LncRNA RP3-326I13.1 Promoted Cisplatin Resistance of Lung Adenocarcinoma by Collaborating RNA Binding Protein HSP90B and Upregulating Downstream Molecule MMP13 

2020 ◽  
Author(s):  
Huixin Zhou ◽  
Shihao Xu ◽  
Wenjing Shi ◽  
Xiaolu Huang ◽  
Jie Chen ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Nude Mice ◽  
Rna Binding ◽  
Cisplatin Resistance ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
A549 Cells ◽  
Downstream Target

Abstract Background:We previously obtained a lncRNA RP3-326I13.1, which significantly upregulated by cisplatin resistance in lung adenocarcinoma (LAD), but the biological function and molecular mechanism is unclear. Methods:Expression levels of RP3-326I13.1 and HSP90B mRNA were estimated by qPCR from 57 pairs of LAD and NT samples without and with cisplatin. Knockdown and overexpression in A549/DDP and A549 cell lines by lentiviral- mediated techniques to observe changes in tumor behavior in A549/DDP and A549 cells, as well as tumorigenicity in experimental nude mice. The ranscriptome was sequenced to obtain downstream target molecules of RP3-326I13.1 and RNA-binding proteins were obtained using RNA pulldown. Results: QPCR showed that the expression level of RP3-326I13.1 and HSP90B mRNA in A549/DDP cells, LAD tissues and progressive LAD tissues (cisplatin treatment was not effective) were tangibly higher than that of A549 cells, adjacent tissues, and complete remission (P=0.0037, P=0.0181; P=0.0027, P=0.009 and P=0.002, P=0.007). RP3-326I13.1 markedly enhanced the proliferation, migrate, invasion, clonal proliferation ability of LAD cell lines and speed and weight of tumorigenicity in nude mice experiment while increased the proportion of G1 phase cells (P=0.019). RNA-pull down and mass spectrometry obtained RNA binding protein HSP90B and HSP90B clearly decreased proliferation, invasive ability while increased the apoptosis of LAD cell lines after knocked down. We found matrix metalloproteinase-13 (MMP-13) was RP3-326I13.1 downstream target gene. Conclusions: So, RP3-326I13.1 was a drug-resistant relative lncRNA promoted cisplatin resistance of lung adenocarcinoma by collaborating RNA binding protein HSP90B and upregulating downstream target molecule MMP13.

LncRNA KCNK15-AS1 inhibits pancreatic cancer progression by promoting the expression of its binding protein ACTR3B

2020 ◽  
Author(s):  
Pengfei Wu ◽  
Hao Yuan ◽  
Xiangya Ding ◽  
Qun Chen ◽  
Wanli Ge ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Pancreatic Cancer ◽  
Cell Growth ◽  
Cyclin D1 ◽  
Cell Lines ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Luciferase Reporter

Abstract Background LncRNAs are reported to play an essential role in multiple tumors, including pancreatic cancer. LncRNAs could impact tumor growth via RNA-binding proteins, working as a coactivators of transcription factors or impacting their gene expression via posttranscriptional regulation. Our study aimed to elucidate the function and mechanism of lncRNA KCNK15-AS1 and its binding protein ACTR3B in PC progression. Our previous data indicated that KCNK15-AS1 is downregulated in PC tissues and cell lines compared to normal controls. Methods In this study, we overexpressed KCNK15-AS1 and ACTR3B in both BxPC-3 and Mia-PaCa-2 cells to detect the cellular phenotype in vitro and in vivo. RNA pulldown assays, mass spectrometry assays and RNA-binding protein immunoprecipitation assays were used to verify KCNK15-AS1 RNA binding protein ACTR3B. Luciferase reporter assay and ubiquitination assay were proceeded to detect the mechanism KCNK15-AS1 upregulated ACTR3B expression. Results Our results showed that overexpression of KCNK15-AS1 significantly inhibited the proliferation, colony formation and migration of PC cells. ACTR3B was screened by RNA pulldown and mass spectrometry assays. RNA-binding protein immunoprecipitation assays confirmed that KCNK15-AS1 physically bound to ACTR3B. Furthermore, mechanistic analyses demonstrated that KCNK15-AS1 promoted ACTR3B expression by inhibiting ACTR3B ubiquitin-mediated degradation and enhancing its promoter activity. Additionally, ACTR3B presented low expression in PC tissues and cell lines, and PC cell growth was significantly repressed when ACTR3B was overexpressed. Moreover, knockdown of ACTR3B in KCNK15-AS1-overexpressing cells reversed the effects of KCNK15-AS1 on PC cell growth via the cyclin D1/CDK4 axis. Conclusion Briefly, our study indicated that the lncRNA KCNK15-AS1/ACTR3B/cyclin D1/CDK4 axis may inhibit PC progression, which provides a potential therapeutic target for PC.

