Expression of long non-coding RNA RP13-349O20.2 in cervical cancer tissues and experimental study of its effect on migration, invasion abilities and chemosensitivity of cervical cancer cells / 肿瘤研究与临床

Objective:To explore the expression of long non-coding RNA (lncRNA) RP13-349O20.2 in cervical cancer tissues and its impact on the migration, invasion abilities and chemotherapy sensitivity of cervical cancer cells in vitro and the possible mechanisms.Methods:The GEPIA.CANCER website (the data was updated in June 2023) was used to analyze the relationship between the expression level of RP13-349O20.2 and the overall survival of 253 cervical cancer patients. From January 2020 to August 2022, cancer tissues and paracancerous tissues (>2 cm from the tumor edge) from 40 cervical cancer patients in the Affiliated Tengzhou Central People's Hospital of Xuzhou Medical University were retrospectively collected. Human normal cervical epithelial cells H8 and human cervical cancer cell lines HCC94, C33A, Hela, HCC1106 and SiHa were used for cell experiments in vitro. Real-time fluorescence polymerase chain reaction (qRT-PCR) was used to detect the relative expression of RP13-349O20.2 in cervical cancer tissues, paracancerous tissues and each cell line. The C33A cells with the highest relative expression level of RP13-349O20.2 were transfected with small interfering RNA (siRNA) of RP13-349O20.2 and siRNA of its negative control sequence, and they were si-RP13-349O20.2 group and si-Con group, respectively. The scratch healing assay was used to detect the migration ability of C33A cells in the two groups, the Transwell assay was used to detect the invasion ability of C33A cells, and the CCK-8 method was used to detect the sensitivity of C33A cells to 5-fluorouracil. The absorbance value indicated the cell proliferation ability, the lower the absorbance value, the weaker the proliferation ability, the more sensitive to the drug. Dual-luciferase reporter gene assay was used to verify the targeting relationship between RP13-349O20.2 and miRNA-493-5p (miR-493-5p), miR-493-5p and Nectin-4. qRT-PCR was used to detect the relative expression of miR-493-5p and Nectin-4 mRNA in two groups of C33A cells, and Western blotting was used to detect the expressions of Nectin-4 protein and PI3K-AKT signaling pathway proteins in two groups of cells.Results:Analysis based on data from GEPIA.CANCER website shows that patients with low expression of RP13-349O20.2 had better overall survival than patients with high expression ( P < 0.01). The relative expression levels of RP13-349O20.2 in cervical cancer tissues and paracancerous tissues of 40 patients were 4.04±0.32 and 1.18±0.14, and the difference was statistically significant ( t = 8.29, P < 0.01). Compared with H8 cells, the expressions of RP13-349O20.2 in human cervical cancer cell lines HCC94, C33A, Hela, HCC1106 and SiHa were higher (all P < 0.01). The relative expression levels of RP13-349O20.2 in C33A cells in the si-Con group and si-RP13-349O20.2 group were 7.30±0.30 and 1.01±0.27, and the difference was statistically significant ( t = 15.62, P < 0.01). The scratch healing rates of C33A cells in the si-Con group and si-RP13-349O20.2 group were (32±9)% and (75±6)% ( t = 3.97, P < 0.01), and the numbers of invasive cells were (106±12) cells and (36±8) cells ( t = 4.79, P < 0.01). After the action of 5, 10, 20, 40 and 80 μmol/L 5-fluorouracil for 24 h, the absorbance value of C33A cells in the si-RP13-349O20.2 group was lower than that in the si-Con group. Dual-luciferase reporter gene assay confirmed that there was a targeting relationship between P13-349O20.2 and miR-493-5p ( P < 0.01), and there was a targeting relationship between miR-493-5p and Nectin-4 ( P < 0.01) . The relative expression levels of miR-493-5p in C33A cells in the si-Con group and si-RP13-349O20.2 group was 1.02±0.13 and 5.48±0.85 ( t = 5.21, P < 0.01). The relative expression levels of Nectin-4 mRNA were 5.65±0.33 and 0.99±0.34 ( t = 9.87, P < 0.01). The expression of Nectin-4 protein in C33A cells in the si-RP13-349O20.2 group was lower than that in the si-Con group ( t = 9.21, P = 0.001), and the expressions of PI3K-AKT signaling pathway proteins p-STAT3, p-PI3K, p-AKT and p-mTOR were lower than those in the si-Con group (all P < 0.01). Conclusions:The level of RP13-349O20.2 in cervical cancer tissues is high, and its high expression may indicate the poor prognosis of patients. Interfering with the expression of RP13-349O20.2 in vitro can inhibit the migration and invasion abilities of cervical cancer cells and promote the sensitivity of cervical cancer cells to 5-fluorouracil. The mechanism may be related to the miR-493-5p/Nectin-4 signaling pathway and the PI3K-AKT signaling pathway.

