ObjectiveTo observe the effect of lncRNA-metastasis-associated lung adenocarcinoma transcript 1(MALAT1)on colorectal cancer cells-induced angiogenesis, and explore the potential underlying mechanism. MethodsMALAT1 was overexpressed in colorectal cancer cells SW48 by plasmids transfection, then SW48 cells were cultured at normoxia or hypoxia conditions. The culture media was collected, and the concentration of vascular endothelial growth factor (VEGF) in the media was measured by the enzyme-linked immuno sorbent assay (ELISA), and the human umbilical vein endothelial cells (HUVEC) were incubated with the media collected above. Meanwhile, the expression of hypoxia-inducible factor-1α(HIF-1α) in SW48 cells was detected by western blot. ResultsOverexpression of MALAT1 increased the VEGF level in the culture media, normoxia:the MALAT1 group (514±32) mg/L vs. the control group (110±14) mg/L, P < 0.05; hypoxia:the MALAT1 group (928±18) mg/L vs. the control group (230±21) mg/L, P < 0.05. Meanwhile, the tube formation activity of HUVEC was enhanced, and the expression of HIF-1αwas elevated in the MALAT1 group by western blot. ConclusionOverexpression of MALAT1 could promote colorectal cancer cells-mediated angiogenesis, it may be developed as a new drug target for colorectal cancer treatment.
ObjectiveTo analyze the expression and clinical significance of cyclin-dependent kinase 1 (CDK1) in lung adenocarcinoma by bioinformatics.MethodsBased on the gene expression data of lung adenocarcinoma patients in The Cancer Genome Atlas (TCGA), the differential expression of CDK1 in lung adenocarcinoma tissues and normal lung tissues was analyzed. The expression of CDK1 gene in lung adenocarcinoma was analyzed by UALCAN at different angles. Survival analysis of different levels of CDK1 gene expression in lung adenocarcinoma was performed using Kaplan-Meier Plotter. Correlation Cox analysis of CDK1 expression and overall survival was based on clinical data of lung adenocarcinoma in TCGA. Gene set enrichment analysis was performed on gene sequences related to CDK1 expression in clinical cases. The protein interaction network of CDK1 from Homo sapiens was obtained by STRING. CDK1-related gene proteins were obtained and analyzed by the web server Gene Expression Profiling Interactive Analysis (GEPIA).ResultsBased on the analysis of TCGA gene expression data, CDK1 expression in lung adenocarcinoma was higher than that in normal lung tissues. UALCAN analysis showed that high CDK1 expression may be associated with smoking. Survival analysis indicated that when CDK1 gene was highly expressed, patients with lung adenocarcinoma had a poor prognosis. Univariate and multivariate Cox regression analysis of CDK1 expression and overall survival showed that high CDK1 expression was an independent risk factor for survival of patients with lung adenocarcinoma. Gene set enrichment analysis revealed that high CDK1 expression was closely related to DNA replication, cell cycle, cancer pathway and p53 signaling pathway.ConclusionCDK1 may be a potential molecular marker for prognosis of lung adenocarcinoma. In addition, CDK1 regulation may play an important role in DNA replication, cell cycle, cancer pathway and p53 signaling pathway in lung adenocarcinoma.
Objective To identify the immune cell-related biomarkers in lung adenocarcinoma using weighted gene co-expression network. Methods In this study, based on TCGA database, gene co-expression network was constructed in TCGA-LUAD by WGCNA package, and different gene modules were formed by clustering. At the same time, ESTIMATE analysis was performed on lung adenocarcinoma tumor samples in the TCGA-LUAD dataset. Gene enrichment pathways in the most significant related modules were evaluated by GO and KEGG assays. Candidate hub genes in the selected key modules were used to construct protein-protein interaction (PPI) network for intersection to obtain hub genes. The prognostic properties of these hub genes and patient immune cell infiltration were verified by Kaplan-Meier curve and TIMER algorithm. Multivariate Cox regression analysis was performed on the acquired hub gene and a prognostic risk model was constructed. Results In the co-expression network, we observed that the brown module was closely related to ImmuneScore, StromalScore and ESTIMATE Score. Five immune-related hub genes CD53, PLEK, SPI1, IL10RA and C3AR1 were obtained. The enrichment analysis of brown module found that module genes were mainly enriched in GO items such as innate immune response regulation and KEGG pathways such as NF-kappa B signaling pathway. In addition, the results of this study also found that the expression levels of 5 hub genes were significantly positively correlated with the infiltration abundance of immune cells. IPS and TIDE validated the immune relevance of the model. At the same time, we found that the RiskScore we established has great potential in predicting immunotherapy. Conclusion In summary, the five key genes related to immune cells obtained may provide new and effective potential targets for the immunotherapy of lung adenocarcinoma, which is also beneficial to provide personalized diagnosis and treatment strategies for patients with lung adenocarcinoma in the later stage.