scholarly journals Mechanistic study of lncRNA UCA1 promoting growth and cisplatin resistance in lung adenocarcinoma

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jiali Fu ◽  
Jingjing Pan ◽  
Xiang Yang ◽  
Yan Zhang ◽  
Fanggui Shao ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Rna Binding ◽  
Cisplatin Resistance ◽  
Excision Repair ◽  
Quantitative Polymerase Chain Reaction ◽  
A549 Cells ◽  
Expression Level ◽  
Nuclear Antigen ◽  
Mechanistic Study

Abstract Aim This study aimed to explore the mechanism of LncRNA urothelial carcinoma-associated 1 (UCA1) promoting cisplatin resistance in lung adenocarcinoma (LUAD). Method The UCA1 expression level in LUAD cell lines was detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). We overexpressed UCA1 in A549 cells and downregulated UCA1 in A549/DDP cells by the lentivirus‑mediated technique. Subsequently, in vitro, and in vivo functional experiments were performed to investigate the functional roles of UCA1 in the growth and metastasis of LUAD cell lines. Furthermore, RNA pulldown, mass spectrometry, and RNA immunoprecipitation technique were performed to analyze various downstream target factors regulated by UCA1. Results The results revealed a higher UCA1 expression level in A549/DDP cells and LUAD tissues than in A549 cells and adjacent cancer tissues. UCA1 expression was significantly associated with distant metastasis, clinical stage, and survival time of patients with LUAD. UCA1 overexpression significantly increased the proliferation, invasion, clone formation, and cisplatin resistance ability and enhanced the expression levels of proliferating cell nuclear antigen and excision repair cross-complementing gene 1 in A549 cells. However, these trends were mostly reversed after the knockdown of UCA1 in A549/DDP cells. Tumorigenic assays in nude mice showed that UCA1 knockdown significantly inhibited tumor growth and reduced cisplatin resistance. Enolase 1 was the RNA-binding protein (RBP) of UCA1. Conclusion Based on the results, we concluded that UCA1 promoted LUAD progression and cisplatin resistance and hence could be a potential diagnostic marker and therapeutic target in patients with LUAD.

scholarly journals LncRNA RP3-326I13.1 Promoted Cisplatin Resistance of Lung Adenocarcinoma by Collaborating RNA Binding Protein HSP90B and Upregulating Downstream Molecule MMP13

2021 ◽  
Keyword(s):  
Lung Adenocarcinoma ◽  
Full Text ◽  
Rna Binding ◽  
Cisplatin Resistance ◽  
Binding Protein ◽  
Rna Binding Protein

Abstract The full text of this preprint has been withdrawn by the authors due to author disagreement with the posting of the preprint. Therefore, the authors do not wish this work to be cited as a reference. Questions should be directed to the corresponding author.

scholarly journals Predicting the Clinical Outcome of Lung Adenocarcinoma Using a Novel Gene Pair Signature Related to RNA-Binding Protein

2020 ◽  
Vol 2020 ◽  
pp. 1-14
Author(s):  
Liangliang Meng ◽  
Xiaoxi He ◽  
Xiao Zhang ◽  
Xiaobo Zhang ◽  
Yingtian Wei ◽  
...  
Keyword(s):  
T Cells ◽  
Lung Adenocarcinoma ◽  
Gene Pair ◽  
Prognostic Model ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Risk Group ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
High Risk Group

Adenocarcinoma is the most common type of lung cancer, and patients have varying prognoses. RNA-binding proteins (RBP) are deemed to be closely associated with tumorigenesis and development, but the exact mechanism is currently unknown. This study was aimed at constructing a new robust prognostic model based on RNA-binding protein-related gene pair scores for better clinical guidance. The model for this study was constructed based on data of lung adenocarcinoma from The Cancer Genome Atlas (TCGA) database. Prognosis-related RBP gene pair models were created based on differentially expressed genes, and the accuracy of the models was verified in a different age, staging, and other subdatasets. A total of 379 RNA-binding protein-related genes were differentially expressed in tumor tissue. From these genes, we constructed a prognostic model consisting of 33 gene pairs, which were found to be significantly associated with survival in TCGA dataset ( P < 0.0001 , hazard ratio   HR = 4.380 (3.139 to 6.111)) and different subdatasets. As expected, the results were verified in the GEO validation cohort ( P = 7.8 × 10 − 3 , HR = 1.597 (1.095 to 2.325)). We found that the signature exhibited an independent prognostic factor in both the univariate and multivariate Cox regression analyses ( P < 0.001 ). CIBERSORT was applied to estimate the fractions of infiltrated immune cells in bulk tumor tissues. CD8 T cells, activated dendritic cells, regulatory T cells (Tregs), and activated CD4 memory T cells presented a significantly lower fraction in the high-risk group ( P < 0.01 ). Patients in the high-risk group had significantly higher tumor mutational burden (TMB) ( P = 4.953 e − 04 ) and lower levels of immune cells ( P = 3.473 e − 05 ) and stromal cells ( P = 0.005 ) in the tumor microenvironment than those in the low-risk group. Furthermore, the Protein-protein interaction (PPI) network and various enrichment analyses have genuinely uncovered the interrelationships and potential functions of the RBP genes within the model. The results of the present study validated the importance of RNA-binding proteins in tumorigenesis and progression and support the RBP gene-related signature as a promising marker for prognosis prediction in lung adenocarcinoma.

