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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.
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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.
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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.
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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.
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Objective To investigate the correlations between expression of CASC2 and hepatocellular carcinoma(HCC) prognosis.Methods A total of 129 patients including 80 males and 49 females with HCC were includedin this study,ranging from 21 to 73 years in Xuanwu Hospital of Capital Medical University and Beijing You'an Hospital were retrospectively analyzed from September 2007 to January 2014.Expression of CASC2 was assessed using reverse transcription quantitative-polymerase chain reaction in HCC tissue and the adjacent normal tissue.The correlations between CASC2 mRNA level and clinicopathological parameters was investigated.The relationship between the expression of CASC2 and the prognosis of patients with HCC was analyzed by Kaplan-Meier method.A log-rank analysis was performed to identify group differences.Univariate and multivariate Cox analysis were used to analyze the variables affecting the patient's prognosis.Results In 129 HCC samples,the level of CASC2 expression (0.84 ± 0.05) was lower than (3.35 ± 0.11) adjacent normal tissue (P ＜ 0.05).There were significant differences between CASC2 expression and tumor size,histological differentiation,and tumor stage in 129 HCC speciments.The median expression level of CACS2 in HCC tissues,0.84-fold,was used as the cut-off value to divide the 129 patients into two groups:low-expression group (n =72) and high-expression group (n =57).Overall survival rate of HCC patients with high CACS2 expression was significantly higher than those of patients with low CACS2 expression(P ＜0.05).Multivariate analysis indicated that histological differentiation (HR =0.20,95％ CI:0.05 ～ 0.59),tumor stage (HR =1.71,95％ CI:1.02 ～ 2.99) and CACS2 expression (HR =O.51,95％ CI:O.08 ～0.92) were an independent predictor of overall survival.Conclusion Low expression of CACS2 might be associated with the occurrence and development of HCC.