OBJECTIVES: To explore the molecular mechanism by which lncRNA SNHG15 regulates proliferation, invasion and migration of lung adenocarcinoma cells. METHODS: The lncRNA microarray chip dataset GSE196584 and LncBase were used to predict the lncRNAs that interact with miR-30b-3p, and their association with patient prognosis were investigated using online databases, after which lncRNA nucleolar RNA host gene 15 (SNHG15) was selected for further analysis. The subcellular localization of lncRNA SNHG15 and its expression levels in normal human lung epithelial cells and lung adenocarcinoma cell lines were detected using fluorescence in situ hybridization and qRT-PCR. In cultured A549 cells, the changes in cell proliferation, migration, and invasion following transfection with a SNHG15 knockdown plasmid (sh-SNHG15), a miR-30b-3p inhibitor, or their co-transfection were assessed with EdU, wound healing, and Transwell assays. Bioinformatics analyses were used to predict the regulatory relationship between lncRNA SNHG15 and COX6B1, and the results were verified using Western blotting and rescue experiments in A549 cells transfected with sh-SNHG15, a COX6B1-overexpressing plasmid, or both. RESULTS: LncRNA SNHG15 was shown to target miR-30b-3p, and the former was highly expressed in lung adenocarcinoma, and associated with a poor patient prognosis. LncRNA SNHG15 was localized in the cytoplasm and expressed at higher levels in A549 and NCI-H1299 cells than in BEAS-2B cells. In A549 cells, lncRNA SNHG15 knockdown significantly inhibited cell migration, invasion and proliferation, and these changes were reversed by miR-30b-3p inhibitor. A regulatory relationship was found between lncRNA SNHG15 and COX6B1, and their expression levels were positively correlated (r=0.128, P=0.003). MiR-30b-3p knockdown obviously decreased COX6B1 expression in A549 cells, and COX6B1 overexpression rescued the cells from the inhibitory effects of lncRNA-SNHG15 knockdown. CONCLUSIONS LncRNA SNHG15 may compete with COX6B1 to bind miR-30b-3p through a ceRNA mechanism to affect proliferation, migration, and invasion of lung adenocarcinoma cells.
Humans ; MicroRNAs/metabolism* ; RNA, Long Noncoding/genetics* ; Cell Proliferation ; Cell Movement ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung ; Neoplasm Invasiveness ; A549 Cells ; Adenocarcinoma/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor

Objective To explore the current status of minor children parenting concerns among young and middle-aged breast cancer patients and investigate the influencing factors based on a random forest model so as to provide references for clinical interventions.Methods A convenience sampling method was used to select breast cancer patients undergoing treatment in our hospital between April and December 2023.A self-designed general information questionnaire,the Chinese version of parenting concerns questionnaire(PCQ),perceived social support scale(PSSS),concern about recurrence scale(CARS),and the brief illness perception questionnaire(BIPQ)were used for the study.A random forest model and the least absolute shrinkage and selection operator(LASSO)were employed to prioritise variables and filtered by significance.The selected variables were then incorporated into the multiple linear regression analysis.Results A total of 260 patients completed the study.The score of minor children parenting concerns of young and middle-aged breast cancer patients was 51.1±6.4.The multiple linear regression analysis,which included variables determined by random forest and LASSO regression(and sorted by the importance of influencing factors),showed that higher disease perception,lower perceived social support,greater concern about cancer recurrence,stage IV tumors,being divorced/widowed,and having more minor children were associated with higher parenting concerns among young and middle-aged breast cancer patients(all P<0.05),accounting for 57.0%of the total variance.Conclusion The minor children parenting concerns in young and middle-aged breast cancer patients are at a moderately high level and are influenced by a variety of factors.Healthcare professionals should develop targeted measures and interventions to reduce the parenting concerns among the patients.

