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 Mechanistic study of LncRNA UCA1 promoting growth and cisplatin resistance in lung adenocarcinoma

2021 ◽  
Author(s):  
Jiali Fu ◽  
Jingjing Pan ◽  
Xiang Yang ◽  
Yan Zhang ◽  
Fanggui Shao ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Nude Mice ◽  
A549 Cell ◽  
Rna Binding ◽  
Cisplatin Resistance ◽  
Quantitative Polymerase Chain Reaction ◽  
Tumor Formation ◽  
The Cancer Genome Atlas ◽  
Expression Level ◽  
Mechanistic Study

Abstract Background: The main objective of the current research was to explore the mechanism of LncRNA UCA1 promoting cisplatin resistance in lung adenocarcinoma (LUAD).Method: The UCA1 expression level of LUAD cell lines was herein detected by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). We overexpressed UCA1 in A549 cell and downregulated UCA1 in A549/DDP cell by lentivirus‑mediated technique. We analyzed their biological differences by cell function experiments, RNA pulldown, protein mass spectrometry (MS), and RNA immunoprecipitation technique (RIP). Tumor formation in nude mice was used to investigate the effect of UCA1 on the proliferation and cisplatin sensitivity of A549/DDP in vivo.Result: The results revealed that a higher expression level of UCA1 was expressed in the A549/DDP cell and LUAD tissues than that in A549 cell and adjacent cancer tissues. UCA1 was significantly associated with M stage and clinical stage of LUAD patients from The Cancer Genome Atlas (TCGA) database. Patients with high UCA1 expression had a shorter survival time. After UCA1 overexpressed, the cells capability of proliferation, migration, invasion, clone formation, cisplatin resistance, and the expression level of proteins related to proliferation and drug resistance PCNA, ERCC1 were enhanced, while these trends were mostly reversed in the cells knockdown with UCA1 expression. Tumorigenic assays in nude mice showed that knockdown of UCA1 significantly inhibited tumor growth and reduced cisplatin resistance. It confirmed Enolase 1 (ENO1) was one of RNA binding protein of UCA1.Conclusion: Based on these results, we concluded that UCA1 promoted LUAD progression and cisplatin resistance by binding ENO1 and UCA1 could be a potential diagnostic marker and therapeutic target of LUAD patients.

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.

scholarly journals Akt kinase LANCL2 functions as a key driver in EGFR-mutant lung adenocarcinoma tumorigenesis

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Yuqing Lou ◽  
Jianlin Xu ◽  
Yanwei Zhang ◽  
Wei Zhang ◽  
Xueyan Zhang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Cell Line ◽  
Quantitative Polymerase Chain Reaction ◽  
Mutant Cell ◽  
A549 Cells ◽  
The Cancer Genome Atlas ◽  
Akt Kinase ◽  
Egfr Expression

AbstractEpidermal growth factor receptor (EGFR) is a key oncogene in lung adenocarcinoma (LUAD). Resistance to EGFR tyrosine kinase inhibitors is a major obstacle for EGFR-mutant LUAD patients. Our gene chip array, quantitative polymerase chain reaction validation, and shRNA-based high-content screening identified the Akt kinase lanthionine synthetase C-like protein 2 (LANCL2) as a pro-proliferative gene in the EGFR-mutant LUAD cell line PC9. Therefore, we investigated whether LANCL2 plays a role in promoting cell proliferation and drug resistance in EGFR-mutant LUAD. In silico clinical correlation analysis using the Cancer Genome Atlas Lung Adenocarcinoma dataset revealed a positive correlation between LANCL2 and EGFR expression and an inverse relationship between LANCL2 gain-of-function and survival in LUAD patients. The EGFR-mutant LUAD cell lines PC9 and HCC827 displayed higher LANCL2 expression than the non-EGFR-mutant cell line A549. In addition, LANCL2 was downregulated following gefitinib+pemetrexed combination therapy in PC9 cells. LANCL2 knockdown reduced proliferation and enhanced apoptosis in PC9, HCC827, and A549 cells in vitro and suppressed murine PC9 xenograft tumor growth in vivo. Notably, LANCL2 overexpression rescued these effects and promoted gefitinib + pemetrexed resistance in PC9 and HCC827 cells. Pathway analysis and co-immunoprecipitation followed by mass spectrometry of differentially-expressed genes in LANCL2 knockdown cells revealed enrichment of several cancer signaling pathways. In addition, Filamin A and glutathione S-transferase Mu 3 were identified as two novel protein interactors of LANCL2. In conclusion, LANCL2 promotes tumorigenic proliferation, suppresses apoptosis, and promotes gefitinib+pemetrexed resistance in EGFR-mutant LUAD cells. Based on the positive association between LANCL2, EGFR, and downstream Akt signaling, LANCL2 may be a promising new therapeutic target for EGFR-mutant LUAD.