Effects of Hcy and lncRNA ANRIL on atherosclerotic inflammatory response / 中国医师杂志

Objective:To study the relationship between the level of serum homocysteine (Hcy) and the antisense non coding gene (ANRIL) of long chain non coding RNA (lncRNA) cell cycle dependent kinase inhibitor 2B gene, and the effect on Atherosclerosis inflammation, that is, the expression of interleukin-10 (IL-10) and Monocyte chemoattractant protein-1 (MCP-1) in Human umbilical vein endothelial cell (HUVEC).Methods:HUVEC was cultured in vitro and cells were treated with different concentration gradients (blank control group, 0.5, 1.0, 2.0, 5.0 mmol/L) of Hcy. The expression level of lncRNA ANRIL was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of MCP-1 and IL-10. LipoFilter transfection reagents were used to transfect shANRIL and shNC into different cells, respectively. In the above experiment, the optimal Hcy concentration (5.0 mmol/L) was selected for intervention for 24 hours. Western blot was used to detect the protein expression levels of MCP-1 and IL-10.Results:After 24 hours of intervention with different concentrations of Hcy in HUVEC, Hcy significantly damaged endothelial cells, and the higher the Hcy concentration, the more severe the cell damage. Compared with the blank control group, the Hcy intervention group showed an increase in lncRNA ANRIL and MCP-1, while IL-10 decreased (all P<0.05); As the concentration of Hcy intervention increases, IL-10 decreases, while lncRNA ANRIL and MCP-1 increased (all P<0.05). Compared with the blank control group, the Hcy group, the shNC+ Hcy group, and the shANRIL+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). Compared with the shANRIL+ Hcy group, the Hcy group and the shNC+ Hcy group had lower levels of IL-10 protein expression and higher levels of MCP-1 protein expression (all P<0.05). There was no statistically significant difference in the expression levels of IL-10 protein and MCP-1 protein between the shNC+ Hcy group and the Hcy group (all P>0.05). Conclusions:Hcy upregulates MCP-1 expression and downregulates IL-10 expression by promoting lncRNA ANRIL expression. Thus, it can promote cellular inflammatory reaction and participate in Atherosclerosis.

Clinical efficacy of autologous platelet rich gel in the treatment of type 2 diabetic foot patients and its effect on the expression of MALAT1 in peripheral blood mononuclear cells / 中国医师杂志

Objective:To observe the clinical efficacy of autologous platelet rich gel (APG) in the treatment of type 2 diabetic foot (DF) patients and the effect of APG on the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in peripheral blood mononuclear cells (PBMCs).Methods:A total of 62 patients with DF admitted to the Affiliated Hospital of Kangda College of Nanjing Medical University from February 2021 to May 2022 were randomly divided into a control group (30 cases) and an observation group (32 cases) using a random number table method. The control group received ultrasound debridement and dressing change treatment, while the observation group received ultrasound debridement combined with APG treatment. After 6 weeks of treatment, the effective rate, transcutaneous oxygen partial pressure (TcPO 2), and serum tumor necrosis factor- α (TNF-α), interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), hypoxia inducible factor α (HIF-1 α)and the level of MALAT1 expression in PBMCs of the two groups of patients were observed. The Pearson correlation analysis was used to investigate the relationship between the expression change of MALAT (△ MALAT1) and the total effective rate of treatment. Results:The total effective rate of the observation group was higher than that of the control group [93.75%(30/32) vs 73.33%(22/30), P<0.05]. After treatment, the systolic blood pressure (SBP), diastolic blood pressure (DBP), cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), fasting blood glucose (FPG), glycosylated hemoglobin (HbA 1c), urinary microalbumin/creatinine (UACR), uric acid (UA), white blood cells (WBC), TNF- α and IL-6 of both groups had decreased compared to before; HIF-1 α, VEGF and MALAT1 increased compared to before treatment (all P<0.05); After treatment, there was a statistically significant difference in UA, HIF-1α, VEGF, and MALAT1 between the observation group and the control group (all P<0.05). Pearson correlation analysis showed that Δ MALAT1 in DF patients was negatively correlated with TNF -α ( r=-0.61, P=0.02), IL-6 ( r=-0.52, P=0.04), WBC ( r=-0.53, P=0.03), and positively correlated with VEGF ( r=0.58, P=0.03) and HIF-1α ( r=0.54, P=0.03). The total effective rate of DF treatment was higher in the high change group of△ MALAT [88.37%(38/43) vs 73.68%(14/19), P<0.05]. There was no statistically significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:APG can significantly upregulate the expression of MALAT, improve wound tissue blood perfusion, wound angiogenesis, and inflammatory response, promote ulcer healing, and changes in MALAT expression can help determine the prognosis of DF.

Research progress on the role of non-coding RNA in chemotherapy resistance of osteosarcoma / 中华骨科杂志

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. With the emergence of chemotherapy resistance in recent years, the survival rate of osteosarcoma patients has reached a bottleneck. Therefore, exploration of its chemoresistance mechanism is one of the popular research directions currently. Non-coding RNA (ncRNA) is a class of RNA without the ability to encode proteins, which is classified into microRNA, long non-coding RNA, and circular RNA based on length and shape. With the development of high-throughput sequencing technology, there is increasing evidence that some non-coding RNAs are abnormally upregulated or downregulated in osteosarcoma cells and affect the response of osteosarcoma to four commonly used chemotherapeutic drugs (methotrexate, doxorubicin, cisplatin and ifosfamide) through mechanisms such as regulation of apoptosis, cell cycle, signaling pathways, intracellular drug concentration, and cellular autophagy. Therefore, thesenon-coding RNAs are expected to be novel targets for osteosarcoma treatment. In this paper, the current studies were searched and reviewed on the above three non-coding RNAs mediating chemoresistance in osteosarcoma, aiming to provide a reference for breaking the bottleneck of survival rate of osteosarcoma patients.

