Cell Cycle In Lung Adenocarcinoma Research Articles

Metabolic reprogramming-driven homologous recombination and TCA cycle dysregulation contribute to poor prognoses in lung adenocarcinoma.

Increasing evidence has shown that homologous recombination (HR) and metabolic reprogramming are essential for cellular homeostasis. These two processes are independent as well as closely intertwined. Nevertheless, they have rarely been reported in lung adenocarcinoma (LUAD). We analysed the genomic, immune microenvironment and metabolic microenvironment features under different HR activity states. Using cell cycle, EDU and cell invasion assays, we determined the impacts of si-SHFM1 on the LUAD cell cycle, proliferation and invasion. The levels of isocitrate dehydrogenase (IDH) and α-ketoglutarate dehydrogenase (α-KGDH) were determined by ELISA in the NC and si-SHFM1 groups of A549 cells. Finally, cell samples were used to extract metabolites for HPIC-MS/MS to analyse central carbon metabolism. We found that high HR activity was associated with a poor prognosis in LUAD, and HR was an independent prognostic factor for TCGA-LUAD patients. Moreover, LUAD samples with a high HR activity presented low immune infiltration levels, a high degree of genomic instability, a good response status to immune checkpoint blockade therapy and a high degree of drug sensitivity. The si-SHFM1 group presented a significantly higher proportion of cells in the G0/G1 phase, lower levels of DNA replication, and significantly lower levels of cell migration and both TCA enzymes. Our current results indicated that there is a strong correlation between HR and the TCA cycle in LUAD. The TCA cycle can promote SHFM1-mediated HR in LUAD, raising their activities, which can finally result in a poor prognosis and impair immunotherapeutic efficacy.

Unlocking the role of EIF5A: A potential diagnostic marker regulating the cell cycle and showing negative correlation with immune infiltration in lung adenocarcinoma

BackgroundDespite EIF5A upregulation related to tumor progression in LUAD (lung adenocarcinoma), the underlying mechanisms remain elusive. In addition, there are few comprehensive analyses of EIF5A in LUAD. MethodsWe investigated the EIF5A expression level in LUAD patients using data from the TCGA and GEO databases. We employed qRT-PCR and western blot to verify EIF5A expression in cell lines, while immunohistochemistry was utilized for clinical sample analysis. We analyzed EIF5A expression in tumor-infiltrating immune cells using the TISCH database and assessed its association with immune infiltration in LUAD using the “ESTIMATE” R package. Bioinformatics approaches were developed to discover the EIF5A-related genes and explore EIF5A potential mechanisms in LUAD. Proliferation ability was verified through CCK-8, clone formation, and EdU assays, while flow cytometry assessed apoptosis and cell cycle. Western blot was used to detect the expression of pathway-related proteins. ResultsEIF5A was significantly upregulated in LUAD. Moreover, we constructed a MAZ-hsa-miR-424-3p-EIF5A transcriptional network. We explored the potential mechanism of EIF5A in LUAD and further investigated the cAMP signaling pathway and the cell cycle. Finally, we proved that EIF5A silencing induced G1/S Cell Cycle arrest, promoted apoptosis, and inhibited proliferation via the cAMP/PKA/CREB signaling pathway. ConclusionEIF5A serves as a prognostic biomarker with a negative correlation to immune infiltrates in LUAD. It regulated the cell cycle in LUAD by inhibiting the cAMP/PKA/CREB signaling pathway.

Constructing an APOBEC-related gene signature with predictive value in the overall survival and therapeutic sensitivity in lung adenocarcinoma