scholarly journals TEK Suppresses Lung Adenocarcinoma Cell Phenotypes by Interacting with miR-19a-3p

2020 ◽  
Author(s):  
Tao Peng ◽  
Fan Yang ◽  
Zhanwen Sun ◽  
Jie Yan
Keyword(s):  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
A549 Cells ◽  
Significant Inhibition ◽  
Adhesion Assay ◽  
Novel Therapy ◽  
Adenocarcinoma Cell ◽  
Protein Levels ◽  
Apoptosis Assay ◽  
Cell Phenotypes

Abstract Background: We identified TEK as a key gene that that participates in lung adenocarcinoma cell migration and adhesion, and miR-19a-3p a potential upstream regulator of TEK. Both TEK and miR-19a-3p have been reported during lung cancer development. However, how TEK/miR-19a-3p interactome regulates lung adenocarcinoma remains unraveled. We herein aim to report a novel TEK/miR-19a-3p interactome in lung adenocarcinoma.Methods: The mRNA and protein expression of TEK in tissues and cell lines were determined using qPCR and Western blot, respectively. CCK-8 assay, EDU assay, flow-cytometry cell apoptosis assay, scratch assay, and cell-to-extracellular matrix adhesion assay were performed to detect the proliferation, apoptosis, migration, and adhesion of A549 and H1975 cell lines. Results：Both mRNA and protein levels of TEK were down-regulated in tumor tissues and cell lines. Compared with the control, the transfection of TEK overexpression plasmids into H1975 and A549 cells led to the significant inhibition of cancerous phenotypes. On the other hand, miR-19a-3p promoted lung adenocarcinoma cancerous cell phenotypes by downregulating TEK.Conclusions: TEK can be a potential LUAD tumor suppressor by interacting with miR-19a-3p. This novel interactome can be used as a novel therapy target for LUAD.

scholarly journals Comprehensive analysis of circular RNA expression profiles in cisplatin-resistant non-small cell lung cancer cell lines

2020 ◽  
Vol 52 (9) ◽  
pp. 944-953
Author(s):  
Lin Song ◽  
Zhilei Cui ◽  
Xuejun Guo
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Cell Lines ◽  
Cell Lung Cancer ◽  
Kinase Activity ◽  
Cisplatin Resistance ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Small Cell ◽  
Small Cell Lung

Abstract Platinum-based drugs such as cisplatin are widely used in combination chemotherapy for non-small cell lung cancer (NSCLC) owing to their high clinical response rate; however, acquired resistance to cisplatin is eventually inevitable. Circular RNAs (circRNAs) are involved in the development of diverse types of cancers, but their connection to cisplatin-resistance in NSCLC has not been studied. In the present study, two cisplatin-resistant NSCLC cell lines (A549/DDP and PC9/DDP) were established by gradually increasing concentrations of cisplatin in the media. The resulting cell lines possessed high resistance to cisplatin and strong proliferation, migration, and colony formation abilities compared to the parental cells. Microarray analysis identified 19,161 circRNAs that were dysregulated in cisplatin-resistant cell lines (fold change abs>2), including 11,915 up-regulated and 7246 down-regulated circRNAs. The expression of the top five up-regulated and down-regulated circRNAs was validated using real-time quantitative polymerase chain reaction. A circRNA–micro RNA (miRNA) network of the top 20 dysregulated circRNAs and their predicted miRNAs was constructed using Cytoscape. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that the host genes of the identified circRNAs were involved in the regulation of MAP kinase kinase kinase kinase activity, 6-phosphofructo-2-kinase activity, focal adhesion, ErbB signaling, and ECM-receptor interactions, which may contribute to cisplatin resistance in NSCLC. In summary, this is the first report on circRNA profiling in cisplatin-resistant NSCLC cells and it provides new potential targets for the reversal of cisplatin resistance in NSCLC.