Role of NORAD in ketamine-induced neurotoxicity in mouse hippocampal neurons: association with endoplasmic reticulum stress / 中华麻醉学杂志

Objective:To evaluate the role of long non-coding RNA (lncRNA) NORAD in ketamine-induced neurotoxicity in mouse hippocampal neurons and the relationship with endoplasmic reticulum stress.Methods:Primary mouse hippocampal neurons were isolated and cultured and then divided into 5 groups ( n=36 each) using a random number table method: control group (group C), ketamine group (group K), ketamine+ pcDNA3.1-NORAD plasmid group (group K+ NORAD), ketamine+ control plasmid group (group K+ NC), and ketamine+ NORAD+ tunicamycin group (group K+ NORAD+ TM). Group C was cultured with normal medium for 24 h. Group K was cultured with 40 μmol/L ketamine for 24 h. Group K+ NORAD was transfected with pcDNA3.1-NORAD overexpressing plasmid for 48 h, followed by treatment with 40 μmol/L ketamine for 24 h. Group K+ NC was transfected with pcDNA3.1 (+ ) plasmid for 48 h, followed by treatment with 40 μmol/L ketamine for 24 h. Group K+ NORAD+ TM was transfected with pcDNA3.1-NORAD overexpressing plasmid, 24 h later endoplasmic reticulum stress activator tunicamycin 1 μg/ml was added and the neurons were cultured for 24 h, and then ketamine 40 μmol/L was added and the neurons were cultured for another 24 h. Cell viability was detected by CCK-8 assay. The amount of lactate dehydrogenase (LDH) released was analyzed. Cell apoptosis was determined by TUNEL and flow cytometry methods. The NORAD expression was detected by real-time polymerase chain reaction. The expression of endoplasmic reticulum stress-related proteins protein kinase R-like ER kinase (PERK), phosphorylated PERK (p-PERK) and C/EBP homologous protein (CHOP) was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the amount of LDH released, percentage of apoptotic neurons and apoptosis rate were increased, NORAD expression was down-regulated, CHOP expression was up-regulated, and p-PERK/PERK was increased in group K ( P<0.05). Compared with group K, the cell viability was significantly increased, the amount of LDH released, percentage of apoptotic neurons and apoptosis rate were decreased, NORAD expression was up-regulated, CHOP expression was down-regulated, and p-PERK/PERK was decreased in group K+ NORAD ( P<0.05), and no significant change was found in the parameters mentioned above in group K+ NC ( P>0.05). Compared with group K+ NORAD, the cell viability was significantly decreased, the amount of LDH released, percentage of apoptotic neurons and apoptosis rate were increased, CHOP expression was up-regulated, and p-PERK/PERK was increased ( P<0.05), and no significant change was found in the NORAD expression in group K+ NORAD+ TM ( P>0.05). Conclusions:Over-expressed NORAD can alleviate ketamine-induced neurotoxicity in mouse hippocampal neurons via inhibition of the endoplasmic reticulum stress.

Effects of long non-coding RNA HAGLR on the prognosis of breast cancer and the construction of competitive endogenous RNA interaction network / 肿瘤研究与临床

Objective:To explore the expression of long non-coding RNA (lncRNA) HAGLR in breast cancer and its effect on the prognosis of breast cancer, and to construct a competitive endogenous RNA (ceRNA) network.Methods:The Atlas of Genetics and Cytogenetics in Oncology and Haematology website was used to search for HAGLR chromosome gene mapping and transcript expression. The lnclocater website was used to predict the subcellular localization of HAGLR, and the differential expression of HAGLR in breast cancer tissues and adjacent tissues was analyzed by using lnCAR database. The patients in lnCAR database were divided into HAGLR high expression group and HAGLR low expression according to HAGLR expression. The Kaplan-Meier method was used to analyze the overall survival (OS) and metastasis-free survival, which was verified by using UCSC Xena database. lnCAR database was used to search the co-expressed genes of HAGLR. The top 200 co-expressed genes were submitted to the Metascape website for Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis, and protein interaction network (PPI) was constructed. Starbase, a bioinformatics online analysis website, was used to predict HAGLR targeting mircoRNA (miRNA) and mRNA that directly encoded proteins. ceRNA network of HAGLR was constructed with Cytoscape3.8 software.Results:HAGLR gene was localized in 2q31.1 and mainly distributed in cytoplasm. The expression level of HAGLR in breast cancer tissues was higher than that in adjacent tissues, and the difference was statistically significant ( P < 0.001). lnCAR database and UCSC Xena database analysis showed that OS in HAGLR high expression group was worse than that in HAGLR low expression group (all P < 0.01). lnCAR database, the metastasis-free survival in HAGLR high expression group was worse than that in HAGLR low expression group ( P = 0.030). Among the top 200 HAGLR co-expressed genes, 129 genes were negatively correlated with HAGLR and 71 genes were positively correlated with HAGLR. KEGG pathway analysis showed that HAGLR was related to metabolic pathways, MAPK signaling pathway, JAK-STAT signaling pathway and cancer pathway. GO annotation analysis showed that HAGLR was mainly enriched in cell cycle, centromeric complex assembly, mitotic progression, protein kinase binding, kinase activity regulation, cell response to DNA damage stimulation and other functions. hsa-miR-130b-3p, hsa-miR-1245b-5p, hsa-miR-182b-5p, hsa-miR-512-3p, hsa-miR-302b-3p, hsa-miR-185b-5p, hsa-miR-106b-5p were HAGLR targeting miRNA. Conclusions:HAGLR is highly expressed in breast cancer tissues, and it may be a biomarker for predicting the prognosis of breast cancer.

