of tumors. However, little is known as to how CCAT1 impacts tumorigenesis in human prostate cancer.Our purpose is to explore the role of CCAT1 in the malignant progression of prostate cancer and the mechanism of the influence. METHODS: Employing qRT-PCR, ISH, and a re-analysis of The Cancer Genome Atlas (TCGA) and The Memorial Sloan Kettering Cancer Center (MSKCC) data, we compared CCAT1 expression between castration-resistant prostate cancer (CRPC) tissues and androgen dependent prostate cancer (ADPC) tissues. Kaplan-Meier method was applied to analysis the prognostic value. The impacts of CCAT1 on biological function were evaluated by in vitro assays and in vivo nude mice model. RNA pulldown, RIP and ChIP-qPCR assays were adopted to explore the regulatory mechanisms of CCAT1 in the progression of CRPC. RESULTS: CCAT1 could promote PCa cell proliferation and accelerate the tumor growth of PCa xenografts. At a molecular level, CCAT1 sponged miR-28-5p to reverse the anti-cancer effect. Additionally, RNA pulldown, RIP and ChIP-qPCR assays revealed CCAT1 acts as a scaffold for DDX5 (P68) and the AR transcriptional complex, thus stimulating CRPC progression. CONCLUSIONS: Our findings indicated CCAT1 acts as one of oncogenic factors in the progression of CRPC by performing different regulatory mechanisms in the nucleus and cytoplasm of cells. Source of Funding: This study was funded by The National Natural Science Foundation of China (No. 81872089, 81370849, 81300472, 81070592, 81202268, 81202034), Natural Science Foundation of Jiangsu Province (BK 20150642), National Science Foundation of Anhui Province (KJ2018A0214). Nanjing, China, People's Republic of; Bengbu, China, People's Republic of; Nanjing, China, People's Republic of© 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 201Issue Supplement 4April 2019Page: e1176-e1177 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Zonghao You* More articles by this author Chunhui Liu More articles by this author Can Wang More articles by this author Bin Xu More articles by this author Han Guan More articles by this author Ming Chen More articles by this author Expand All Advertisement PDF downloadLoading ...

Accumulated evidence indicates that CCAT1 functions as an oncogene in the progression of a variety of tumors. However, little is known as to how CCAT1 impacts tumorigenesis in human prostate cancer. In this study, we found from The Cancer Genome Atlas and Memorial Sloan Kettering Cancer Center database that CCAT1 is highly upregulated in castration-resistant prostate cancer (CRPC) compared with androgen-dependent prostate cancer (ADPC). Higher level of CCAT1 leads to increased mortality in patients with CRPC. In vitro and in vivo studies show that CCAT1 promotes prostate cancer cell proliferation as well as the tumor growth of prostate cancer xenografts. Mechanistically, in cytoplasm, CCAT1 sponges MIR-28-5P to prevent the anticancer effect. In nucleus, CCAT1 acts as a scaffold for DDX5 (P68) and AR transcriptional complex to facilitate the expression of AR-regulated genes, thus stimulating CRPC progression. Our findings suggest that CCAT1 is an oncogenic factor in the progression of CRPC with different regulatory mechanisms in the nucleus and cytoplasm of cells.

Prostate cancer is still considered a significant health care challenge worldwide due in part to the distinct transformation of androgen-dependent prostate cancer (ADPC) into treatment-refractory castration-resistant prostate cancer (CRPC). Consequently, there is an urgent need to explore novel molecular mechanisms underlying treatment resistance in ADPC. Although numerous studies have alluded to the role of miR-200a in several cancers, the biological significance of miR-200a in prostate cancer remains unknown. After performing microarray analysis and reanalysis of the publicly available Memorial Sloan Kettering Cancer Center dataset, miR-200a expression was found higher in ADPC tissues and its expression was positively associated with survival of CRPC patients. In vitro studies showed that miR-200a overexpression in CRPC cells markedly suppressed cellular proliferation and facilitated apoptosis. In vivo studies indicated that overexpression of miR-200a inhibited growth and metastasis of prostate cancer. The luciferase reporter assay demonstrated that BRD4 is a direct target gene of miR-200a and it could reverse miR-200a-mediated biological effects in prostate cancer cells. Most importantly, our findings indicated that miR-200a suppresses the progression of CRPC by inhibiting the activation of BRD4-mediated AR signaling. This finding provides the foundation for the development of more personalized therapeutic approaches for CRPC patients.