BackgroundAPOBEC family play an important role in cancer mutagenesis and tumor development. The role of APOBEC family in lung adenocarcinoma (LUAD) has not been studied comprehensively. Materials and methodsThe expression data of pan-cancer as well as LUAD was obtained from public databases. The expression level of APOBEC family genes was analyzed in different normal and cancer tissues. APOBEC mutagenesis enrichment score (AMES) was utilized to evaluate the APOBEC-induced mutations and the relation of APOBEC with genomic instability. Gene set enrichment analysis was used to identify differentially enriched pathways. Univariate Cox regression and Lasso regression were applied to screen key prognostic genes. The immune cell infiltration was estimated by CIBERSORT. RT-qPCR assay, CCK-8 and Transwell assay were conducted to explore gene expression and lung cancer cell invasion. ResultsCancer tissues had significantly altered expression of APOBEC family genes and the expression patterns of APOBEC family were different in different cancer types. APOBEC3B was the most aberrantly expressed in most cancer types. In LUAD, we observed a significantly positive correlation of AMES with intratumor heterogeneity (ITH), tumor neoantigen burden (TNB), and tumor mutation burden (TMB). High AMES group had high mutation counts of DNA damage repair pathways, and high enrichment of cell cycle and DNA repair pathways. We identified four prognostic genes (LYPD3, ANLN, MUC5B, and FOSL1) based on AMES, and constructed an AMES-related gene signature. The expressions of four genes were enhanced and accelerated the invasion ability and viability of lung cancer cells. Furthermore, we found that high group increased oxidative stress level. ConclusionsAPOBEC family was associated with genomic instability, DNA damage-related pathways, and cell cycle in LUAD. The AMES-related gene signature had a great potential to indicate the prognosis and guide immunotherapy/chemotherapy for patients suffering from LUAD.

MiR-139-5p Targeting CCNB1 Modulates Proliferation, Migration, Invasion and Cell Cycle in Lung Adenocarcinoma.

Lung adenocarcinoma (LUAD) is the most frequent histological subtype of non-small cell lung cancer. Cyclin B1 (CCNB1) is the vital initiator and controller of mitosis. Studies have indicated that CCNB1 overexpression is closely associated with cell proliferation and tumorigenesis in many cancers. Thus, discovery of molecular mechanism of CCNB1 in LUAD is conducive to developing new diagnostic or therapeutic targets for LUAD. We acquired mature miRNA and mRNA expression information of LUAD from TCGA database, as well as related clinical data. CCNB1 expression in normal and LUAD tissue was analyzed. Relationship between CCNB1 and patient's survival and clinical stage was analyzed. Upstream regulatory gene miRNA of CCNB1 was predicted. qRT-PCR and western blot examined expression levels of CCNB1 and miR-139-5p in cells. CCK-8 tested cell proliferation. Scratch healing and Transwell determined cell migration and invasion. Flow cytometry analyzed the cell cycle. Dual-luciferase verified targeting relationship between the two genes. Compared to controls, CCNB1 expression was prominently high in LUAD patient samples, and associated with advanced tumor stages and shorter overall survival. MiR-139-5p expressed an evidently negative correlation with CCNB1 and was predicted to target CCNB1. MiR-139-5p mimics reduced CCNB1 mRNA and protein expression, and suppressed luciferase activity in a target-specific manner, as confirmed by a control construct with a mutated miR-139-5p binding site. CCNB1 overexpression fostered progression of LUAD cells. Mechanistically, miR-139-5p might negatively regulate CCNB1 in LUAD, thereby suppressing cell proliferation, migration, invasion and cell cycle.

Long noncoding RNA LMO7DN inhibits cell proliferation by regulating the cell cycle in lung adenocarcinoma

In our previous study, we reported that the long noncoding RNA, LMO7 downstream neighbor (LMO7DN), has a strong prognostic value in lung adenocarcinoma (LUAD). In this study, we further investigated the role of LMO7DN in LUAD progression. LMO7DN was found to be expressed at low levels in LUAD tissues, and its high expression predicted good prognosis. Bioinformatics analysis indicated that LMO7DN was closely associated with the cell cycle. Furthermore, we found that cell proliferation was significantly enhanced following knockdown of LMO7DN, and the number of cells in the G2/M phase was markedly decreased, whereas there was no change in apoptosis. Thus, LMO7DN inhibits cell proliferation by affecting the cell cycle and is of significant prognostic value in LUAD.

Non-SMC condensin I complex subunit H (NCAPH), a regulator of cell cycle, predicts poor prognosis in lung adenocarcinoma patients: a study mainly based on TCGA and GEO database.