Study progression on mechanism of long non-coding RNAs regulating drug resistance in osteosarcoma / 国际肿瘤学杂志

Osteosarcoma is the most common primary solid bone malignancy. The main factor leading to recurrence and metastasis of osteosarcoma is resistance to chemotherapy drugs. Long non-coding RNAs can affect drug resistance in osteosarcoma by regulating epithelial-mesenchymal transition, cell autophagy, apoptosis, drug efflux, and cell cycle, suggesting that long non-coding RNAs may become new targets for drug resistance in osteosarcoma treatment.

Role of E2F1 and lncRNAs in the development of malignant tumors / 国际肿瘤学杂志

E2F1, a nucleoprotein gene belongs to transcription factor, is closely associated with the development of malignant tumours. Long non-coding RNAs (lncRNAs) are aberrantly expressed in a variety of tumors. In studies of molecular mechanisms associated with lncRNAs and tumours, E2F1 has been identified as a key factor that can play a critical role as an upstream regulator or downstream target of lncRNAs, and even inter-regulate to form a positive feedback loop. This paper reviews the significance of the interaction between E2F1 and lncRNA in malignant tumors in recent years, and aims to provide ideas for the study of tumor mechanisms.

lncRNA FTX silencing inhibits gastric cancer cell proliferation through the miR-22-3p/NLRP3 inflammasome pathway / 国际肿瘤学杂志

Objective:To investigate the regulatory effect of long non-coding RNA (lncRNA) FTX on gastric cancer cell proliferation through miR-22-3p/NOD-like receptor protein 3 (NLRP3) inflammasome pathway.Methods:The gastric cancer cell line NCI-N87 were divided into blank control group, si-FTX-NC group, si-FTX group, si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group. Quantitative real-time fluorescent PCR was performed to analyze the expression levels of lncRNA FTX and miR-22-3p, clone formation assay was performed to analyze the proliferation ability of NCI-N87 cells, western blotting was performed to analyze the expressions of NLRP3 inflammasome pathway proteins, and dual-luciferase reporter assay was performed to analyze the targeting relationship between lncRNA FTX and miR-22-3p.Results:The relative expressions of lncRNA FTX in the blank control group, si-FTX-NC group, si-FTX group, si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group were 1.03±0.09, 1.01±0.15, 0.42±0.08, 0.45±0.06 and 0.46±0.13 respectively, with a statistically significant difference ( F=52.19, P<0.001). The relative expressions of miR-22-3p were 1.04±0.12, 0.97±0.08, 2.26±0.15, 2.23±0.13 and 1.15±0.11 respectively, with a statistically significant difference ( F=178.53, P<0.001). Compared with the blank control group and si-FTX-NC group, the relative expressions of lncRNA FTX in the si-FTX group, si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group decreased (all P<0.001). Compared with the blank control group, si-FTX-NC group and si-FTX+miR-22-3p inhibitor group, the relative expressions of miR-22-3p in the si-FTX group and si-FTX+miR-22-3p inhibitor-NC group increased (all P<0.001). The clones of the five groups were 115.50±7.25, 112.33±8.46, 54.83±5.17, 56.17±6.32 and 85.67±9.43, with a statistically significant difference ( F=91.67, P<0.001). The levels of NLRP3 protein in the five groups were 1.84±0.17, 1.86±0.12, 0.95±0.09, 0.97±0.11 and 1.28±0.19, with a statistically significant difference ( F=60.62, P<0.001). Compared with the blank control group and si-FTX-NC group, the number of clones and the level of NLRP3 protein of the si-FTX group, si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group decreased (all P<0.05). Compared with the si-FTX+miR-22-3p inhibitor group, the number of clones and the level of NLRP3 protein in the si-FTX group and si-FTX+miR-22-3p inhibitor-NC group decreased (all P<0.05). The dual-luciferase reporter assay found that miR-22-3p was the target gene of lncRNA FTX. Conclusion:Silencing the expression of lncRNA FTX can inhibit the proliferation of gastric cancer cells, and the mechanism may be related to the regulation of lncRNA FTX on the miR-22-3p/NLRP3 inflammasome pathway.

SLC26A4-AS1 Agrava a Hipertrofia Cardíaca Induzida por AngII Aumentando a Expressão de SLC26A4 / SLC26A4-AS1 Aggravates AngII-induced Cardiac Hypertrophy by Enhancing SLC26A4 Expression