Integration of mRNA and microRNA profiles as prognostic and predictive markers in lung adenocarcinoma

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 7522-7522
Author(s):  
J. H. Strickler ◽  
W. Mostertz ◽  
W. Kim ◽  
K. Walters ◽  
M. Stevenson ◽  
...  
Keyword(s):  
High Risk ◽  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Cisplatin Resistance ◽  
Early Stage ◽  
Gene Signature ◽  
Discovery Cohort ◽  
Risk Of Recurrence ◽  
Increased Risk

7522 Background: Lung adenocarcinoma (ADC) is a distinct biologic entity with unique gene amplifications (Weir B, Nature 2008). Yet, comprehensive transcriptomic analysis, including microRNAs, specific to lung ADC are lacking. Methods: Using mRNA expression data from a discovery cohort of 154 patients with histologically proven early stage (I and II) lung ADC, signatures of oncogenic pathway and tumor microenvironment status were applied and further organized by hierarchical clustering to develop a metagene model. Further, using in vitro assays in a large cohort of lung ADC cell lines (n = 42) with corresponding mRNA and microRNA data, novel microRNAs associated with a poor prognosis and their relationship to cisplatin resistance was elucidated. Results: In the discovery cohort of 154 patients with early stage disease, activation of oncogenic pathways associated with wound healing (angiogenesis), chromosomal instability, and STAT signaling were associated with an increased risk of recurrence (p<0.001). Utilizing the extremes of survival to identify cohorts of patients as high and low risk phenotypes, using bayesian regression, a 100 gene signature (‘metagene') that captured the diversity of signaling pathways unique to patients at increased risk of recurrence was identified and validated in an independent cohort (n = 364) of lung ADC samples with 78.3% accuracy. Kaplan Meier survival analysis and multivariate analysis further confirmed the independent prognostic value of the 100 gene signature (p= 0.007). Using in vitro cell proliferation assays, predicted high risk lung ADC cell lines were identified as being more resistant to cisplatin therapy than those predicted to be low risk (p=0.001). In a novel manner, we also identified several microRNAs (miR-215, miR-98, miR- 643, let-7b, miR-665, miR-629) associated with a high risk of recurrence and more importantly cisplatin resistance. Conclusions: mRNA and microRNA profiles reflect unique aspects of individual tumors and may characterize histology-specific tumor heterogeneity in lung ADC, providing an opportunity to better characterize the oncogenic process and refine therapeutic options. No significant financial relationships to disclose.

scholarly journals Autorepression of Epstein-Barr Virus Nuclear Antigen 1 Expression by Inhibition of Pre-mRNA Processing

2007 ◽  
Vol 82 (4) ◽  
pp. 1679-1687 ◽  
Author(s):  
Mikio Yoshioka ◽  
Michelle M. Crum ◽  
Jeffery T. Sample
Keyword(s):  
Cell Lines ◽  
Epstein Barr Virus ◽  
Rna Binding ◽  
Cytotoxic T Cells ◽  
Nuclear Antigen ◽  
Genome Maintenance ◽  
Barr Virus ◽  
Latently Infected ◽  
Infected Cells ◽  
Epstein Barr

ABSTRACT Epstein-Barr virus (EBV) latent infection, and its associated oncogenic potential, is dependent on genome maintenance functions of EBV nuclear antigen 1 (EBNA-1), one of six EBNAs expressed from a common promoter (Wp and then Cp) upon infection of naive B cells. Subsequent host-mediated silencing, however, necessitates the expression of EBNA-1 from the EBNA-1-specific promoter Qp to ensure against genome loss during cell division, including EBV-associated malignancy. Here we addressed the mechanism by which EBNA-1 represses Qp through binding downstream of the transcription start site and the role of this autoregulatory function in EBV latency. Our results revealed that EBNA-1 does not inhibit transcription from Qp, as previously predicted, but acts post- or cotranscriptionally to block the processing of primary transcripts. This does not, however, require the RGG motifs responsible for strong but nonspecific RNA binding by EBNA-1. Within isogenic B-cell lines using either Cp/Wp or Qp, EBNA-1 occupancy of Qp is equivalent, suggesting that autoregulation occurs, albeit to different degrees, during full and restricted EBV latency programs. Finally, in cell lines using Cp or Wp for EBNA expression, unprocessed transcripts from Qp are detectable in the absence of corresponding mRNAs, providing further evidence that this novel mechanism of EBNA-1 action functions during latency. This posttranscriptional mechanism of regulation would provide an efficient means to monitor and regulate EBNA-1 expression from Qp, ensuring levels adequate for genome maintenance but, perhaps more importantly, below an immunogenic threshold above which latently infected cells may be at risk for elimination by EBNA-1-specific cytotoxic T cells.