ObjectiveTo explore the predictive value of CT signs of mixed ground-glass nodules in the pathological subtype and differentiation of lung adenocarcinoma. MethodsThe clinical data of 66 patients with mixed ground-glass nodules pathologically diagnosed as invasive adenocarcinoma (IAC) in the Second Department of Thoracic Surgery, the First Affiliated Hospital of Xiamen University from May to December 2021 were retrospectively analyzed, including 20 males and 46 females, aged 35-75 years. The CT findings were analyzed before operation, and the lesion profile was cut after operation to distinguish the ground-glass and solid components, and the pathological results of different positions were obtained. According to the postoperative pathological results, the patients were divided into a low-risk group (containing adherent type and no components of micropapillary subtype and solid subtype, n=16), a medium-risk group (containing niple or acinar type and no components of micropapillary subtype and solid subtype, n=38), and a high-risk group (containing micropapillary or solid subtype, n=12). The relationships between CT features and the pathological subtype and degree of differentiation were analyzed and compared. ResultsIn 66 patients with IAC, the infiltration degree of solid components was greater than that of ground-glass components. When the solid component ratio (CTR) was≥25% (sensitivity 90.2%, specificity 64.0%, P=0.005), and the average CT value was＞−283.95 HU (sensitivity 82.9%, specificity 64.0%, P=0.000), the histological grade was more inclined to medium and low differentiation. The CTR, Ki-67, average CT value and histological grade of IAC in the medium- and high-risk groups were higher than those of nodules in the low-risk group. ConclusionThe infiltration degree of solid components is higher than that of ground-glass components in IAC mixed ground-glass nodules. The pathological subtype, Ki-67 expression and histological grade of lung adenocarcinoma can be predicted according to its CT characteristics, which has important clinical significance for determining the timing of surgery.
ObjectiveTo investigate the association between the baseline 18F-FDG PET/CT SUVmax and histological subtypes of ≤2 cm early peripheral lung adenocarcinoma (cN0).MethodsWe retrospectively reviewed the clinical data of consecutive patients who received baseline 18F-FDG PET/CT and underwent anatomic lung resection for ≤2 cm early peripheral lung adenocarcinoma from 2011 to 2014 in our institute.ResultsA total of 195 patients were enrolled in this study, including 86 males and 109 females, with an average age of 59.96±9.19 years. Twenty-two patients were pathologically confirmed with lymph node metastasis. One hundred and fifty-seven patients were in the subtype group 1, which included lepidic, acinar, and papillary predominant tumors. Thirty-eight patients were in the subtype group 2, which included solid and micropapillary predominant tumors. The 5-year survival rate was 79.0% and 58.0% in the subtype group 1 and subtype group 2, respectively (P=0.006). The median SUVmax was 2.00 (0.30-13.10) and 4.15 (1.20-17.90) in the subtype group 1 and subtype group 2, respectively (P=0.000). Logistic regression suggested that baseline SUVmax≥2.5 was an independent risk factor for the subtype group 2 (OR=6.635, 95%CI 2.510-17.545, P=0.000). The receiver operating characteristic curve suggested that the continuous SUVmax had an moderate predictive value for subtypes (area under the curve was 0.792, 95%CI 0.717-0.866).ConclusionBaseline 18F-FDG PET/CT SUVmax has certain predictive value for histological subtypes of ≤2 cm early peripheral lung adenocarcinoma.
Lung cancer is one of the malignant tumors with the greatest threat to human health, and studies have shown that some genes play an important regulatory role in the occurrence and development of lung cancer. In this paper, a LightGBM ensemble learning method is proposed to construct a prognostic model based on immune relate gene (IRG) profile data and clinical data to predict the prognostic survival rate of lung adenocarcinoma patients. First, this method used the Limma package for differential gene expression, used CoxPH regression analysis to screen the IRG to prognosis, and then used XGBoost algorithm to score the importance of the IRG features. Finally, the LASSO regression analysis was used to select IRG that could be used to construct a prognostic model, and a total of 17 IRG features were obtained that could be used to construct model. LightGBM was trained according to the IRG screened. The K-means algorithm was used to divide the patients into three groups, and the area under curve (AUC) of receiver operating characteristic (ROC) of the model output showed that the accuracy of the model in predicting the survival rates of the three groups of patients was 96%, 98% and 96%, respectively. The experimental results show that the model proposed in this paper can divide patients with lung adenocarcinoma into three groups [5-year survival rate higher than 65% (group 1), lower than 65% but higher than 30% (group 2) and lower than 30% (group 3)] and can accurately predict the 5-year survival rate of lung adenocarcinoma patients.