Resumo Fundamento Foi relatado que o RNA 1 antisenso 1 (SLC26A4-AS1) do membro 4 da família de transportadores de soluto 26 está altamente relacionado à hipertrofia cardíaca. Objetivo Esta pesquisa visa investigar o papel e o mecanismo específicos de SLC26A4-AS1 na hipertrofia cardíaca, fornecendo um novo marcador para o tratamento da hipertrofia cardíaca. Métodos Angiotensina II (AngII) foi infundida em cardiomiócitos ventriculares (NMVCs) de camundongos neonatos para induzir hipertrofia cardíaca. A expressão gênica foi detectada por PCR quantitativo em tempo real (RT-qPCR). Os níveis de proteína foram avaliados por western blot. Ensaios funcionais analisaram o papel de SLC26A4-AS1. O mecanismo de SLC26A4-AS1 foi avaliado por imunoprecipitação de proteína de ligação a RNA (RIP), pull-down de RNA e ensaios de luciferase repórter. O valor de p < 0,05 foi identificado como significância estatística. O teste t de Student avaliou a comparação dos dois grupos. A diferença entre os diferentes grupos foi analisada por análise de variância (ANOVA) de uma via. Resultados SLC26A4-AS1 é regulado para cima em NMVCs tratados com AngII e promove hipertrofia cardíaca induzida por AngII. SLC26A4-AS1 regula o membro 4 da família de transportadores de soluto 26 (SLC26A4) por meio do funcionamento como um RNA endógeno competitivo (ceRNA) para modular o microRNA (miR)-301a-3p e o miR-301b-3p em NMVCs. SLC26A4-AS1 promove hipertrofia cardíaca induzida por AngII via regulação para cima de SLC26A4 ou absorção de miR-301a-3p/miR-301b-3p. Conclusão SLC26A4-AS1 agrava a hipertrofia cardíaca induzida por AngII via absorção de miR-301a-3p ou miR-301b-3p para aumentar a expressão de SLC26A4.

Abstract Background It has been reported that solute carrier family 26 members 4 antisense RNA 1 (SLC26A4-AS1) is highly related to cardiac hypertrophy. Objective This research aims to investigate the role and specific mechanism of SLC26A4-AS1 in cardiac hypertrophy, providing a novel marker for cardiac hypertrophy treatment. Methods Angiotensin II (AngII) was infused into neonatal mouse ventricular cardiomyocytes (NMVCs) to induce cardiac hypertrophy. Gene expression was detected by quantitative real-time PCR (RT-qPCR). Protein levels were evaluated via western blot. Functional assays analyzed the role of SLC26A4-AS1. The mechanism of SLC26A4-AS1 was assessed by RNA-binding protein immunoprecipitation (RIP), RNA pull-down, and luciferase reporter assays. The P value <0.05 was identified as statistical significance. Student's t-test evaluated the two-group comparison. The difference between different groups was analyzed by one-way analysis of variance (ANOVA). Results SLC26A4-AS1 is upregulated in AngII-treated NMVCs and promotes AngII-induced cardiac hypertrophy. SLC26A4-AS1 regulates its nearby gene solute carrier family 26 members 4 (SLC26A4) via functioning as a competing endogenous RNA (ceRNA) to modulate the microRNA (miR)-301a-3p and miR-301b-3p in NMVCs. SLC26A4-AS1 promotes AngII-induced cardiac hypertrophy via upregulating SLC26A4 or sponging miR-301a-3p/miR-301b-3p. Conclusion SLC26A4-AS1 aggravates AngII-induced cardiac hypertrophy via sponging miR-301a-3p or miR-301b-3p to enhance SLC26A4 expression.

Construction of a prognostic model of bladder cancer using cuproptosis-associated long non-coding RNA based on The Cancer Genome Atlas database / 肿瘤研究与临床

Objective:To construct a prognostic risk model of bladder cancer using cuproptosis-associated long non-coding RNA (lncRNA) and test its predictive efficacy.Methods:RNA expression sequencing data and clinical data of corresponding samples were downloaded from The Cancer Gene Atlas (TCGA) database. The 17 key genes associated with cuproptosis was obtained from the published literature, and then lncRNA of the key genes associated with cuproptosis was screened by correlation analysis based on the lncRNA data from TCGA database. The cuproptosis lncRNA associated with the prognosis of bladder cancer patients were screened by using Cox regression and Lasso regression. A total of 403 bladder cancer patients with complete clinical information screened from TCGA database were divided into a training set (203 cases) and a test set (200 cases), and the prognostic risk prediction model was constructed based on the samples in the training set and the above key independent prognosis-related cuproptosis lncRNA. According to the median value of the risk score, patients in all the datasets, the test set and the training set of bladder cancer screened from TCGA database were divided into high-risk group and low-risk group, and R language survival package was applied to compare the differences in overall survival between the two groups in each dataset. The predictive effect of the model was verified using principal component analysis (PCA) and receiver operating characteristic (ROC) curve. Univariate and multivariate Cox regression analysis were used to analyze the factors affecting overall survival of 403 bladder cancer patients, and ROC curve was used to analyze the efficacy of each factor for predicting the prognosis of bladder cancer.Results:After screening, a total of 4 cuproptosis lncRNA with independent prognostic significance were included (AC104564.3, LINC00649, AL136084.3 and AL136295.2), and the prognostic model constructed based on these 4 lncRNA was as follows: risk score = -0.713 42×AC104564.3-0.744 94×LINC00649+0.410 93×AL136084.3-0.736 89×AL136295.2. Survival analysis showed that the overall survival of the high-risk group in all datasets, the test set and the training set was poorer than that of the low-risk group (all P < 0.05), suggesting that a high risk score predicted poor prognosis. ROC curve analysis showed that the areas under the curve of applying the risk prediction model to predict 1-, 3- and 5-year overall survival of all 403 patients in TCGA database were 0.665, 0.629 and 0.692. Multivariate Cox regression analysis showed that age (≥ 65 years old vs. < 65 years old: OR = 1.027, 95% CI 1.011-1.044, P < 0.001), stage (stage Ⅳ vs. stage Ⅲ vs. stage Ⅱ vs. stage Ⅰ vs. unknown stage: OR = 1.593, 95% CI 1.308-1.939, P < 0.001) and risk score (high vs. low: OR = 1.258, 95% CI 1.126-1.406, P < 0.001) were the independent influencing factors of patients' overall survival. ROC curve analysis showed that the areas under the curve of age, stage and risk score for predicting the patients' 5-year overall survival were 0.614, 0.685 and 0.692, suggesting that the risk prediction model had better predictive efficacy. Conclusions:A prognosis risk prediction model for bladder cancer patients is constructed based on 4 lncRNA associated with cuproptosis, and the model is internally validated to have a high predictive efficacy.