scholarly journals 1-Hydroxy-8-methoxy-anthraquinon reverses cisplatin resistance by inhibiting 6PGD in cancer cells

2019 ◽  
Vol 14 (1) ◽  
pp. 454-461 ◽  
Author(s):  
Huamin Zhang ◽  
Haowei Zhang ◽  
Sihui Wang ◽  
Zhihai Ni ◽  
Tiejun Wang
Keyword(s):  
Lung Cancer ◽  
Cell Lines ◽  
Small Molecule ◽  
Cisplatin Resistance ◽  
Small Molecule Inhibitor ◽  
Resistant Cell ◽  
Expression Level ◽  
Sensitive Cell ◽  
Molecule Inhibitor ◽  
Resistant Cells

AbstractTargeting 6-phosphogluconate dehydrogenase (6PGD) can inhibit cancer cell proliferation and tumor growth. However, the relationship between 6PGD and cisplatin resistance still needs further study. Cisplatin-sensitive and cisplatin-resistant ovarian cancer OV2008 and C13* lines and lung cancer A549 and A549DDP lines were treated with different concentrations of cisplatin and cell viability was evaluated. We also compared the growth rates and the cell cycle distributions between cisplatin-sensitive and cisplatin-resistant cells. The expression level of 6PGD was detected by immunoblotting. The Chou-Talalay method was used to evaluate the effect of a combination treatment using cisplatin and the small molecule inhibitor 1-Hydroxy-8-methoxy-anthraquinon (S3) that targets 6PGD. The cisplatin-resistant ovarian and lung cancer cell lines grew faster than the cisplatin- sensitive cell lines, with more cells in S and G2 phases in cisplatin-resistant cell lines. The expression level of 6PGD in cisplatin-resistant cell lines was significantly increased compared with cisplatin-sensitive cell lines. Furthermore, treatment of cells with the S3 small molecule inhibitor of 6PGD together with cisplatin could overcome cisplatin resistance. The expression level of 6PGD in cisplatin-resistant cells lines was significantly upregulated, and the resistance to cisplatin of drug-resistant cells lines could be overcome when treated with the small molecule inhibitor S3 that specifically targets 6PGD.

scholarly journals m6A RNA Methylation Regulators Act as Potential Prognostic Biomarkers in Lung Adenocarcinoma

2021 ◽  
Vol 12 ◽  
Author(s):  
Hongbo Wang ◽  
Xiangxuan Zhao ◽  
Zaiming Lu
Keyword(s):  
Lung Adenocarcinoma ◽  
Rna Binding ◽  
Excision Repair ◽  
Rna Degradation ◽  
High Risk Group ◽  
Migration And Invasion ◽  
Binding Motif ◽  
Regulatory Factors ◽  
Heterogeneous Nuclear Ribonucleoprotein ◽  
Nuclear Ribonucleoprotein

N6-methyladenosine [m(6)A/m6A] methylation is one of the most common RNA modifications in eukaryotic cell mRNA and plays an important regulatory role in mRNA metabolism, splicing, translocation, stability, and translation. Previous studies have demonstrated that the m6A modification is highly associated with tumor cell proliferation, migration, and invasion. In the present study, five m6A regulatory factors have been revealed, namely heterogeneous nuclear ribonucleoprotein A2/B1(HNRNPA2B1), heterogeneous nuclear ribonucleoprotein C (HNRNPC), Vir like m6A methyltransferase associated protein (KIAA1429/VIRMA), RNA binding motif protein 15 (RBM15) and methyltransferase like 3 (METTL3), which are closely related to the overall survival (OS) of patients with lung adenocarcinoma (LUAD). These five m6A regulatory factors exhibited potential prognostic value for the 1, 3, and 5-years survival outcomes of LUAD patients. Our findings revealed that several signaling pathways, such as cell cycle, DNA replication, RNA degradation, RNA polymerase, nucleotide excision repair and basal transcription factors, are activated in the high-risk group of LUAD patients.

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