BACKGROUND:Long non-coding RNA H19 polymorphisms were reported to be related to cancer susceptibility. However, the results from individual studies have been controversial or inconsistent. To clarify the associations between H19 single nucleotide polymorphisms (rs2107425, rs217727, rs2735971, rs2839698, rs3024270, and rs3741219) and the cancer susceptibility more accurately. METHODS:Relevant publications were searched from PubMed and EMBASE up to May 31, 2019, for studies in English only. The reference lists of the retrieved studies were also investigated. Pooled odds ratio (OR) with 95% confidence interval (CI) was calculated to find out the relationship between the H19 polymorphisms and cancer susceptibility. All of the data were analyzed using Stata 12.0. RESULTS:The results showed that rs2107425 polymorphisms was associated with an increasing cancer susceptibility in Asian (T vs C: OR 1.13, 95% CI 1.01-1.28; TT + CT vs CC: OR 1.21, 95% CI 1.03-1.44; CT vs CC: OR 1.21, 95% CI 1.01-1.44) and decreasing risk in Caucasian (T vs C: OR 0.90, 95% CI 0.84-0.97; TT + CT vs CC: OR 0.84, 95% CI 0.75-0.94; CT vs CC: OR 0.82, 95% CI 0.72-0.94). And rs217727 polymorphism was associated with an increasing cancer susceptibility in the Asian (A vs G: OR 1.09, 95% CI 1.02-1.17; AA + GA vs GG: OR 1.12, 95% CI 1.01-1.21; AA vs GG: OR 1.18, 95% CI 1.02-1.36). Additionally, rs2839698 polymorphism was associated with an increasing risk overall (A vs G: OR 1.18, 95% CI 1.06-1.31), in breast cancer (A vs G: OR 1.67, 95% CI 1.14-2.45; AA + AG vs GG: OR 1.98, 95% CI 1.20-3.25; AG vs GG: OR 1.89, 95% CI 1.16-3.07), in Asian (A vs G: OR 1.09, 95% CI 1.03-1.14; AA + AG vs GG: OR 1.11, 95% CI 1.04-1.21; AA vs AG + GG: OR 1.12, 95% CI 1.01-1.25; AA vs GG: OR 1.15, 95% CI 1.01-1.49; AG vs GG: OR 1.09, 95% CI 1.02-1.17), and in Caucasian (AA vs AG + GG: OR 1.81, 95% CI 1.25-2.61). CONCLUSION:H19 rs2107425, rs217727 and rs2839698 were associated with an increasing cancer susceptibility in Asian. Rs2107425 was associated with a decreasing risk and rs2839698 was associated with an increasing risk in Caucasian. No significant association was found in H19 rs2735971, rs3024270 and rs3741219 polymorphisms and cancer susceptibility.

Prostate cancer (PCa) is a frequently diagnosed malignant solid tumor in men. The etiology of PCa has been attributed to both environmental and genetic factors. In recent years, many studies have reported that miRNA gene single-nucleotide polymorphisms (SNPs) influence the susceptibility to several diseases such as cancer. To date, the mechanisms of PCa have remained unknown. The main aim of this study was to evaluate the association between PCa susceptibility and miRNA gene SNPs. A total of 156 PCa cases and 188 control subjects were included in this case-control study. The data were collected from hospitalized cases. We collected the demographic characteristic information, which included age, body mass index, tobacco smoking, alcohol consumption, and family history of cancer. Polymorphisms were analyzed by the ligase detection reaction. Unconditional logistic and stratified analyses were used to analyze the association between these SNPs and PCa susceptibility and to calculate the adjusted odds ratios (ORs) and the 95% confidence intervals (CIs). Cox regression model and the log-rank test were used to test the association between genetic variants and the overall survival. We found that miR-23a gene polymorphism rs3745453 carrying CC homozygotes had a 4.16-fold increased risk (95% CI =1.30-13.25) than those carrying the TT/CT genotypes (P= .02), and the C allele displayed a higher prevalence of PCa than the T allele (OR = 1.68, 95% CI =1.16-2.45, P= .01). Moreover, miR-23a showed that the homozygous carriers of the C-variant significantly increased the risk of survival rate as compared to the carriers of the TT/CT genotype (OR =9.67, 95% CI=2.83-33.09, P= .001). The rs3745453 polymorphism was potentially associated with PCa in the Chinese Han population and had an interactive relationship with the environmental factors.

