Baoen Shan

Increasing evidence showed that the substance P (SP)/neurokinin‑1 receptor (NK1R) complex is involved in the development of several cancers. However, little is known about the mechanisms by which SP/NK1R complex plays a role in esophageal squamous cell carcinoma (ESCC) progression. RT‑qPCR, CCK‑8, Transwell, western blotting, immunohistochemical, immunofluorescence, ELISA and analysis of apoptosis were employed in the present study. It was aimed to investigate the function and therapeutic potential of the SP/tr‑NK1R system in human ESCC progression. The results revealed that both SP and tr‑NK1R were highly expressed in ESCC cell lines and specimens. In ESCC tissues, SP was mainly derived from ESCC cells and M2 macrophages. The NK1R antagonist aprepitant inhibited the SP‑induced proliferation of human ESCC cell lines. Aprepitant inhibited cell migration and invasion and induced apoptosis of ESCC cells by downregulating the PI3K/AKT/mTOR signaling pathways. Animal experiments revealed that aprepitant inhibited tumor progression of ESCC in xenograft mice. In conclusion, high expression of SP plus tr‑NK1R indicated poor prognosis in ESCC, suggesting that aprepitant has a potential application in ESCC. To the best of our knowledge, high SP and tr‑NK1R expression in ESCC cell lines was reported for the first time in the present study. These findings provided evidence for a novel therapeutic strategy for patients with ESCC.

Context . Although great progress has occurred in breast cancer (BC) treatment, including chemotherapy, chemoradiotherapy, and surgical resection, the rate of patients' survival is still unsatisfactory. Multiple genes and factors have proven to contribute to BC's occurrence. Thus, it's urgent to explore the molecular mechanisms in the development and progression of BC and find possible targets for therapy. Objective . The study intended to assess the mechanisms of miR-518a-5p's effect on BC by targeting the zinc finger E-box-binding homeobox 2 (ZEB2) gene. Design . The research team designed a laboratory study and retrospective analysis. Setting . The study took place in the Department of Breast Surgery at the Xingtai People's Hospital in Xingtai, Hebei, China. Outcome Measures . The study measured the miR-518a-5p expression in BC tissues and normal tissues using a real-time quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) test. The research team purchased the BC cells MDA-MB-231 and MCF-7 and measured the effects of the miR-518a-5p on BC cell activity as well as epithelial-mesenchymal transition (EMT) ability, using cell scratch tests and Transwell assays. The team assessed the ZEB2 protein expression and verified the targeting relationship using a Dual-Luciferase Reporter assay. Results . The miR-518a-5p expressed was low in the BC tissues and cell lines compared with adjacent normal tissues (P <.05). Overexpressing the miR-518a-5p inhibited BC-cell migration and invasion (P <.05). Moreover, the ZEB2 was the target gene for the miR-518a-5p. The Luciferase assay verified that the miR-518a-5p directly targeted the ZEB2 in BC cells (P <.05). The Transwell invasion assay, western blot analysis, and wound healing assay also showed that the miR-518a-5p inhibited BC cells by targeting ZEB2 (P <.05). Conclusions . The miR-518a-5p suppressed BC migration, invasion, and process of EMT by regulating ZEB2. In the future, this method may be a new option for BC diagnosis and treatment. An in-depth understanding of the role of the miR-518a-5p in BC can help clinicians to understand the pathogenic mechanism of breast cancer more accurately.

Background: Esophageal squamous cell carcinoma (ESCC) is a common gastrointestinal malignant tumor, while the molecular mechanisms have not been fully elucidated. Multiple circular RNAs have been reported to involve in the onset and progression of malignant tumors through various molecular mechanisms. However, the clinical significance and functional mechanism of most circRNAs involved in the progression of ESCC remains obscure. Methods: RNA-Seq was used to explore potential circRNAs in participated in 5 pairs of ESCC and their corresponding normal esophageal tissues. The up-regulated circCYP24A1 was selected. Fluorescence in situ hybridization was cunducted to verificated the expression and intracellular localization of circCYP24A1 by using the tissue microarray. The Kaplan–Meier method and Cox proportional hazards model was used to examine the potential prognostic value of circCYP24A1 on overall survival of ESCC patients. The biological function were confirmed by gain- and loss-of-function approaches in vivo. mRNA expression profile microarray was proformed to investigate the downstream signaling pathways involved in circCYP24A1. RNA pull-down assay and mass spectrometry were performed to identify the proteins associated with circCYP24A1. Rescue experiments were carried out to identified hypothetical regulatory role of circCYP24A1 on ESCC progression in vivo and in virto. Results: In this study, we identified circCYP24A1 in ESCC tissues by RNA sequencing, which is up-regulated in 114 cases of ESCC tissues and acts as a novel prognosis-related factor. Moreover, circCYP24A1 promoted the ability of proliferation, migration, invasion and clone formation in vitro, as well as tumor growth in vivo. Mechanistically, chemokine (C-Cmotif) ligand 5 (CCL5) is functional downstream mediator for circCYP24A1, which is screened by mRNA microarray. Moreover, circCYP24A1 physically interacts with M2 isoform of pyruvate kinase (PKM2). Rescue experiments showed that PKM2 knockdown partly reverses the promotional effects of circCYP24A1. It was revealed that circCYP24A1 increases secretion of CCL5 through the mechanism mainly by interacting with PKM2, an activator of NF-κB pathway, and thereby accelerate malignant progression of ESCC. Conclusions: Up-regulated circCYP24A1 could activate NF-κB pathway by binding PKM2, which promotes the secretion of CCL5 and accelerate malignant progression of ESCC. Our fndings recommended a novel function for circCYP24A1 as a potential effective biomarker for judging prognosis and a therapeutic target in ESCC.