scholarly journals The Interplay of RNA Binding Proteins, Oxidative Stress and Mitochondrial Dysfunction in ALS

Antioxidants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 552
Author(s):  
Jasmine Harley ◽  
Benjamin E. Clarke ◽  
Rickie Patani
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Mitochondrial Dysfunction ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Therapeutic Strategies ◽  
Lateral Sclerosis

RNA binding proteins fulfil a wide number of roles in gene expression. Multiple mechanisms of RNA binding protein dysregulation have been implicated in the pathomechanisms of several neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Oxidative stress and mitochondrial dysfunction also play important roles in these diseases. In this review, we highlight the mechanistic interplay between RNA binding protein dysregulation, oxidative stress and mitochondrial dysfunction in ALS. We also discuss different potential therapeutic strategies targeting these pathways.

scholarly journals The RNA-binding protein ELAVL1/HuR is essential for mouse spermatogenesis, acting both at meiotic and postmeiotic stages

2011 ◽  
Vol 22 (16) ◽  
pp. 2875-2885 ◽  
Author(s):  
Mai Nguyen Chi ◽  
Jacques Auriol ◽  
Bernard Jégou ◽  
Dimitris L. Kontoyiannis ◽  
James M.A. Turner ◽  
...  
Keyword(s):  
Germ Cells ◽  
Posttranscriptional Regulation ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Female Sterility ◽  
Targeted Deletion

Posttranscriptional mechanisms are crucial to regulate spermatogenesis. Accurate protein synthesis during germ cell development relies on RNA binding proteins that control the storage, stability, and translation of mRNAs in a tightly and temporally regulated manner. Here, we focused on the RNA binding protein Embryonic Lethal Abnormal Vision (ELAV) L1/Human antigen R (HuR) known to be a key regulator of posttranscriptional regulation in somatic cells but the function of which during gametogenesis has never been investigated. In this study, we have used conditional loss- and gain-of-function approaches to address this issue in mice. We show that targeted deletion of HuR specifically in germ cells leads to male but not female sterility. Mutant males are azoospermic because of the extensive death of spermatocytes at meiotic divisions and failure of spermatid elongation. The latter defect is also observed upon HuR overexpression. To elucidate further the molecular mechanisms underlying spermatogenesis defects in HuR-deleted and -overexpressing testes, we undertook a target gene approach and discovered that heat shock protein (HSP)A2/HSP70-2, a crucial regulator of spermatogenesis, was down-regulated in both situations. HuR specifically binds hspa2 mRNA and controls its expression at the translational level in germ cells. Our study provides the first genetic evidence of HuR involvement during spermatogenesis and reveals Hspa2 as a target for HuR.

scholarly journals Development and validation of a prediction model for lung adenocarcinoma based on RNA-binding protein

2021 ◽  
Vol 9 (6) ◽  
pp. 474-474
Author(s):  
Longjun Yang ◽  
◽  
Rusi Zhang ◽  
Guangran Guo ◽  
Gongming Wang ◽  
...  
Keyword(s):  
Prediction Model ◽  
Lung Adenocarcinoma ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Development And Validation

scholarly journals Investigating the Roles of RNA Binding Protein Combinations in Neuronal Function and Organismal Behavior

2019 ◽  
Vol 4 (Spring 2019) ◽  
Author(s):  
Alexa Vandenburg
Keyword(s):  
Binding Sites ◽  
Neuronal Development ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Wild Type ◽  
C Elegans ◽  
Neuronal Gene ◽  
Touch Response

The Norris lab recently identified two RNA binding proteins required for proper neuron-specific splicing. The lab conducted touch- response behavioral assays to assess the function of these proteins in touch-sensing neurons. After isolating C. elegans worms with specific phenotypes, the lab used automated computer tracking and video analysis to record the worms’ behavior. The behavior of mutant worms differed from that of wild-type worms. The Norris lab also discovered two possible RNA binding protein sites in SAD-1, a neuronal gene implicated in the neuronal development of C. elegans1. These two binding sites may control the splicing of SAD-1. The lab transferred mutated DNA into the genome of wild-type worms by injecting a mutated plasmid. The newly transformed worms fluoresced green, indicating that the two binding sites control SAD-1 splicing.