BACKGROUND:Prostate cancer (PCa) is one of the most common malignant diseases among male patients. Although androgen deprivation therapy remains the main treatment for PCa, most patients would inevitably progress to castration-resistant PCa, which is the main cause of cancer-related deaths. Thus, novel antitumor agents are urgently needed. Recent studies demonstrated that aloperine (ALO) as a natural alkaloid showed antitumor effects in other cancer types. However, the biological function and underlying mechanisms of ALO in PCa have not been investigated.METHODS:PCa cell lines including LNCaP, PC3 and DU145 were cultured and treated with ALO. Cell Counting Kit-8 assay, colony formation assay, apoptosis assay and cell cycle assay were conducted to assess the biological role of ALO. In addition, a PCa subcutaneous xenograft mouse model was established to evaluate the role of ALO in terms of proliferation and apoptosis in vivo. We further measured the protein expression levels of p-Akt/Akt, p-ERK/ERK, c-Myc, cleaved caspase 3, p21, p53, Bcl-2 and Bax using the Western blot 48 h after ALO treatment of PCa cells.RESULTS:ALO effectively inhibited the cell viability of PCa by inducing cell cycle arrest via the activation of the p53/p21 pathway and triggering apoptosis in vitro and in vivo. ALO also inhibited phosphorylation of Akt and ERK protein kinases and activated cleaved caspase 3 while exerting antiproliferation function through inducing apoptosis and cell cycle arrest in PCa cells.CONCLUSION:Based on our findings, we conclude that ALO could suppress the tumor growth and promote cell apoptosis and cell cycle arrest in PCa cells, which indicated that ALO could act as a novel therapeutic agent in treatment of human PCa.

OBJECTIVE:To establish enzalutamide-resistant human prostate cancer cell lines and screen out the lncRNA and mRNA expression profiles associated with enzalutamide resistance.METHODS:Human prostate cancer cell lines LNCAP and C4-2B were cultured with 10 μmol/L enzalutamide for 6 months in vitro for the establishment of enzalutamide-resistant subclones LNCAP-ENZA and C4-2B-ENZA. The IC50 value and enzalutamide resistance index of each cell line were examined by MTT assay, the expressions of enzalutamide-related genes FL-AR, AR-V7 and HnRNPA1 were determined by Western blot, and the lncRNA and mRNA differential expressions of C4-2B and C4-2B-ENZA were detected by high-throughout lncRNA microarray.RESULTS:Compared with LNCAP and C4-2B, the IC50 values of enzalutamide-resistant subclones LNCAP-ENZA (60.83 μmol/L) and C4-2B-ENZA (88.32 μmol/L) were increased significantly (P < 0.05) and the enzalutamide-resistance indexes of the LNCAP-ENZA and C4-2B-ENZA cells were 4.94 and 4.67, respectively. The expressions of AR-V7 and HnRNPA1 were markedly up-regulated in the LNCAP-ENZA and C4-2B-ENZA cells as compared with those in the LNCAP and C4-2B cells, but that of FL-AR showed no significant change. A total of 1 440 lncRNAs and 1 236 mRNAs were identified as differentially expressed in the C4-2B-ENZA cells.CONCLUSIONS:Enzalutamide -resistant human prostate cancer cell subclones LNCAP-ENZA and C4-2B-ENZA were successfully established and enzalutamide resistance-associated lncRNA and mRNA were identified, which may provide some molecular evidence for the management of enzalutamide-resistant human prostate cancer.