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective To investigate the mechanism of action of the Astragalus-Chinese yam combination in treating cancer-related fatigue(CRF)in mice.Methods The active components and related targets of Astragalus-Chinese yam were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform.CRF-associated targets were identified using the GeneCards database.Intersecting targets were analyzed using the DAVID database for gene ontology and kyoto encyclopedia of genes and genomes enrichment analyses.A network diagram depicting"drug-active component-intersecting targets-disease"was constructed using Cytoscape software,and a protein-protein interaction network was created to identify the top five core target proteins based on degree values.Molecular docking simulations were performed using Autodock Vina software.Twenty-five mice were divided randomly into a blank group and a modeling group in a 1∶4 ratio.After successfully establishing the CRF model using Lewis lung cancer cells,mice in the modeling group were further divided into model,Chinese yam(0.2 g/kg),Astragalus(0.6 g/kg),and Astragalus-Chinese yam combination groups(0.3+0.1 g/kg)(n=5 mice per group).The treatments were administered by gavage twice daily for 14 consecutive days.Grip-strength and forced-swimming tests were conducted.The mice were then euthanized and tissues were collected.The gastrocnemius muscles were weighed and stained with hematoxylin and eosin to reveal the muscle fiber morphology.Results A total of 23 effective active components of Astragalus-Chinese yam were identified through network pharmacology analysis,with 199 intersecting drug-disease targets.These targets mainly participated in biological processes such as protein phosphorylation through cellular components(cytoplasm,membrane,nucleus)and performed molecular functions such as protein binding.A total of 155 signaling pathways,including pathway in cancer and the phosphatidylinositol 3-kinase-protein kinase B signaling pathway,were involved in CRF.The critical targets of Astragalus-Chinese yam for CRF included serine/threonine kinase,tumor necrosis factor,epidermal growth factor receptor,B-cell lymphoma 2,and caspase 3.The active components quercetin and diosgenin interacted with the highest number of targets and demonstrated binding energies＜-5.0 kJ/mol with the five core targets,indicating strong ligand-receptor binding affinity.Mice in the Chinese yam and Astragalus groups exhibited increased grip strength and prolonged swimming times compared with the model group.Gastrocnemius muscle volume and mass were increased,with well-organized muscle fibers and clear boundaries,and the effects were even more pronounced in the Astragalus-Chinese yam combination group.Conclusions Astragalus-Chinese yam treats CRF via a multi-target,multi-pathway approach,enhancing muscle strength and endurance in mice,improving gastrocnemius muscle volume and mass,and alleviating muscle atrophy,thereby mitigating the associated symptoms of CRF in mice.

Objective To investigate the mechanism of action of the Astragalus-Chinese yam combination in treating cancer-related fatigue(CRF)in mice.Methods The active components and related targets of Astragalus-Chinese yam were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform.CRF-associated targets were identified using the GeneCards database.Intersecting targets were analyzed using the DAVID database for gene ontology and kyoto encyclopedia of genes and genomes enrichment analyses.A network diagram depicting"drug-active component-intersecting targets-disease"was constructed using Cytoscape software,and a protein-protein interaction network was created to identify the top five core target proteins based on degree values.Molecular docking simulations were performed using Autodock Vina software.Twenty-five mice were divided randomly into a blank group and a modeling group in a 1∶4 ratio.After successfully establishing the CRF model using Lewis lung cancer cells,mice in the modeling group were further divided into model,Chinese yam(0.2 g/kg),Astragalus(0.6 g/kg),and Astragalus-Chinese yam combination groups(0.3+0.1 g/kg)(n=5 mice per group).The treatments were administered by gavage twice daily for 14 consecutive days.Grip-strength and forced-swimming tests were conducted.The mice were then euthanized and tissues were collected.The gastrocnemius muscles were weighed and stained with hematoxylin and eosin to reveal the muscle fiber morphology.Results A total of 23 effective active components of Astragalus-Chinese yam were identified through network pharmacology analysis,with 199 intersecting drug-disease targets.These targets mainly participated in biological processes such as protein phosphorylation through cellular components(cytoplasm,membrane,nucleus)and performed molecular functions such as protein binding.A total of 155 signaling pathways,including pathway in cancer and the phosphatidylinositol 3-kinase-protein kinase B signaling pathway,were involved in CRF.The critical targets of Astragalus-Chinese yam for CRF included serine/threonine kinase,tumor necrosis factor,epidermal growth factor receptor,B-cell lymphoma 2,and caspase 3.The active components quercetin and diosgenin interacted with the highest number of targets and demonstrated binding energies＜-5.0 kJ/mol with the five core targets,indicating strong ligand-receptor binding affinity.Mice in the Chinese yam and Astragalus groups exhibited increased grip strength and prolonged swimming times compared with the model group.Gastrocnemius muscle volume and mass were increased,with well-organized muscle fibers and clear boundaries,and the effects were even more pronounced in the Astragalus-Chinese yam combination group.Conclusions Astragalus-Chinese yam treats CRF via a multi-target,multi-pathway approach,enhancing muscle strength and endurance in mice,improving gastrocnemius muscle volume and mass,and alleviating muscle atrophy,thereby mitigating the associated symptoms of CRF in mice.