Effect of LncRNA LUCAT1 targeting miR-502-5p on the biological behavior of human retinoblastoma cells / 中华实验眼科杂志

Objective:To investigate the effects of long noncoding RNA (LncRNA) lung cancer associated transcript 1 (LUCAT1) targeting microRNA (miR)-502-5p on the proliferation, migration and invasion of human retinoblastoma (RB) cells.Methods:RB tissue samples were collected from 27 RB patients who underwent eyeball enucleation in Henan Eye Hospital from May 2019 to January 2021.Another 27 normal retinal tissue specimens were collected from 12 patients with eyeball rupture, 7 with eyeball atrophy, and 8 with eyeball penetrating injury combined with pigment film incarceration who underwent the eyeball enucleation in Henan Eye Hospital during the same period.The expressions of LncRNA LUCAT1 and miR-502-5p in RB tissues, cell lines (Y-79, WERI-Rb-1, HXo-RB44) and human retinal epithelial cells (ARPE-19) were detected by real-time quantitative PCR.Y-79 RB cell was divided into control group, small interfering RNA (si)-LncRNA LUCAT1 group, si-control (con) group, pcDNA group, pcDNA-LncRNA LUCAT1 group, miR-con group, miR-502-5p group, si-LncRNA LUCAT1+ anti-miR-con group and si-LncRNA LUCAT1+ anti-miR-502-5p group, and cells in different groups were transfected with corresponding reagents.The expressions of MMP2 and MMP9 proteins were detected by Western blot.Cell proliferation activity was assayed by cell counting kit 8.Cell proliferation capability was detected by colony formation assay.Cell migration and invasion ability were determined by Transwell assay.The targeting regulation of LncRNA LUCAT1 against miR-502-5p was confirmed by dual luciferase reporter assay and real-time quantitative PCR.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2021[32]). Written informed consent was obtained from guardians of subjects.Results:LncRNA LUCAT1 expression level in RB tissue was 2.73±0.34, which was significantly higher than 1.00±0.15 in normal retinal tissue ( t=24.190, P<0.001). The miR-502-5p expression level in RB tissues was 0.42±0.06, which was significantly lower than 1.00±0.13 in normal retinal tissue ( t=21.049, P<0.001). LncRNA LUCAT1 expression level was significantly higher and the miR-502-5p expression level was significantly lower in human RB cell lines Y-79, WERI-Rb-1 and HXO-RB44 than those in ARPE-19 cells, with statistically significant differences (all at P<0.05). The LncRNA LUCAT1 expression, the relative expressions of MMP2 and MMP9 proteins, the absorbance ( A) value, and the number of proliferated, migrating and invading Y-79 cells in si-LncRNA LUCAT1 group were significantly reduced in comparison with control group, and the differences were statistically significant (all at P<0.05). The miR-502-5p expression level was higher, and the relative expression levels of MMP2 and MMP9, A value, as well as the number of proliferated, migrating and invading Y-79 cells were lower in miR-502-5p group than in miR-con group, showing statistically significant differences ( t=20.274, 14.884, 14.181, 12.692, 17.749, 20.889, 21.913; all at P<0.001). The miR-502-5p expression level was lower and the relative expression levels of MMP2 and MMP9, A value as well as the number of proliferated, migrating and invading Y-79 cells were higher in si-LncRNA LUCAT1+ anti-miR-502-5p group than in si-LncRNA LUCAT1+ anti-miR-con group, showing statistically significant differences ( t=14.097, 15.839, 15.757, 11.860, 16.235, 16.565, 16.487; all at P<0.001). When co-transfected with LncRNA LUCAT1-wild type, the relative luciferase activity of miR-502-5p group was lower than that of miR-con group, and the difference was statistically significant ( t=16.379, P<0.001). The LncRNA LUCAT1 expression level was higher and the miR-502-5p expression level was lower in pcDNA-LncRNA LUCAT1 group than in pcDNA group, and the differences were statistically significant (both at P<0.05). The LncRNA LUCAT1 expression level was lower and the miR-502-5p expression level was higher in si-LncRNA LUCAT1 group than in si-con group, and the differences were statistically significant (both at P<0.05). Conclusions:Inhibition of LncRNA LUCAT1 can attenuate the proliferation, migration and invasion ability of human RB cells by the targeting up-regulation of miR-502-5p.

Role of linc01410 in the occurrence and development of malignant tumors / 国际肿瘤学杂志

Long non-coding RNA (lncRNA) is a class of highly conserved transcript with a length of more than 200 nucleotides, which is of great significance for the occurrence, development, diagnosis and treatment of malignant tumors. The abnormal expression of linc01410 in malignant tumors can affect the occurrence and development of malignant tumors by regulating the biological processes such as proliferation, migration and epithelial-mesenchymal transformation of malignant tumor cells, acting on related signaling pathways such as nuclear factor-κB and Notch or through exosome pathways.

