BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

In order to explore the mechanism by which Rehmannia extract enhances ovarian follicle development,reduces ovarian aging,and improves the production performance of chickens,35-week-old black skin and red crest chickens were randomly divided into a blank control group and groups supplemented with 0.1％,0.2％and 0.4％Rehmannia extract for feeding experiments.Changes of histological structure and molecular regulation for improving the reproductive perform-ance of black skin and red crest chicken were explored here by detecting the reproductive hormones and the corresponding receptors in ovary and quantifying the cell proliferation and apoptotic lesion.Based on the PI3K/AKT/mTOR and AMPK signaling pathways,mechanisms of Rehmannia ex-tract on promoting ovarian follicle development and alleviating ovarian aging were further studied to enrich the reproductive regulation theory of native breed chickens and improve their reproduc-tive performance.The results indicated that 0.4％Rehmannia extract could improve the levels of LH,E2 and P4 in serum,elevate the relative expression levels of FSHR,ERβ and PCNA mRNA in ovarian tissues,increase the content of PCNA protein,and decrease the number of apoptosis cells and the level of apoptosis protein Cleaved-Caspase-3.By enhancing the secretion of norepinephrine and activating the a2A-AR and the downstream PI3K/AKT/mTOR signaling pathway in ovary,the Rehmannia extract could effectively relieve the decline of ovarian function and maintain stable and efficient ovarian functions,which was independent of AMPK signaling pathway.Therefore,0.4％Rehmannia extract is conducive to enhancing the reproductive function via regulating PI3K/AKT/mTOR signaling pathway.

In order to explore the mechanism by which Rehmannia extract enhances ovarian follicle development,reduces ovarian aging,and improves the production performance of chickens,35-week-old black skin and red crest chickens were randomly divided into a blank control group and groups supplemented with 0.1％,0.2％and 0.4％Rehmannia extract for feeding experiments.Changes of histological structure and molecular regulation for improving the reproductive perform-ance of black skin and red crest chicken were explored here by detecting the reproductive hormones and the corresponding receptors in ovary and quantifying the cell proliferation and apoptotic lesion.Based on the PI3K/AKT/mTOR and AMPK signaling pathways,mechanisms of Rehmannia ex-tract on promoting ovarian follicle development and alleviating ovarian aging were further studied to enrich the reproductive regulation theory of native breed chickens and improve their reproduc-tive performance.The results indicated that 0.4％Rehmannia extract could improve the levels of LH,E2 and P4 in serum,elevate the relative expression levels of FSHR,ERβ and PCNA mRNA in ovarian tissues,increase the content of PCNA protein,and decrease the number of apoptosis cells and the level of apoptosis protein Cleaved-Caspase-3.By enhancing the secretion of norepinephrine and activating the a2A-AR and the downstream PI3K/AKT/mTOR signaling pathway in ovary,the Rehmannia extract could effectively relieve the decline of ovarian function and maintain stable and efficient ovarian functions,which was independent of AMPK signaling pathway.Therefore,0.4％Rehmannia extract is conducive to enhancing the reproductive function via regulating PI3K/AKT/mTOR signaling pathway.

Objective:To analyze the influencing factors of abnormal 15-minute retention rate of indocyanine green (ICG R15) (≥10%) in patients with hepatocellular carcinoma, and to construct a nomogram model, and to evaluate the prediction efficiency of the nomogram model.Methods:The clinical data of 190 patients with hepatocellular carcinoma in Zhengzhou University People's Hospital from December 2017 to June 2022 were retrospectively analyzed, including 148 males and 42 females, aged (57.8±9.9) years. According to ICG R15, the patients were divided into ICG R15 normal group ( n=134, ICG R15<10%) and ICG R15 abnormal group ( n=56, ICG R15≥10%). Univariate and multivariate logistic regression were used to analyze the influencing factors of abnormal ICG R15, and the nomogram model was established. The predictive ability of the model was evaluated by receiver operating characteristic (ROC) curve and C-index, and the model was verified by calibration curve and decision analysis curve. Results:Abnormal ICG R15 group the proportion of liver cirrhosis, albumin ≤35 g/L, hemoglobin ≤110 g/L, platelet count ≤100×10 9/L, prothrombin time >13 s, alanine aminotransferase >40 U/L, aspartate aminotransferase >40 U/L, total bilirubin >34.2 μmol/L, and the largest tumor diameter >5.0 cm, spleen volume >383.1 cm 3, spleen volume to of non-tumor liver volume (SNLR) >0.276 and liver tumor volume >117.2 cm 3 were higher than that of ICG R15 normal group, and the differences were statistically significant (all P<0.05). Logistic regression analysis showed that liver cirrhosis ( OR=3.89, 95% CI: 1.28-11.80, P=0.016), spleen volume >383.1 cm 3( OR=5.17, 95% CI: 1.38-19.38, P=0.015), SNLR >0.276 ( OR=5.54, 95% CI: 1.44-21.26, P=0.013) and total bilirubin >34.2 μmol/L( OR=10.20, 95% CI: 1.88-55.39, P=0.007) increased the risk of abnormal ICG R15. A nomogram model was constructed based on the above risk factors. The C-index of the model was 0.915 (95% CI: 0.872-0.957), and the area under the ROC curve predicted by the nomogram model was 0.915 (95% CI: 0.871-0.958). The calibration curve showed that the correlation index of the abnormal ICG R15 predicted by the nomogram was similar to actual situation. Decision analysis curve showed high returns. Conclusion:Liver cirrhosis, spleen volume >383.1 cm 3, SNLR>0.276 and total bilirubin >34.2 μmol/L were indepentlent risk factors for abnormal ICG R15 in patients with hepatocellur carcinoma. The clinical prediction model of ICG R15 abnormality constructed by nomogram has good prediction efficiency, which can provide a reference for evaluating preoperative liver reserve function of patients with hepatocellular carcinoma.

Objective To construct a Luciferace reporter vector containing the 3'untranslated region (3'UTR) of NFAT5 and measure the correlation between NFAT5 and miR-155.Methods The miR-155 targeting NFAT5 3'UTR was predicted by Target Scan,Mir Base and Pic Tar.NFAT5 and mutant NFAT5 sequence(NFAT5-mu) were then designed and synthesized,and they were cloned into pMIR-REPORTTM Luciferace reporter vector.Human embryonic kidney-293AD (HEK-293AD) cells of the 4th passage were divided into 4 groups according to the random number table.cells in plasimd +miR-155 mimics groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 mutated groups were transfected with pMIR-NFAT5-mu recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 control groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 Negative control;cells in plasimd +miR-155 inhibitor were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 inhibitor;and were respectively transfected into together by liposome.After culture for 24 h,the luciferase activity was detected by dual luciferase reporter assay system.Results TargetScan,Miranda and PicTar shared the results that NFAT5 has the complementary binding sites with 3'UTR of miR-155.And luciferase reporter vectorwas constructed.Therefore the result of sequencing and double digesting of recombined plasmid were completely correct.Dual-luciferase reporter assay showed that miR-155 possesses a target effect on 3'UTR of NFAT5.Compared to the pMIR-NFAT5 + miR-control group,the luciferase activity of the pMIR-NFAT5 + miR-1 5 5 mimics group was decreased,with statistically significant difference(P＜0.01),while there was no significant difference at other time points(P＞0.05).Conclusion The pMIR-NFAT5 recombinant plasmid and pMIR-NFAT5 recombinant mutated plasmid were confirmed with successful construction.and it was found that miR-155 can target NFAT5 mRNA 3'-UTR.The results provide the experiment data for further disclosing the mechanism of inhalation injury on the level of gene expression.