Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(...Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(LncRNAs)participate in physiological and pathological processes through various epigenetic mechanisms.In this study,we found that a novel LncRNA named NKILA combined with multiple mucins and GALNTs potentially by several bioinformatics methods,and we used quantitative real-time PCR(RT-qPCR)to detect the expressions of NKILA,MUC5AC,MUC5B,and GALNT2 mRNA in 50 cases of asthma samples and 19 cases of normal samples,whose results showed that the expression of NKILA was significantly decreased in asthmatic samples,negatively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B,while GALNT2 was significantly increased in asthmatic tissues,and positively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B.In vitro,we used transient transfection technology to overexpress or interfere with NKILA and GALNT2 and then detected the expressions of MUC5AC and MUC5B via RT-qPCR and Western blot,which demonstrated GALNT2 can promote the expressions of MUC5AC and MUC5B protein,while NKILA could inhibit this effect.Furthermore,co-immunoprecipitation results showed that GALNT2 could bind to MUC5AC and MUC5B protein.RNA immunoprecipitation and RNA pull-down experiments showed that NKILA could bind to GALNT2.These evidences suggested that there are correlations among the expression of NKILA,GALNT2,MUC5AC,and MUC5B proteins in asthmatic patients.Mechanically,we concluded that NKILA can suppress the O-linked glycosylation of MUC5AC and MUC5B proteins by binding to GALNT2 and inhibit the expression of MUC5AC and MUC5B proteins.Our researches provided a potential therapeutic target for AHR.展开更多
<strong>Background:</strong> In the present study, autophagy-related long non-coding RNAs (lncRNAs) in lung adenocarcinoma (LUAD) were screened for diagnosis and prognosis, and the molecular mechanisms of ...<strong>Background:</strong> In the present study, autophagy-related long non-coding RNAs (lncRNAs) in lung adenocarcinoma (LUAD) were screened for diagnosis and prognosis, and the molecular mechanisms of LUAD at the genetic level were investigated. <strong>Methods:</strong> From The Cancer Genome Atlas (TCGA) database, 497 gene expression data and 436 clinical data of LUAD cases were collected for analysis. In addition, 232 autophagy-related genes (ARGs) were extracted from the Human Autophagy Database (HADb). Spearman rank correlation test and the Akaike information criterion (AIC) were performed to screen the data. After filtering, a survival model including three autophagy-related lncRNAs was generated. Based on the following formula: risk score = ΣCoef gene i×Gene i expression, the risk score of all LUAD patients could be calculated. LUAD patients were divided into two groups based on risk score for survival curve using Kaplan-Meier survival analysis. Both univariate and multivariate survival analyses were used to determine whether the three lncRNAs were independent prognostic factors using the survival package in R. Furthermore, the receiver operating characteristic (ROC) curves of clinical data were created to assess the stability of the survival model. Finally, the Gene Set Enrichment Analysis (GSEA) was used for analysis of related pathways. <strong>Results:</strong> A prognostic model consisting of three lncRNAs (AC011477.2, AC099850.3, and TRG-AS1) was generated for analysis. The 5-year survival rate in the high-risk group was 26.51% (95% CI: 0.1842 - 0.382), which was statistically lower than in the low-risk group 41.6% (95% CI: 0.307 - 0.563, P < 0.05). The area under the ROC curve (AUC) of risk score was 0.700, indicating a higher diagnostic accuracy of risk score. The results of GSEA showed enrichment in 36 pathways, including pyrimidine metabolism, pentose phosphate pathway, citric acid cycle, and cell cycle in the high-risk group, and FC-EPSILON-RI signal pathway, intestinal immune network produced by IgA, and ABC transporters in the low-risk group. <strong>Conclusion:</strong> The prognosis model composed of autophagy-related lncRNAs, AC011477.2, AC099850.3, and TRG-AS1, in LUAD can be used to predict the prognosis of LUAD patients and is expected to improve clinical treatment.展开更多
Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-me...Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-mesenchymal transition(EMT)exerts a vital role in tumor cell metastasis.However,it remains unclear whether long non-coding RNA(lncRNA)are implicated in EMT and influence ovarian cancer cell invasion and metastasis.This study was designed to investigate the impacts of lncRNA AC005224.4 on ovarian cancer.Methods:LncRNA AC005224.4,miR-140-3p,and snail family transcriptional repressor 2(SNAI2)expression levels in ovarian cancer and normal ovarian tissues were determined using real-time quantitative polymerase chain reaction(qRT-PCR).Cell Counting Kit-8(CCK-8)and Transwell(migration and invasion)assays were conducted to measure SKOV3 and CAOV-3 cell proliferation and metastasis.E-cadherin,N-cadherin,Snail,and Vimentin contents were detected using Western blot.Nude mouse xenograft assay was utilized to validate AC005224.4 effects in vivo.Dual-luciferase reporter gene assay confirmed the targeted relationship between miR-140-3p and AC005224.4 or SNAI2.Results:AC005224.4 and SNAI2 upregulation and miR-140-3p downregulation were observed in ovarian cancer tissues and cells.Silencing of AC005224.4 observably moderated SKOV3 and CAOV-3 cell proliferation,migration,invasion,and EMT process in vitro and impaired the tumorigenesis in vivo.miR-140-3p was a target of AC005224.4 and its reduced expression level was mediated by AC005224.4.miR-140-3p mimics decreased the proliferation,migration,and invasion of ovarian cancer cells.SNAI2 was identified as a novel target of miR-140-3p and its expression level was promoted by either AC005224.4 overexpression or miR-140-3p knockdown.Overexpression of SNAI2 also facilitated ovarian cancer cell viability and metastasis.Conclusion:AC005224.4 was confirmed as an oncogene via sponging miR-140-3p and promoted SNAI2 expression,contributing to better understanding of ovarian cancer pathogenesis and shedding light on exploiting the novel lncRNA-directed therapy against ovarian cancer.展开更多
文摘Glycosylation of mucins mediated by N-acetylgalactosaminyltransferases(GALNTs)is closely related to respiratory diseases such as asthma and chronic obstructive pulmonary disease(COPD).In addition,long non-coding RNAs(LncRNAs)participate in physiological and pathological processes through various epigenetic mechanisms.In this study,we found that a novel LncRNA named NKILA combined with multiple mucins and GALNTs potentially by several bioinformatics methods,and we used quantitative real-time PCR(RT-qPCR)to detect the expressions of NKILA,MUC5AC,MUC5B,and GALNT2 mRNA in 50 cases of asthma samples and 19 cases of normal samples,whose results showed that the expression of NKILA was significantly decreased in asthmatic samples,negatively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B,while GALNT2 was significantly increased in asthmatic tissues,and positively correlated with the severity of asthma and the expressions of MUC5AC and MUC5B.In vitro,we used transient transfection technology to overexpress or interfere with NKILA and GALNT2 and then detected the expressions of MUC5AC and MUC5B via RT-qPCR and Western blot,which demonstrated GALNT2 can promote the expressions of MUC5AC and MUC5B protein,while NKILA could inhibit this effect.Furthermore,co-immunoprecipitation results showed that GALNT2 could bind to MUC5AC and MUC5B protein.RNA immunoprecipitation and RNA pull-down experiments showed that NKILA could bind to GALNT2.These evidences suggested that there are correlations among the expression of NKILA,GALNT2,MUC5AC,and MUC5B proteins in asthmatic patients.Mechanically,we concluded that NKILA can suppress the O-linked glycosylation of MUC5AC and MUC5B proteins by binding to GALNT2 and inhibit the expression of MUC5AC and MUC5B proteins.Our researches provided a potential therapeutic target for AHR.