BackgroundLung adenocarcinoma (LUAD) is the main sub-type of lung cancer, which is a major disease of human death. However, the role of non-SMC condensin I complex subunit H (NCAPH) in LUAD and its possible upstream regulation microRNAs (miRNAs) remains unclearly.MethodsIn this study, we analyzed the NCAPH mRNA and protein expression in normal and cancer tissues mainly based on Human Protein Atlas (HPA) database, The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. With the help of the Kaplan Meier plotter, we explored the prognosis role in LUAD. Furtherly, the co-expressed genes of NCAPH in LUAD were obtained by using cBioPortal, GEPIA and UALCAN database. Then, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of co-expression genes of NCAPH was conducted by DAVID, while the protein-protein interaction (PPI) network was constructed with STRING and hub genes were identified and visualized by Cytoscape software. We also investigated the miRNAs and chemicals that may downregulated the NCAPH expression.ResultsThe results showed that NCAPH expression level was elevated in LUAD tissue compared with normal lung tissue and predicted poor prognosis. GO and KEGG pathway enriched analysis of co-expressed genes suggested that NCAPH may play an important role in cell cycle in LUAD. Nine top hub co-expressed genes were all negatively related to the LUAD prognosis. Lastly, 8 miRNAs and 5 chemicals were identified to have the potential to down-regulate the NCAPH expression.ConclusionsOur study indicated that NCAPH expression in LUAD is a poor prognostic indicator, which may be the potential therapeutic target in the future.

Histone methyltransferase SETD2 inhibits tumor growth via suppressing CXCL1-mediated activation of cell cycle in lung adenocarcinoma.

The histone H3 lysine 36 methyltransferase SET-domain-containing 2 (SETD2) has been reported to be frequently mutated or deleted in many types of human cancer. However, the role of SETD2 in lung adenocarcinoma (LUAD) has not been well documented. In the present study, we found that SETD2 was significantly down-regulated both in LUAD tissues and cell lines. Functionally, the increased expression of SETD2 significantly attenuated the proliferation of cancer cells by affecting the cell cycle, whereas SETD2 deficiency dramatically improved these proliferative abilities of cancer cells. Through conjoint analysis of RNA-seq and ChIP data, we identified a functional target gene of SETD2, CXCL1, and its expression was negatively correlated with that of SETD2. Moreover, SETD2 deletion stimulated cell cycle-related proteins to promote LUAD. Further mechanistic studies demonstrated that histone H3 lysine 36 trimethylation (H3K36me3) catalyzed by SETD2 interacted with the promoter of CXCL1 to regulate its transcription and downstream signaling pathways, contributing to tumorigenesis in vitro and in vivo. Our findings suggested that SETD2 inhibited tumor growth via suppressing CXCL1-mediated activation of cell cycle, indicating that the regulation of H3K36me3 level by targeting SETD2 and/or the administration of downstream CXCL1 might represent a potential therapeutic way for new treatment in LUAD.

Sauchinone: a prospective therapeutic agent-mediated EIF4EBP1 down-regulation suppresses proliferation, invasion and migration of lung adenocarcinoma cells.

Lung adenocarcinoma (LUAD) is the top prevalent histological kind of lung cancer worldwide. Recent evidences have demonstrated that Sauchinone plays an anticancer role in tumor cell invasion and migration. Therefore, we performed this investigation to explain the potential role of Sauchinone in LUAD as well as the potential mechanism involved. Cell counting kit 8 (CCK-8) and transwell experiments were implemented to measure the proliferative, invasive and migratory abilities of LUAD cells. qRT-PCR and Western blot were performed to detect the transfection efficiency of si-EIF4EBP1s. Additionally, Western blot was also implemented to evaluate the effect of Sauchinone on EIF4EBP1 expression level as well as cell cycle-related proteins. Our findings showed that Sauchinone remarkably suppressed the proliferative ability of LUAD cells in a dose-dependent and time-dependent manner. EIF4EBP1 was a candidate target gene of Sauchinone. EIF4EBP1 expression was increased in LUAD tissues, and its high expression induced a poorer prognosis of LUAD patients. EIF4EBP1 expression was positively associated with cell cycle in LUAD. Sauchinone treatment attenuated EIF4EBP1 expression and cell cycle-related protein levels. Knockdown of EIF4EBP1 repressed the proliferation, invasion and migration of LUAD cells; furthermore, Sauchinone stimulation enforced its inhibitory effect. Meanwhile, the treatment of Sauchinone intensified the arrest of cell cycle induced by EIF4EBP1 knockdown. To sum up, our discovery indicated that Sauchinone exerts an anticancer role through down-regulating EIF4EBP1 and mediating cell cycle in LUAD.

