Overcoming flumatinib resistance in chronic myeloid leukaemia: Insights into cellular mechanisms and ivermectin's therapeutic potential.

Chronic myeloid leukaemia (CML) is a haematological malignancy characterized by the constitutive tyrosine kinase activity of the BCR-ABL1 fusion protein. Flumatinib, a second-generation tyrosine kinase inhibitor, has exhibited superior clinical efficacy compared to its precursor, imatinib. However, with increased clinical use, resistance to flumatinib has emerged as a significant challenge. To investigate the mechanisms of flumatinib resistance in CML, we induced the human CML cell line K562 using a flumatinib concentration gradient method in vitro, successfully establishing a flumatinib-resistant K562/FLM cell line. This cell line exhibited cross-resistance to imatinib and doxorubicin, but remained sensitive to the antiparasitic agent ivermectin, which possesses antitumoural effects. Through cellular experimentation, we explored the resistance mechanisms, which indicated that K562/FLM cells evade flumatinib cytotoxicity by enhancing autophagy, increasing the expression of membrane transport proteins, particularly P-glycoprotein, ABCC1 and ABCC4, as well as enhancing phosphorylation of p-EGFR, p-ERK and p-STAT3 proteins. Moreover, it was found that ivermectin effectively suppressed the expression of autophagy and transport proteins in K562/FLM cells, reduced the activity of the aforementioned phosphoproteins, and promoted apoptotic cell death. Collectively, the increased autophagy, higher expression of drug-efflux proteins and hyperactivation of the EGFR/ERK/STAT3 signalling pathway were identified as pivotal elements promoting resistance to flumatinib. The significant effects of ivermectin might offer a novel therapeutic strategy to overcome flumatinib resistance and optimize the treatment outcomes of CML.

LncRNA XIST Contributes to Cisplatin Resistance of Lung Cancer Cells by Promoting Cellular Glycolysis through Sponging miR-101-3p.

BACKGROUND: Non-small-cell lung carcinoma is one of the most frequently diagnosed cancers. Cisplatin (CDDP) is a currently applied standard anticancer agent for advanced lung cancers. Although effectively clinical response was achieved initially, a large fraction of lung cancer patients developed cisplatin resistance. Therefore, understanding the molecular mechanisms of chemoresistance is crucial for anti-lung cancer therapy. Long non-coding RNA (lncRNA)-X-inactive-specific transcript (XIST) has been reported to be positively associated with multiple cancers. Currently, the precise role and mechanism of XIST in cisplatin resistance of lung cancer have not been elucidated. METHODS: The expression levels of miR-101-3p and lncRNA XIST were detected by qRT-PCR. Cisplatin-resistant lung cancer cell line was established by selecting the survival cells under gradually increased cisplatin treatments. The cell proliferation was detected by MTT assay, and the cellular glucose metabolism rate was evaluated by Seahorse metabolic flux analysis and glucose uptake and lactate product assays. Glycolysis-related protein expression levels were detected by Western blot. Dual luciferase reporter was constructed to determine the lncRNA-miRNA interaction. RESULTS: Here, we report XIST is significantly upregulated in lung cancer tissues compared with normal lung tissues. In addition, cisplatin-resistant lung cancer cells displayed remarkably elevated XIST expression. We demonstrated that miR-101-3p functioned as a tumor suppressor in lung cancer and sensitized lung cancer cells to cisplatin. Bioinformatics analysis predicted miR-101-3p could be a potential target of XIST through direct binding with it as a competing endogenous RNA, which was further validated from lung tumor tissues and cell lines by luciferase assay. Intriguingly, XIST significantly promoted cellular glycolysis rate of lung cancer cells. The extracellular acidification rate, glucose uptake, and lactate product were elevated by XIST overexpression. On the contrary, miR-101-3p effectively suppressed glycolysis rate. Finally, we demonstrated silencing XIST significantly recovered miR-101-3p expression and downregulated expression of glycolysis key enzymes, a phenotype could be further overridden by miR-101-3p inhibition. CONCLUSIONS: This study reveals a new molecular mechanism for the lncRNA-XIST-promoted cisplatin resistance via sponging miR-101-3p, leading to de-repression of cellular glycolysis. Moreover, these findings warrant further in vivo investigations to study XIST as a potential target to overcome cisplatin resistance.

LncRNA FOXD3-AS1 promoted chemo-resistance of NSCLC cells via directly acting on miR-127-3p/MDM2 axis.