SPRY4-IT1 and breast cancer / 国际肿瘤学杂志

Long non-coding RNA (lncRNA) is an RNA molecule that does not code to express proteins, and plays an important role in the occurrence and development of a variety of tumors. As an lncRNA, SPRY4-IT1 is highly expressed in breast cancer tissues, and can be used as an upstream and downstream regulator of breast cancer, promoting the progression of breast cancer, and is closely related to breast cancer stage and prognosis. In-depth study of the molecular mechanism associated with SPRY4-IT1 and breast cancer can provide new ideas for discovering biomarkers for early diagnosis of breast cancer, assessing disease prognosis and finding targeting sites.

The effect of LncRNA-MIAT mediated miR-205 expression on acute kidney injury in septic rats / 中国医师杂志

Objective:To investigate the effect of long chain non coding RNA (LncRNA)-myocardial infarction associated transcript (MIAT) mediated expression of microRNA-205 (miR-205) on acute kidney injury in septic rats.Methods:A total of 32 clean grade Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a transfection control group, and a transfection group, with 8 rats in each group. Except for the normal group, other groups of rats were used to establish sepsis acute kidney injury models by cecal ligation and puncture method. The control group was transfected with non-specific small interfering RNA (siRNA), while the transfection group was transfected with LncRNA-MIAT specific siRNA. The expression of miR-205 in renal tissue was measured using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) method; The serum levels of creatinine (Scr) and urea nitrogen (BUN) were measured using a fully automated biochemical analyzer; The levels of serum interleukin-6 (IL-6) and tumor necrosis factor -α (TNF -α) were measured using enzyme-linked immunosorbent assay (ELISA); In situ end labeling (TUNEL) method was used to determine apoptosis in rat kidney tissue cells.Results:Compared with the normal group, the model group, transfected control group, and transfected group had higher relative expression levels of miR-205 (all P<0.05); The relative expression level of miR-205 in the transfection group was lower than that in the model group and transfection control group (all P<0.05). Compared with the normal group, the model group, transfection control group, and transfection group had higher serum Scr and BUN levels (all P<0.05); The serum Scr and BUN levels in the transfection group were lower than those in the model group and transfection control group (all P<0.05). Compared with the normal group, the model group, transfection control group, and transfection group had higher levels of serum IL-6 and TNF -α (all P<0.05); The serum IL-6 and TNF-α levels in the transfection group were lower than those in the model group and the transfection control group (all P<0.05). Compared with the normal group, the model group, transfection control group, and transfection group had higher rates of renal tissue cell apoptosis (all P<0.05); The apoptosis rate of renal tissue cells in the transfection group was lower than that in the model group and transfection control group (all P<0.05). Conclusions:LncRNA-MIAT exerts a renal protective effect on acute kidney injury in septic rats by regulating the expression of miR-205. Its mechanism may be related to downregulating miR-205 expression, reducing inflammatory response, and reducing cell apoptosis in renal tissue.

Research progress of LncRNA NEAT1 in lung diseases / 中国医师杂志

Long chain non coding RNA (LncRNA) is widely involved in various biological processes such as intracellular chromatin modification, transcriptional regulation, nuclear transport, and protein function regulation, and is closely related to various key physiological functions such as immunity and metabolism in the body. NEAT1 (nuclear parapackle assembly transcript 1) is a newly discovered LncRNA, which is an important component of the nuclear substructural paraplaques. It has been proven to regulate downstream protein expression by binding to various miRNAs, thereby regulating the expression of inflammatory factors, epithelial mesenchymal transition, autophagy, apoptosis, proliferation, migration, and other biological processes, Its abnormal expression plays an important role in the pathogenesis of lung diseases such as asthma, chronic obstructive pulmonary disease, pneumonia, pulmonary fibrosis, and lung cancer, and is closely related to the prognosis of non-small cell lung cancer and the sensitivity of anti-tumor drugs. It is expected to become a new biological marker and therapeutic intervention target. This article mainly reviews the latest research progress on the role of NEAT1 in lung diseases.

Expression of long non-coding RNA LINC00665 in hepatocellular carcinoma and its effect on angiogenesis and mechanism / 中华肝胆外科杂志