文摘<strong>Background:</strong> In the present study, autophagy-related long non-coding RNAs (lncRNAs) in lung adenocarcinoma (LUAD) were screened for diagnosis and prognosis, and the molecular mechanisms of LUAD at the genetic level were investigated. <strong>Methods:</strong> From The Cancer Genome Atlas (TCGA) database, 497 gene expression data and 436 clinical data of LUAD cases were collected for analysis. In addition, 232 autophagy-related genes (ARGs) were extracted from the Human Autophagy Database (HADb). Spearman rank correlation test and the Akaike information criterion (AIC) were performed to screen the data. After filtering, a survival model including three autophagy-related lncRNAs was generated. Based on the following formula: risk score = ΣCoef gene i×Gene i expression, the risk score of all LUAD patients could be calculated. LUAD patients were divided into two groups based on risk score for survival curve using Kaplan-Meier survival analysis. Both univariate and multivariate survival analyses were used to determine whether the three lncRNAs were independent prognostic factors using the survival package in R. Furthermore, the receiver operating characteristic (ROC) curves of clinical data were created to assess the stability of the survival model. Finally, the Gene Set Enrichment Analysis (GSEA) was used for analysis of related pathways. <strong>Results:</strong> A prognostic model consisting of three lncRNAs (AC011477.2, AC099850.3, and TRG-AS1) was generated for analysis. The 5-year survival rate in the high-risk group was 26.51% (95% CI: 0.1842 - 0.382), which was statistically lower than in the low-risk group 41.6% (95% CI: 0.307 - 0.563, P < 0.05). The area under the ROC curve (AUC) of risk score was 0.700, indicating a higher diagnostic accuracy of risk score. The results of GSEA showed enrichment in 36 pathways, including pyrimidine metabolism, pentose phosphate pathway, citric acid cycle, and cell cycle in the high-risk group, and FC-EPSILON-RI signal pathway, intestinal immune network produced by IgA, and ABC transporters in the low-risk group. <strong>Conclusion:</strong> The prognosis model composed of autophagy-related lncRNAs, AC011477.2, AC099850.3, and TRG-AS1, in LUAD can be used to predict the prognosis of LUAD patients and is expected to improve clinical treatment.
基金Xinjiang Uygur Autonomous Region Science and Technology Supporting Xinjiang Project(2017E0263)Tianjin Science and Technology Support Program Project(18YFZCSY00100)+3 种基金Program for New Century Excellent Talents in University in China(NCET-11-1066)Training Plan of leading subject talents in Tianjin colleges and universitiesthe National Natural Science Foundation of China(81972572)CAMS Innovation Fund for Medical Sciences(2016-I2M-1-001)
文摘Background:Ovarian cancer is one of the most widespread malignant diseases of the female reproductive system worldwide.The plurality of ovarian cancer is diagnosed with metastasis in the abdominal cavity.Epithelial-mesenchymal transition(EMT)exerts a vital role in tumor cell metastasis.However,it remains unclear whether long non-coding RNA(lncRNA)are implicated in EMT and influence ovarian cancer cell invasion and metastasis.This study was designed to investigate the impacts of lncRNA AC005224.4 on ovarian cancer.Methods:LncRNA AC005224.4,miR-140-3p,and snail family transcriptional repressor 2(SNAI2)expression levels in ovarian cancer and normal ovarian tissues were determined using real-time quantitative polymerase chain reaction(qRT-PCR).Cell Counting Kit-8(CCK-8)and Transwell(migration and invasion)assays were conducted to measure SKOV3 and CAOV-3 cell proliferation and metastasis.E-cadherin,N-cadherin,Snail,and Vimentin contents were detected using Western blot.Nude mouse xenograft assay was utilized to validate AC005224.4 effects in vivo.Dual-luciferase reporter gene assay confirmed the targeted relationship between miR-140-3p and AC005224.4 or SNAI2.Results:AC005224.4 and SNAI2 upregulation and miR-140-3p downregulation were observed in ovarian cancer tissues and cells.Silencing of AC005224.4 observably moderated SKOV3 and CAOV-3 cell proliferation,migration,invasion,and EMT process in vitro and impaired the tumorigenesis in vivo.miR-140-3p was a target of AC005224.4 and its reduced expression level was mediated by AC005224.4.miR-140-3p mimics decreased the proliferation,migration,and invasion of ovarian cancer cells.SNAI2 was identified as a novel target of miR-140-3p and its expression level was promoted by either AC005224.4 overexpression or miR-140-3p knockdown.Overexpression of SNAI2 also facilitated ovarian cancer cell viability and metastasis.Conclusion:AC005224.4 was confirmed as an oncogene via sponging miR-140-3p and promoted SNAI2 expression,contributing to better understanding of ovarian cancer pathogenesis and shedding light on exploiting the novel lncRNA-directed therapy against ovarian cancer.
