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.

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.

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 Aberrant Long Noncoding RNAs Expression Profiles Affect Cisplatin Resistance in Lung Adenocarcinoma

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Lijuan Hu ◽  
Jian Chen ◽  
Fan Zhang ◽  
Junjun Wang ◽  
Jingye Pan ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
A549 Cell ◽  
Cisplatin Resistance ◽  
Expression Profiles ◽  
Noncoding Rnas ◽  
Long Noncoding Rnas ◽  
Pcr Analysis ◽  
Sensitive Cell ◽  
Chain Reaction ◽  
Polymerase Chain

Background. Long noncoding RNAs (lncRNAs) have been shown to be involved in the mechanism of cisplatin resistance in lung adenocarcinoma (LAD). However, the roles of lncRNAs in cisplatin resistance in LAD are not well understood. Methods. We used a high-throughput microarray to compare the lncRNA and mRNA expression profiles in cisplatin resistance cell A549/DDP and cisplatin sensitive cell A549. Several candidate cisplatin resistance-associated lncRNAs were verified by real-time quantitative reverse transcription polymerase chain reaction (PCR) analysis. Results. We found that 1,543 lncRNAs and 1,713 mRNAs were differentially expressed in A549/DDP cell and A549 cell, hinting that many lncRNAs were irregular from cisplatin resistance in LAD. We also obtain the fact that 12 lncRNAs were aberrantly expressed in A549/DDP cell compared with A549 cell by quantitative PCR. Among these, UCA1 was the aberrantly expressed lncRNA and can significantly reduce the IC50 of cisplatin in A549/DDP cell after knockdown, while it can increase the IC50 of cisplatin after UCA1 was overexpressed in NCI-H1299. Conclusions. We obtained patterns of irregular lncRNAs and they may play a key role in cisplatin resistance of LAD.

scholarly journals Aberrant Expression of SCARA5 in Lung Adenocarcinoma and its Clinical Significance

2020 ◽  
Author(s):  
Genhua Yu ◽  
Xufeng Gong ◽  
Aixia Fu ◽  
Yan Hou ◽  
Jingping Xia ◽  
...  
Keyword(s):  
Overall Survival ◽  
Lung Adenocarcinoma ◽  
A549 Cell ◽  
Quantitative Polymerase Chain Reaction ◽  
Scavenger Receptor ◽  
Prognostic Significance ◽  
Area Under The Curve ◽  
Suppressor Gene ◽  
Aberrant Expression

Abstract BackgroundLung adenocarcinoma, the main subtype of non-small cell lung cancer, leading one of the most aggressive and fatal causes of malignant deaths around the world. Scavenger receptor class A member 5 (SCARA5), a newly discovered tumor suppressor gene, belonged to the SR family. The present study’s object was to explore the clinical impacts of SCARA5 in lung adenocarcinoma treatment. MethodsSCARA5 expressions in 120 paired lung adenocarcinoma patients’ tissues and cell lines were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC). CCK-8, EdU, flow cytometry, and western blot assays further demonstrated the significance of SCARA5 on A549 cell growth. Then, the relationships between the expression level of SCARA5 and pathological factors were analyzed. Finally, the receiver operating characteristic (ROC) curve and overall survival analysis was carried out to verify the diagnostic and prognostic significance of SCARA5 in lung adenocarcinoma. ResultsSCARA5 was prominently decreased in lung adenocarcinoma cells and tissues compared with human bronchial epithelial cells and para-carcinoma non-tumor tissues. Meanwhile, SCARA5 expression was strongly correlated with smoking (P = 0.0011), TNM stage (P < 0.0001) and lymph node metastasis (P = 0.0005). Furthermore, SCARA5 may be a crucial biomarker for lung adenocarcinoma diagnosis with an area under the curve (AUC) of 0.9102 while SCARA5 could significantly reduce overall survival (OS; P = 0.0006). In vitro experiments, we found that SCARA5 overexpressing could significantly hinder A549 cell proliferation but facilitate apoptosis through the AKT signaling pathway. ConclusionsSCARA5 might be an important diagnostic and prognostic biomarker for patients with lung adenocarcinoma.

scholarly journals Characterization of ceRNA network to reveal potential prognostic biomarkers in triple-negative breast cancer

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7522 ◽  
Author(s):  
Xiang Song ◽  
Chao Zhang ◽  
Zhaoyun Liu ◽  
Qi Liu ◽  
Kewen He ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Quantitative Polymerase Chain Reaction ◽  
The Cancer Genome Atlas ◽  
Expression Level ◽  
Cerna Network ◽  
Kaplan Meier ◽  
Cancer Genome Atlas ◽  
Non Coding Rnas