Objective:To investigate the expression of long non-coding RNA 00665 (LINC00665) in hepatocellular carcinoma (HCC) and its regulatory effect on angiogenesis of hepatocellular carcinoma cells and its potential molecular mechanism.Methods:HCC tissues and corresponding paracancerous tissues of 100 patients with HCC in the First Affiliated Hospital of Nanjing Medical University from May 2016 to April 2017 were collected, and the survival prognosis was compared. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of LINC00665 in HCC tissues and cells. The effect of LINC00665 overexpressed Hep-3B cells on the angiogenesis of human umbilical vein endothelial cells (HUVEC) was examined by tube formation assay and chick chorioallantoic membrane assay. Bioinformatics database predicted the downstream microRNA (miRNA) and target genes of LINC00665, and the relationship between LINC00665, miR-126-5p and vascular endothelial growth factor A (VEGFA) was verified by RT-qPCR, Western blot and dual-luciferase reporter gene assay.Results:The expression level of LINC00665 in HCC (6.5±2.8) was significantly higher than that in paracancer tissues (4.8±3.1), the difference was statistically significant ( t=4.12, P<0.001). According to the median LINC00665 expression level of 100 patients with HCC, the cumulative survival rate of LINC00665 high expression group ( n=50) was lower than that of LINC00665 low expression group ( n=50), and the difference was statistically significant (χ 2=3.79, P=0.008). After co-culture with LINC00665 group (Hep-3B cells overexpressing LINC00665), the length of HUVEC cell tubule formation was (596.0±22.3) μm, and the number of HUVEC cell tubules was (36.3±4.5), which were both higher than NC group with the tubule formation length (127.0±13.5) μm and the number (9.3±1.5) of HUVEC cells co-cultured with Hep-3B cells of control group, and the differences were statistically significant ( t=31.15, 9.82, P<0.001, P=0.001). The chick chorioallantoic membrane assay results were similar to tube formation assay. Western blot detected that the relative expression of VEGFA in LINC00665 group was higher than that in NC group, the difference was statistically significant ( t=7.15, P<0.001). StarBase and DIANA database were used to predict and screen LINC00665 downstream miR-126-5p. StarBase database was used to predict the binding sites of LINC00665/miR-126-5p/VEGFA axis. In dual-luciferase reporter gene assay, the fluorescence intensity of LINC00665 and VEGFA vector co-transfected with miR-126-5p mimics decreased. Conclusion:LINC00665 is highly expressed in HCC and is associated with poor prognosis in patients with HCC. LINC00665 promotes angiogenesis of HCC by regulating the miR-126-5p/VEGFA axis.

Expression of long non-coding RNA NR_002578, NR_038264 and NR_046252 in peripheral blood mononuclear cells of patients with primary gout and its clinical significance / 中华风湿病学杂志

Objective:To explore the three long non-coding RNA (long non-coding ribonucleic acid, the expression of lncRNA NR_002578, NR_038264 and NR_046252) in peripheral blood mononuclear cells (PBMCs) of patients with primary gout arthritis (GA) and their clinical value.Methods:Peripheral venous blood, clinical data and laboratory data were collected from 60 gout patients (including 30 AG patients in acute stage and 30 IG patients in intermittent stage) and 50 healthy subjects (HC group). Quantitative reverse transcription PCR (RT-qPCR) was used to detect the expression levels of PBMCs of 3 lncRNAs in GA and HC groups, and the differences of 3 lncRNAs expression levels in different groups were compared and the correlation analysis was conducted with clinical indicators. Receiver operating characteristic curve (ROC) was constructed to evaluate the possible efficacy of lncRNAs in gout diagnosis. The measurement data conforming to normal distribution were tested by t test or variance analysis, and non-normal distribution were tested by Mann-Whitney U test or Kruskal-Wallis H test. The comparison among the three groups was conducted by SNK. Results:① The expression of NR_002578 in GA was significantly lower than that in HC [60.2(16.8, 100.1)×10 -3vs 149.5 (92.6, 221.8)×10 -3, Z=-5.75, P<0.001], subgroup analysis showed that the expression of NR_002578 in AG was significantly lower than that in IG and HC [34.3(8.6, 72.8)×10 -3vs 88.3(47.7, 109.6)×10 -3vs 149.5(92.6, 221.8)×10 -3, H=40.12, P<0.001], and lower in IG than that in HC ( P<0.001). The expression of NR_046252 in GA was significantly higher than that in HC [6.5(2.1, 21.5)×10 -3vs 2.1(1.2, 3.5)×10 -3, Z=-4.21, P<0.001]. The expression of NR_046252 in AG and IG were higher than that of the HC group [6.3(2.0, 12.0)×10 -3vs 7.2(2.4, 30.6)×10 -3vs 2.1(1.2, 3.5)×10 -3, H=21.33, P<0.001], but there was no significant difference between the AG and IG group ( P>0.05). ② Spearman correlation analysis showed that NR_002578 expression was negatively correlated with erythrocyte sedimentation rate (ESR) ( r=-0.29, P=0.024)and hypersensitive C-reactive protein (hs-CRP) ( r=-0.35, P=0.006) in gout patients. ③ The areas under ROC curve of NR_002578 and NR_046252 for diagnosing gout were 0.819 and 0.750, respectively. Conclusion:The abnormal expression of NR_002578 and NR_046252 in gout patients suggests that NR_002578 may be involved in the pathogenesis of gout.

Research progress on lncRNAs as members of ceRNA network in melanoma / 国际肿瘤学杂志

In recent years, there has been an increasing number of studies on long non-coding RNAs (lncRNAs) as a competing endogenous RNA (ceRNA) in melanoma. Different lncRNAs show high or low expression in melanoma, and competitively combine with miRNAs through ceRNA mechanism, affecting the expression of downstream target mRNAs, thereby playing the role of oncogenes or tumor suppressor genes. Understanding the role of lncRNA as ceRNA in melanoma can provide new ideas for the diagnosis and treatment of melanoma in the future.

Research progress of lncRNA and Traditional Chinese Medicine / 国际中医中药杂志

Long non-coding RNA (lncRNA) contains rich information and functions. The research on Traditional Chinese Medicine (TCM) targeting lncRNA mainly involves tumors, cardiovascular diseases, endocrine and metabolic related diseases, osteoporosis and other diseases, which are used to explore the mechanism of TCM and the differetiation of TCM syndromes or constitution, etc. LncRNA has important application prospects in the field of TCM. The study of lncRNA may provide new ways and technical methods for the research of modern TCM.