Objective To explore the current status of minor children parenting concerns among young and middle-aged breast cancer patients and investigate the influencing factors based on a random forest model so as to provide references for clinical interventions.Methods A convenience sampling method was used to select breast cancer patients undergoing treatment in our hospital between April and December 2023.A self-designed general information questionnaire,the Chinese version of parenting concerns questionnaire(PCQ),perceived social support scale(PSSS),concern about recurrence scale(CARS),and the brief illness perception questionnaire(BIPQ)were used for the study.A random forest model and the least absolute shrinkage and selection operator(LASSO)were employed to prioritise variables and filtered by significance.The selected variables were then incorporated into the multiple linear regression analysis.Results A total of 260 patients completed the study.The score of minor children parenting concerns of young and middle-aged breast cancer patients was 51.1±6.4.The multiple linear regression analysis,which included variables determined by random forest and LASSO regression(and sorted by the importance of influencing factors),showed that higher disease perception,lower perceived social support,greater concern about cancer recurrence,stage IV tumors,being divorced/widowed,and having more minor children were associated with higher parenting concerns among young and middle-aged breast cancer patients(all P<0.05),accounting for 57.0%of the total variance.Conclusion The minor children parenting concerns in young and middle-aged breast cancer patients are at a moderately high level and are influenced by a variety of factors.Healthcare professionals should develop targeted measures and interventions to reduce the parenting concerns among the patients.

Objective:To study the effects of Linc00426 on the migration,invasion and proliferation of oral squamous carcinoma cells(OSCC).Methods:TCGA database was used to query the differential expression of Linc00426.RT-qPCR was used to detect the expression level of Linc00426 in normal human oral keratinocytes(NHOK)cells and various OSCC cells lines,the knockout effi-ciency of Linc00426 at 3 sites and the expression level of miR-455-5p.Transwell test and EdU test were used to detect the changes of the migration,invasion and proliferation of CAL27 cells.Bioinformatics databases was used to predict subcellular localization and downstream miRNAs and target proteins of Linc00426.Western blot assay was used to detect the changes of USP3 expression levels in CAL27 cells.Results:The expression level of Linc00426 was increased in OSCC tissue(P＜0.05),and its expression in CAL27,HN4 and HN30 cells was higher than that in NHOK cells(P＜0.05).Knocking out Linc00426 inhibited the migration,invasion,and proliferation ability of CAL27 cells(P＜0.05).The bioinformatics database indicates that the subcellular localization of Linc00426 is in the cytoplasm;Linc00426 is negatively correlated with miR-455-5p,and miR-455-5p is negatively correlated with USP3,in which there are targeted binding sites.Knocking out miR-455-5p promoted the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knocking out USP3 inhibited the migration,invasion and proliferation of CAL27 cells(P＜0.05).Knock out Linc00426 reduced the expression level of USP3,while knocking out miR-455-5p increased the expression level of USP3(P＜0.05).Conclusion:Linc00426 plays a role as an oncogene in OSCC cells and could inhibit the migration,invasion and proliferation of the cells by targeting USP3 to regulate miR-455-5p.

Objective:To develop a homogeneous management evaluation system for clinical observership teaching under the background of national first-class discipline construction.Methods:The preliminary contents of the assessment system were determined through a literature review and expert interviews, and two rounds of questionnaire-based consultation was conducted with 20 experts using the Delphi method. With the use of Excel 2016 and SPSS 26.0, we calculated the coefficient of judgement basis (Ca), the coefficient of familiarity (Cs), the coefficient of authority (Cr), and Kendall's coefficient of concordance ( W ) as well as the mean, standard deviation, and coefficient of variation of all parameters, to identify the specific items and weights for the homogeneous management evaluation system for clinical observership teaching under the background of national first-class discipline construction. Results:In the two rounds of consultation, experts were both 100.00% active in responding to the questionnaires; the coefficients of authority of experts were 0.889 and 0.935, respectively; the coefficients of familiarity were 0.856 and 0.936, respectively; the coefficients of judgment were 0.922 and 0.934, respectively; and Kendall's coefficients of concordance were 0.476 and 0.563, respectively. Finally, 7 first-level items and 21 second-level items were included in the content framework of the homogeneous management evaluation system.Conclusions:The construction process of the homogeneous management evaluation system of clinical observership teaching is complete and reliable, which can provide a reference for the homogeneous management of clinical observership teaching, but further verification and improvement are needed.