UHRF1 predicts poor prognosis by triggering cell cycle in lung adenocarcinoma.

Accumulating evidence suggests that ubiquitin‐like with plant homeodomain and ring finger domains 1 (UHRF1) is overexpressed in non‐small cell lung cancer (NSCLC); however, the expression and function of UHRF1 in the subtype of NSCLC are still unclear. Here, we investigate the expression and prognosis traits of UHRF1 in large NSCLC cohorts and explore the molecular characters during UHRF1 up‐regulation. We find that UHRF1 is predominantly overexpressed in lung squamous cell carcinoma (SCC). Surprisingly, the up‐regulated UHRF1 is only associated with the overall survival of lung adenocarcinoma (ADC) and knockdown of UHRF1 dramatically attenuates ADC tumorigenesis. Mechanically, we identify a hub gene that includes a total of 55 UHRF1‐related genes, which are tightly associated with cell cycle pathway and yield to the poor clinical outcome in ADC patients. What's more, we observe knockdown of UHRF1 only affects ADC cells cycle and induces cell apoptosis. These results suggest that up‐regulated UHRF1 only contributes to lung ADC survival by triggering cell cycle pathway, and it may be a prognostic biomarker for lung ADC patients.

Long Non-Coding RNAs Suppressed Cell Cycle in Lung Adenocarcinoma

BACKGROUND/AIMS: Lung cancer is the leading cause of cancer-associated mortality worldwide. Long noncoding RNAs (lncRNAs) have been found to be related to different cancers and play important roles in regulating the pathological and physiological processes of numerous cancers. The aim of the current study was to identify potential prognostic long non-coding RNA biomarkers for predicting survival in patients with lung adenocarcinoma (LUAD) using The Cancer Genome Atlas (TCGA) dataset. METHODS: The clinical pathology and gene expression profiles of lung cancer were downloaded from The Cancer Genome Atlas. The differentially expressed lncRNAs were identified and patient survival analysis performed. The prognostic signature was further evaluated by functional assessment and bioinformatics analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to predict the function of lncRNAs and to construct a gene co-expression network relationship to predict downstream target genes with the aid of the STRING database and Cytoscape software RESULTS: lncRNA RP11-434D9.1 and FENDRR had lower expression levels in lung cancer and were associated with poor prognoses. We found that the major functions of lncRNA RP1-434D9.1 and FENDRR were related to the cell cycle, and we further screened out the possible downstream target genes CDK1, CCNB1, CCNB2, CCNA1, AURKA, PLK1. CONCLUSIONS: LncRNA RP11-434D9.1 and FENDRR can be used as molecular biomarkers for the diagnosis and prognosis of lung adenocarcinoma. These dysregulated lncRNAs might negatively regulate cell cycle through the hub genes, which then affected cancer cell proliferation. These results may provide new ideas for further clinical research for markers used for lung cancer diagnosis and prognosis. FUNDING STATEMENT: This work was supported by National Natural Science Foundation of China (81672270); Key project of Guangzhou Science Technology and Innovation Committee (201707020042); Science and Technology Program of Guangzhou (201803010024); Science and Technology Planning Project of Guangdong Province, China (2017B020226005); Science and Technology Program of Guangzhou (201903010003). Clinical Research Startup Program of Southern Medical University by High-level University Construction Funding of Guangdong Provincial Department of Education (LC2016PY013); Scientific Research Projects of Wu Jieping Medical Research Foundation (320.6750.17206). DECLARATION OF INTERESTS: The authors declare that no conflicts of interest exist. ETHICS APPROVAL STATEMENT: Every specimen was anonymously handled based on ethical standards. All participants provided signed informed consent and our study had full approval from the hospital’s Ethical Review Committee.

TEAD4 exerts pro-metastatic effects and is negatively regulated by miR6839-3p in lung adenocarcinoma progression.