ObjectiveTo investigate the regulatory effect of long chain non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) adsorbing microRNA-124 (miR-124) on osteogenic differentiation of mesenchymal stem cells (MSCs).MethodsC3H10T1/2 cells derived from mouse embryos were cultured in vitro, then randomly divided into control group (group A), lncRNA MALAT1 no-load plasmid group (group B), lncRNA MALAT1 overexpression plasmid group (group C), lncRNA MALAT1 small interfering RNA (siRNA) group (group D), and lncRNA MALAT1 siRNA negative control group (group E). The cells were transfected into plasmids and siRNA, then induced to differentiate into osteoblasts. Alkaline phosphatase (ALP) and alizarin red staining were used to detect the osteogenic differentiation of cells in each group, real-time fluorescence quantitative (qRT-PCR) analysis was used to detect the expressions of lncRNA MALAT, miR-124, and osteogenesis-related genes such as Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) in each group. Double luciferase reporter gene was used to detect the targeting regulation of lncRNA MALAT1 to miR-124.ResultsThe relative contents of ALP positive cells, mineralized nodule, and the relative mRNA expressions of lncRNA MALAT1, Runx2, OPN, and OCN in group C were significantly higher than those in other groups (P<0.05), while in group D significantly lower than in other groups (P<0.05); the relative expression of miR-124 in group C was significantly lower than that in other groups(P<0.05), while in group D significantly higher than in other groups (P<0.05). There was no significant difference in these indexes between groups A, B, and E (P>0.05). The results of double luciferase reporter gene assay showed that lncRNA MALAT1 targeting down-regulated the expression of miR-124.ConclusionLncRNA MALAT1 can targeting down-regulate the expression of miR-124 and promote the osteogenic differentiation of MSCs.
Objective To investigate the effect of microRNA-27a (miR-27a) on the apoptosis of human lung adenocarcinoma cells A549 induced by lipopolysaccharide (LPS) by regulating the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) pathway, and its mechanism is discussed preliminarily. Methods The complementary binding sites of miR-27a and phosphatidylinositol-3 kinase catalytic subunit delta (PIK3CD) were analyzed by Starbase and verified by double luciferase. The A549 cells were divided into normal group, LPS group, LPS+miR-27a mimic negative control group, LPS+miR-27a mimic group, LPS+miR-27a mimic+PI3K activator group. In the LPS+miR-27a mimic negative control group, LPS+miR-27a mimic group and LPS+miR-27a mimic+PI3K activator group, the cells were transfected with miR-27a mimic negative control, miR-27a mimic and miR-27a mimic, respectively, and were cultured for 6 h. After that, the cells were cultured in complete medium for 24 h, and then, except for the normal group, the cells in the other groups were stimulated with 10 mg/L LPS for 24 h, and the PI3K activator 740 Y-P was added to the LPS+miR-27a mimic+PI3K activator group, and cells in normal group were cultured in complete medium for the same time. Real-time quantitative polymerase chain reaction was used to detect the expression level of miR-27a in cells; cell counting kit 8 was used to detect cell proliferation; Hoechst33342 staining and flow cytometry was used to detect apoptosis; autophagy of A549 cells was observed by transmission electron microscope; Western blot was used to detect the expression of PIK3CD, phosphorylated-AKT (p-AKT), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3 and microtubule-associated protein 1 light chain 3 II (LC3II) protein. Results There was a binding site between miR-27a and PIK3CD, which was verified by double luciferase. Compared with those in normal group, the expression level of miR-27a, proliferation rate and protein expression level of Bcl-2 in LPS group and LPS+miR-27a mimic negative control group were lower (P<0.05), the apoptosis rate, protein expression levels of PIK3CD, p-AKT, Bax, cleaved caspase-3, LC3Ⅱ were higher (P<0.05); compared with those in LPS group and LPS+miR-27a mimic negative control group, the expression level of miR-27a, proliferation rate and protein expression level of Bcl-2 in LPS+miR-27a mimic group were higher (P<0.05), the apoptosis rate, protein expression levels of PIK3CD, p-AKT, Bax, cleaved caspase-3, LC3Ⅱ were lower (P<0.05); compared with those in LPS+miR-27a mimic group, the expression level of miR-27a and proliferation rate in LPS+miR-27a mimic+PI3K activator group were lower (P<0.05), the apoptosis rate, protein expression levels of PIK3CD, p-AKT, cleaved caspase-3, LC3Ⅱ were higher (P<0.05). The number of cells in the normal group was more, the cells were closely arranged, the nucleus size was uniform, and the organelle structure was normal; in LPS group and LPS+miR-27a mimic negative control group, cells became round, nuclei pyknosis, formed clumps, and showed multiple round autophagic vesicles of different sizes; the number of nuclear pyknotic cells in LPS+miR-27a mimic group decreased, and the number of nuclear pyknotic cells in LPS+miR-27a mimic+PI3K activator group increased compared with LPS+miR-27a mimic group, a small number of circular autophagic vesicles were observed, but the number was different. Conclusion Overexpression of miR-27a can inhibit PI3K/Akt pathway and reduce LPS induced apoptosis of human lung adenocarcinoma cells A549, which may be related to the reduction of autophagy.