PUB1 is a major nuclear and cytoplasmic polyadenylated RNA-binding protein in Saccharomyces cerevisiae

1993 ◽  
Vol 13 (10) ◽  
pp. 6102-6113
Author(s):  
J T Anderson ◽  
M R Paddy ◽  
M S Swanson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Uv Light ◽  
Binding Protein ◽  
Consensus Sequence ◽  
Rna Binding Protein ◽  
Polyadenylated Rna ◽  
Polyadenylated Rnas

Proteins that directly associate with nuclear polyadenylated RNAs, or heterogeneous nuclear RNA-binding proteins (hnRNPs), and those that associate with cytoplasmic mRNAs, or mRNA-binding proteins (mRNPs), play important roles in regulating gene expression at the posttranscriptional level. Previous work with a variety of eukaryotic cells has demonstrated that hnRNPs are localized predominantly within the nucleus whereas mRNPs are cytoplasmic. While studying proteins associated with polyadenylated RNAs in Saccharomyces cerevisiae, we discovered an abundant polyuridylate-binding protein, PUB1, which appears to be both an hnRNP and an mRNP. PUB1 and PAB1, the polyadenylate tail-binding protein, are the two major proteins cross-linked by UV light to polyadenylated RNAs in vivo. The deduced primary structure of PUB1 indicates that it is a member of the ribonucleoprotein consensus sequence family of RNA-binding proteins and is structurally related to the human hnRNP M proteins. Even though the PUB1 protein is a major cellular polyadenylated RNA-binding protein, it is nonessential for cell growth. Indirect cellular immunofluorescence combined with digital image processing allowed a detailed comparison of the intracellular distributions of PUB1 and PAB1. While PAB1 is predominantly, and relatively uniformly, distributed within the cytoplasm, PUB1 is localized in a nonuniform pattern throughout both the nucleus and the cytoplasm. The cytoplasmic distribution of PUB1 is considerably more discontinuous than that of PAB1. Furthermore, sucrose gradient sedimentation analysis demonstrates that PAB1 cofractionates with polyribosomes whereas PUB1 does not. These results suggest that PUB1 is both an hnRNP and an mRNP and that it may be stably bound to a translationally inactive subpopulation of mRNAs within the cytoplasm.

scholarly journals A potential role for a novel ZC3H5 complex in regulating mRNA translation in Trypanosoma brucei

2020 ◽  
Vol 295 (42) ◽  
pp. 14291-14304
Author(s):  
Kathrin Bajak ◽  
Kevin Leiss ◽  
Christine Clayton ◽  
Esteban Erben
Keyword(s):  
Gene Expression ◽  
Trypanosoma Brucei ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Binding Protein ◽  
Affinity Purification ◽  
Critical Role ◽  
Rna Binding Protein ◽  
Mrna Translation

In Trypanosoma brucei and related kinetoplastids, gene expression regulation occurs mostly posttranscriptionally. Consequently, RNA-binding proteins play a critical role in the regulation of mRNA and protein abundance. Yet, the roles of many RNA-binding proteins are not understood. Our previous research identified the RNA-binding protein ZC3H5 as possibly involved in gene repression, but its role in controlling gene expression was unknown. We here show that ZC3H5 is an essential cytoplasmic RNA-binding protein. RNAi targeting ZC3H5 causes accumulation of precytokinetic cells followed by rapid cell death. Affinity purification and pairwise yeast two-hybrid analysis suggest that ZC3H5 forms a complex with three other proteins, encoded by genes Tb927.11.4900, Tb927.8.1500, and Tb927.7.3040. RNA immunoprecipitation revealed that ZC3H5 is preferentially associated with poorly translated, low-stability mRNAs, the 5′-untranslated regions and coding regions of which are enriched in the motif (U/A)UAG(U/A). As previously found in high-throughput analyses, artificial tethering of ZC3H5 to a reporter mRNA or other complex components repressed reporter expression. However, depletion of ZC3H5 in vivo caused only very minor decreases in a few targets, marked increases in the abundances of very stable mRNAs, an increase in monosomes at the expense of large polysomes, and appearance of “halfmer” disomes containing two 80S subunits and one 40S subunit. We speculate that the ZC3H5 complex might be implicated in quality control during the translation of suboptimal open reading frames.

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