Several studies have shown the tumorigenesis role of transcriptional enhancer associate domain (TEAD) proteins; here, we initially explored expression, function and signalling mechanisms of TEAD4 in lung adenocarcinoma (LAD). Both the mRNA and protein levels of TEAD4 were increased in LAD tissues than those in adjacent nontumourous tissues. Besides, database search indicated a poorer clinical outcome in LAD patients with higher TEAD4 expression, revealing its potential tumour‐promoting role. Therefore, we conducted cellular experiments to further investigate its effect on tumour phenotypes. Accordingly, TEAD4 showed little impact on LAD cell cycle, proliferation, or apoptosis. However, silencing TEAD4 remarkably attenuated cell migration and invasion capacities. Consistently, several important epithelial‐mesenchymal transition (EMT) markers such as E‐cadherin and Slug were consequently altered by silencing TEAD4. Furthermore, we identified a novel TEAD4‐targeted microRNA, namely miR6839‐3p, and confirmed its function in suppressing TEAD4 expression. Finally, the impact of overexpressing miR6839‐3p mimics on LAD progression was validated, which showed a similar pattern with TEAD4 knockdown cells. Taken together, our data not only revealed the significant role of TEAD4 in promoting LAD progression and predicting clinical outcome but also distinguished miR6839‐3p mimics as a promising therapeutic direction.

Abstract A45: High CDC2_1 activatory functions and mechanisms in lung adenocarcinoma

Abstract In order to study high CDC2_1 feedback-interactive cell cycle mechanism in lung adenocarcinoma, high CDC2_1 feedback-interactive multidimensional molecular network was constructed from mutual positive correlation of the overlapping with CDC2_1 direct activatory relationship by GRNInfer and Pearson correlation coefficient CC ≥0.25 in lung adenocarcinoma (GSE7670 from Taiwan samples). Our result showed a loop from TOP2A_2 to CKS1B to RRM2_1, respectively; There are interactions among TOP2A_2 to CKS1B to RRM2_1, respectively. Therefore, we proposed high CDC2_1 feedback-interactive TOP2A_2 interactive CKS1B interactive RRM2_1 loop. Based on GO, KEGG, GenMAPP, BioCarta and disease databases, RRM2_1, TOP2A_2, CKS1B knowledge included DNA replication, cytoplasm, protein-binding, nucleoplasm, cell cycle, cell division, etc. We further put forward high CDC2_1 feedback-interactive cell cycle in lung adenocarcinoma. In order to study high CDC2_1 activated-upstream mitosis mechanism in lung adenocarcinoma, high CDC2_1 activated-upstream multidimensional molecular network was constructed. Our result showed interactions among NCAPG_1 to SPAG5 to NDC80, respectively. Therefore, we proposed high CDC2_1 activated-upstream NCAPG_1 to SPAG5 to NDC80 interaction. Based on GO, KEGG, GenMAPP, BioCarta and disease databases, NCAPG_1, SPAG5, NDC80 knowledge included M phase, nucleus, cytoplasm, protein-binding, mitosis, condensed chromosome kinetochore, spindle organization and biogenesis, phosphoinositide-mediated signaling, condensin complex, mitotic chromosome condensation, spindle microtubule, cytokinesis, chromosome, pericentric region, mitotic sister chromatid segregation, etc. We further put forward high CDC2_1 activated-upstream mitosis in lung adenocarcinoma. In order to study high CDC2_1 activating-downstream mitosis mechanism in lung adenocarcinoma, high CDC2_1 activating-downstream multidimensional molecular network was constructed. Our result showed mutual interaction among EPN3, NUSAP1_1, PRC1. Therefore, we proposed high CDC2_1 activating-downstream EPN3/NUSAP1_1/PRC1 complex. Based on GO, KEGG, GenMAPP, BioCarta and disease databases, NCAPG_1, SPAG5, NDC80 knowledge included nucleus, cytoplasm, lipid-binding, cell cycle, cell division, cytokinesis, cytoplasmic vesicle, microtubule, DNA-binding, cytokinesis after mitosis, mitotic chromosome condensation, establishment of mitotic spindle localization, protein-binding, mitotic spindle elongation, Embryonic Stem Cell, microtubule cytoskeleton, etc. We further put forward high CDC2_1 activating-downstream mitosis in lung adenocarcinoma. High CDC2_1 feedback-interactive, activated-upstream and activating-downstream molecular networks were reverse-verified by the corresponding high CDC2_1 repressive network in lung adenocarcinoma, activatory and repressive in human normal adjacent lung adenocarcinoma tissues, respectively. Citation Format: Lin Wang, Juxiang Huang, Minghu Jiang, Qingchun Chen, Zhenfu Jiang, Haitao Feng. High CDC2_1 activatory functions and mechanisms in lung adenocarcinoma. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr A45.