Circular RNAs (circRNAs) are a type of noncoding RNA, which play a vital role in the occurrence and development of esophageal squamous cell carcinoma (ESCC). While the role of novel circADAMTS6 in ESCC remains unknown. We assessed circADAMTS6 expression in ESCC tissues and cells, and the relationship between circADAMTS6 expression and overall survival of ESCC patients. Functional experiments in vitro and xenograft in vivo assay were applied to explore the functions and mechanisms of circADAMTS6 in ESCC. Results found that up-regulation of circADAMTS6 was associated with poor overall survival and may acted as an independent risk factor for ESCC prognosis. Knockdown of circADAMTS6 significantly inhibited the proliferation, migration and invasion of ESCC cells and growth of xenograft tumors in vivo. Induced AGR2 expression was able to rescue the loss of function induced by si-circADAMTS6 in KYSE150 cell. CircADAMTS6 may acts as oncogene by activating AGR2 and the Hippo signaling pathway coactivator YAP in ESCC.

Background: Circular RNAs (circRNAs) are involved in the tumorigenesis and progression of lung adenocarcinoma (LUAD). This study aimed to determine the role of circ_0072088 in LUAD.Methods: The existence and expression of circ_0072088 in human LUAD tissues and cell lines were determined through Sanger sequencing, quantitative reverse transcription‐polymerase chain reaction, and fluorescence in situ hybridization (FISH). Subsequently, the biological role of circ_0072088 was examined using loss‐of‐function assays in H1299 cells. Moreover, circ_0072088/miR‐1261/PIK3CA pathway‐mediated biological effects in H1299 were verified using bioinformatic prediction and experiments, including interaction analysis (FISH, luciferase reporter, and RNA‐pulldown assays), and tumor biological function test (CCK8 and colony formation, wound healing, and transwell assays). Finally, miR‐1261 and PIK3CA expression and LUAD patient survival were further analyzed using FISH, immunohistochemical staining, and the Kaplan–Meier plotter database, respectively.Results: First, an increase in circ_0072088 was confirmed in human LUAD tissues. Thereafter, it was mainly localized in the cytoplasm and was found to enhance cell proliferation, migration, and invasion of H1299 cells. Mechanistically, circ_0072088 directly downregulated miR‐1261 expression, whereas increased PIK3CA gene expression was associated with poor overall survival of LUAD patients. The activation of the circ_0072088/miR‐1261/PIK3CA regulatory pathway may play a significant role in the tumorigenesis and progression of LUAD.Conclusions: Circ_0072088‐dependent regulation of miR‐1261/PIK3CA is important for cell proliferation, migration, and invasion during the tumorigenesis and progression of LUAD, warranting the need to consider the circ_0072088/miR‐1261/PIK3CA regulatory pathway as a potential therapeutic target in patients with lung adenocarcinoma.

Non‑small cell lung cancer (NSCLC) is one of the leading causes of cancer‑associated mortality. Lung adenocarcinoma (LAC) is the most prevalent pathological subtype of NSCLC and accounts for ~40% of all lung cancer mortalities. There remains an urgent demand for the identification of novel biomarkers for the diagnosis and development of therapeutic strategies for LAC. In the present study, the profiles of the differentially‑expressed circular RNAs (circRNAs) in LAC tissues compared with those in their corresponding non‑cancerous tissues were obtained after analyzing the circRNA microarray dataset GSE101586. The expression pattern of the indicated circRNAs in the LAC tissues were subsequently verified using reverse transcription‑quantitative PCR (RT‑qPCR). The potential prognostic significance of these circRNAs in patients with LAC were then analyzed in a retrospective clinical study. A circRNA‑microRNA (miR or miRNA)‑mRNA regulatory network in LAC was established by using Cytoscape. In addition, a protein‑protein interaction (PPI) network was plotted using the Search Tool for the Retrieval of Interacting Genes/Proteins and visualized through Cytoscape. The prognostic value of the hub genes found was then analyzed based on the Gene Expression Profiling Interactive Analysis database. In total, four differentially‑expressed circRNAs were obtained from the GSE101586 microarray dataset, three of which (hsa_circ_0006220, hsa_circ_0072088 and hsa_circ_0001666) were confirmed by RT‑qPCR to be highly expressed in LAC tissues. This retrospective clinical study revealed that higher expression levels of these three circRNAs were associated with poorer prognoses in patients with LAC. In addition, siRNA‑mediated knockdown of these circRNAs was found to inhibit cell proliferation, migration and invasion in LAC cells. Following analysis of the molecular mechanism underlying these circRNAs, eight miRNAs, namely miR‑520f, miR‑1261, miR‑1270, miR‑620, miR‑188‑3p, miR‑516b, miR‑940 and miR‑661, were identified with potential binding sites for these three circRNAs. Subsequently, 232 overlapped genes from the 795 upregulated genes in the LAC samples from The Cancer Genome Atlas database and 7,829 predicted target genes of the list of eight aforementioned miRNAs were obtained. A circRNA‑miRNA‑mRNA network was then constructed. A PPI network was established, with six hub genes, namely kinesin family member (KIF) 2C, KIF18B, maternal embryonic leucine zipper kinase, baculoviral IAP repeat‑containing 5, polo‑like kinase 1 and cytoskeleton‑associated protein 2‑like, determined from this network. Higher expression levels of each of these hub genes were found to be associated with poorer prognoses of patients with LAC. To conclude, data from the present study suggested that circRNAs hsa_circ_0006220, hsa_circ_0072088 and hsa_circ_0001666 have the potential to be viable biomarkers and therapeutic targets for LAC.