Triple-negative breast cancer (TNBC) is a particular subtype of breast malignant tumor with poorer prognosis than other molecular subtypes. Previous studies have demonstrated that some abnormal expression of non-coding RNAs including microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) were closely related to tumor cell proliferation, apoptosis, invasion, migration and drug sensitivity. However, the role of non-coding RNAs in the pathogenesis of TNBC is still unclear. In order to characterize the molecular mechanism of non-coding RNAs in TNBC, we downloaded RNA data and miRNA data from the cancer genome atlas database. We successfully identified 686 message RNAs (mRNAs), 26 miRNAs and 50 lncRNAs as key molecules for high risk of TNBC. Then, we hypothesized that the lncRNA–miRNA–mRNA regulatory axis positively correlates with TNBC and constructed a competitive endogenous RNA (ceRNA) network of TNBC. Our series of analyses has shown that five molecules (TERT, TRIML2, PHBP4, mir-1-3p, mir-133a-3p) were significantly associated with the prognosis of TNBC, and there is a prognostic ceRNA sub-network between those molecules. We mapped the Kaplan–Meier curve of RNA on the sub-network and also suggested that the expression level of the selected RNA is related to the survival rate of breast cancer. Reverse transcription-quantitative polymerase chain reaction showed that the expression level of TRIML2 in TNBC cells was higher than normal. In general, our findings have implications for predicting metastasis, predicting prognosis and discovering new therapeutic targets for TNBC.

scholarly journals In vivo and in vitro effects of hyperplasia suppressor gene on the proliferation and apoptosis of lung adenocarcinoma A549 cells

2018 ◽  
Vol 38 (5) ◽  
Author(s):  
Zhen-Qing Sun ◽  
Gang Chen ◽  
Qiang Guo ◽  
He-Fei Li ◽  
Zhou Wang
Keyword(s):  
Cell Proliferation ◽  
Lung Adenocarcinoma ◽  
Gene Silencing ◽  
Nude Mice ◽  
A549 Cell ◽  
A549 Cells ◽  
Suppressor Gene ◽  
Proliferation And Apoptosis ◽  
Lung Adenocarcinoma A549

Lung adenocarcinoma is the most common subtype of non-small cell lung cancer (NSCLC). Hyperplasia suppressor gene (HSG) has been reported to inhibit cell proliferation, migration, and remodeling in cardiovascular diseases. However, there lacks systematic researches on the effect of HSG on the apoptosis and proliferation of lung adenocarcinoma A549 cells and data of in vivo experiments. The present study aims to investigate the effects of HSG gene silencing on proliferation and apoptosis of lung adenocarcinoma A549 cells. The human lung adenocarcinoma A549 cell was selected to construct adenovirus vector. Reverse transcription-quantitative PCR (RT-qPCR) and Western blot analysis were conducted to detect expressions of HSG and apoptosis related-proteins. Cell Counting Kit (CCK)-8 assay was performed to assess A549 cell proliferation and flow cytometry to analyze cell cycle and apoptosis rate. The BALB/C nude mice were collected to establish xenograft model. Silenced HSG showed decreased mRNA and protein expressions of HSG, and elevated A549 cell survival rates at the time point of 24, 48, and 72 h. The ratio of cells at G0/G1 phase and apoptosis rate decreased and the ratio of cells at S- and G2/M phases increased following the silencing of HSG. There were decreases of B cell lymphoma-2 (Bcl-2)-associated X protein (Bax), Caspase-3, and Caspase-8 expressions but increases in Bcl-2 induced by silenced HSG. As for the xenograft in nude mice, tumor volume increased, and apoptosis index (AI) decreased after HSG silencing. These results indicate that HSG gene silencing may promote the proliferation of A549 cells and inhibit the apoptosis. HSG may be a promising target for the treatment of lung adenocarcinoma.

scholarly journals NRAS Expression is Associated With Prognosis and Tumor Immune Microenvironment in Lung Adenocarcinoma

2021 ◽  
Author(s):  
Yueren Yan ◽  
Zhendong Gao ◽  
Han Han ◽  
Yue Zhao ◽  
Yang Zhang ◽  
...  
Keyword(s):  
T Cells ◽  
Lung Adenocarcinoma ◽  
Enrichment Analysis ◽  
Gene Set Enrichment Analysis ◽  
The Cancer Genome Atlas ◽  
Expression Level ◽  
Immune Microenvironment ◽  
Kaplan Meier ◽  
Tumor Immune Microenvironment ◽  
Cox Analysis

Abstract Purpose: NRAS plays a pivotal role in progression of various kinds of somatic malignancies; however, the correlation between NRAS and lung adenocarcinoma is less known. We aim to analyze the prognostic value of NRAS expression in lung adenocarcinoma, and explore the relationship between NRAS and tumor immune microenvironment. Methods: We obtained the transcriptome pofiles and clinical data of LUAD from The Cancer Genome Atlas database and three Genome Expression Omnibus datasets. Specimens from 325 patients with completely resected lung adenocarcinoma were collected for immunohistochemical assays of NRAS, PD-L1, PD-1 and TIM-3. Then we performed gene set enrichment analysis to investigate cancer-related and immune-related signaling pathways. TIMER algorithms were performed to evaluate tumor immune infiltrating cells and immune-related biomarkers.Results: Compared with adjacent non-tumor tissue, NRAS expression was significantly upregulated in LUAD tissue. NRAS expression was significantly correlated with more advanced stage and positive lymph nodes. Kaplan-Meier curves and Cox analysis suggested that high NRAS expression led to a poor prognosis, and could be an independent prognostic factor in LUAD patients. Besides, NRAS expression was positively correlated with CD8+ T cells, macrophages, and neutrophils, and negatively correlated with B cells and CD4+ T cells. The expression level of NRAS was positively correlated with PD-L1, PD-1, and TIM-3 both at RNA and protein level. Conclusions: To conclude, we found NRAS a novel prognostic biomarker in LUAD. Besides, the expression level of NRAS may influence the prognosis of LUAD via various kinds of cancer-related pathways and remodeling TIM.

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.

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