Objective By investigating the effects of miR-379-5p on the proliferation,invasion and metastasis of mouse breast cancer 4T1 cells,we aimed to provide new therapeutic targets for the clinical inhibition of breast cancer proliferation,invasion,and metastasis.Methods After plasmid transfection,4T1 cells were utilized to detect the expression of miR-379-5p using fluorescence quantitative PCR.While 5-ethynyl-2'doxyuridine(EdU)cell proliferation and Transwell assays were employed to detect changes in the proliferation and invasion ability of 4T1 cells in each group.The migration ability of 4T1 cells after overexpression and knockdown of miR-379-5p was examined by scratch healing assay.A transplanted tumor model of breast cancer was established in BABL/c mice,and the effects of overexpressing miR-379-5p on tumor growth and the number and size of lung metastases were observed.Results EdU result showed that knocking down miR-379-5p enhanced the proliferation ability of the cells compared with the control group cells,and miR-379-5p overexpression reduced the capacity of breast cancer cells to proliferate(P＜0.05).Transwell and wound healing assays showed that miR-379-5p knockdown enhanced,while miR-379-5p overexpression significantly inhibited,the invasion and migratory ability of breast cancer cells(P＜0.01).An in vivo tumorigenesis experiment with BABL/c mice showed that miR-379-5p overexpression significantly slowed the tumor growth rate(P＜0.05)and inhibited lung metastasis(P＜0.01).Conclusions MiR-379-5p plays a role in tumor gene suppression in breast cancer and inhibits the proliferation,invasion,and migration of mouse breast cancer 4T1 cells.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most common type of non-small cell lung cancer, and any change of miRNAs expression will affect the degree of target regulation, thus affecting intracellular homeostasis. This study verified that miR-186-5p could inhibit the proliferation, migration and invasion of LUAD cells by regulating PRKAA2. METHODS: Previous investigations found that the expression of miR-186-5p was markedly suppressed in LUAD. Bioinformatics method is used to predict the target protein related to ferroptosis downstream and inquire about its expression level in LUAD and its influence on the survival of patients. Double luciferase verified the binding site of PRKAA2 and miR-186-5p. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression of PRKAA2. The effects of miR-186-5p of LUAD cells as well as the mechanism by which miR-186-5p inhibits Fer-1's sensitivity to ferroptosis were confirmed by EdU, Transwell, and scratch assays. The effect of miR-186-5p on the amount of reactive oxygen species (ROS) in LUAD cells was discovered using ROS experiment. Malondialdehyde (MDA) and glutathione (GSH) experiments were used to detect the effects of miR-186-5p and PRKAA2 on ferroptosis index of LUAD cells. The concentration of lipid ROS (L-ROS) in LUAD cells were measured using the L-ROS tests to determine the effects of miR-186-5p and PRKAA2. RESULTS: The expression of PRKAA2 is up-regulated, and a high level of PRKAA2 expression was associated with a poor prognosis for patients with LUAD. Overexpression of miR-186-5p decreased the gene and protein expression of PRKAA2. By promoting ferroptosis, miR-186-5p overexpression prevented lung cancer cells from proliferating, invading, and migrating. ROS could be produced in higher amounts in LUAD cells due to miR-186-5p. Overexpression of miR-186-5p and knockdown PRKAA2 up-regulated MDA content and reduced GSH content in LUAD cells, respectively. miR-186-5p could increase the content of L-ROS and promote the ferroptosis sensitivity of LUAD cells by targeting PRKAA2. CONCLUSIONS miR-186-5p promotes ferroptosis of LUAD cells through targeted regulation of PRKAA2, thus inhibiting the proliferation, invasion and migration of LUAD.
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Humans ; Lung Neoplasms/genetics* ; Carcinoma, Non-Small-Cell Lung ; Ferroptosis/genetics* ; Reactive Oxygen Species ; Adenocarcinoma of Lung/genetics* ; MicroRNAs/genetics* ; 3,4-Methylenedioxyamphetamine ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor ; AMP-Activated Protein Kinases