BACKGROUND: This study aims to investigate the mechanism underlying the high level of long non-coding RNA FOXD3-AS1 in cisplatin-resistant NSCLC cells. METHODS: Cisplatin-resistant cells were generated from A549 cells. CCK-8 were used to evaluate cell proliferation. The FOXD3-AS1, miR-127-3p, MDM2 and MRP1 mRNA expression levels were confirmed by qRT-PCR. Protein levels of MDM2 and MRP1 were determined by western blot assay. Luciferase reporter and RNA pull-down assays were evaluated the relationship between miR-127-3p and FOXD3-AS1/MDM2. In vivo tumor growth was evaluated in a xenograft nude mice model. RESULTS: FOXD3-AS1 was up-regulated in cisplatin-resistant NSCLC cells (A549/DDP and H1299/DDP cells) in comparison with their parental cell lines. Overexpression of FOXD3-AS1 promoted cisplatin-resistance in A549 and H1299 cells; while FOXD3-AS1 knockdown sensitized A549/DDP and H1299/DDP cells to cisplatin treatment. FOXD3-AS1 regulated miR-127-3p expression by acting as a competing endogenous RNA, and miR-127-3p repressed MDM2 expression via targeting the 3'UTR. MiR-127-3p overexpression and MDM2 knockdown both increased the chemo-sensitivity in A549/DDP cells; while miR-127-3p knockdown and MDM2 overexpression both promoted chemoresistance in A549 cells. Further rescue experiments revealed that miR-127-3p knockdown or MDM2 overexpression counteracted the suppressive effects of FOXD3-AS1 knockdown on chemo-resistance and MRP1 expression in A549/DDP cells. In vivo studies showed that FOXD3-AS1 knockdown potentiated the antitumor effects of cisplatin treatment. Inspection of clinical samples showed the upregulation of FOXD3-AS1 and MDM2, and down-regulation of miR-127-3p in NSCLC tissues compared to normal adjacent tissues. CONCLUSION: In conclusion, our results suggest that LncRNA FOXD3-AS1 promotes chemo-resistance of NSCLC cells via directly acting on miR-127-3p/MDM2 axis. Our findings may provide novel perspectives for the treatment of NSCLC in patients resistant to chemotherapy.

Transcriptome analysis reveals the mechanism by which spraying diethyl aminoethyl hexanoate after anthesis regulates wheat grain filling.

BACKGROUND: Diethyl aminoethyl hexanoate (DA-6), a plant growth regulator, has many beneficial effects on agricultural production. DA-6 has been applied to many plant species, but the molecular mechanism by which spraying DA-6 after anthesis regulates wheat grain filling is still unknown. RESULTS: In this study, we used four DA-6 concentrations: C0 (0 g/L), C2 (2 g/L), C4 (4 g/L), and C6 (6 g/L). The results showed that C4 and C6 led to a significantly higher 1000-grain weight and seed protein content than C0 during two wheat growing seasons. We then subjected samples at 24 days after anthesis (at which point the grain weight increased rapidly) to transcriptome analysis. Flag leaf (L), seed (S), and stem (T) samples under C6 and C0 were used for RNA-seq. The seed samples under C6 compared with C0 (S6vsS0) presented the most differentially expressed genes (DEGs; 2164). Plant hormone signal transduction (p = 1.97 × 10- 4), protein processing in the endoplasmic reticulum (ER; p = 9.04 × 10- 11) and starch and sucrose metabolism (p = 1.90 × 10- 10) pathways were the most markedly enriched pathways in the flag leaves, stems, and seeds, respectively. DEGs involved in sucrose synthesis in the flag leaves, protein processing in ER in the stems, and starch synthesis and protein processing in ER in the seeds were significantly upregulated under C6 compared with C0. CONCLUSIONS: Overall, we propose a model for spraying DA-6 after anthesis to regulate metabolic pathways in wheat, which provides new insights into wheat in response to DA-6.

Diagnosis and treatment of otomycosis in southern China.

To analyse and discuss the clinical features and pathogenic characteristics, diagnosis and treatment of patients with otomycosis in southern China. Two hundred fifty-six patients from southern China diagnosed with otomycosis were randomly separated into two groups: the drug filling group and drug smearing group. Patients in the drug filling group were first examined and then had the pathogenic secretions in their external auditory canals cleared by otoendoscopy. Then, the local antifungal cream triamcinolone acetonide clotrimazole was injected into the external auditory canal. The same treatment was undertaken 1 week later and repeated once or twice more. Patients in the drug smearing group were also treated by otoendoscopy. Then, they were told to smear their external auditory canals once per day with the antifungal cream. All cases were followed for more than 6 months after the 3- to 4-week treatment. The main symptoms and otoendoscopic examination were used to evaluate the prognosis. Aspergillus was the commonest fungus. The cure rate was 93% in the drug filling group and 81% in the drug smearing group. Otomycosis is very common in southern China, but it lacks characteristic features in its early stages. Once diagnosed, the local lesions in the external auditory canal should be cleared thoroughly using otoendoscopy, and then, the local antifungal cream is injected into external auditory canal. The cure rate can be significantly improved with the foregoing treatment.

Evaluation of MTANNs for eliminating false-positive with different computer aided pulmonary nodules detection software.

MTANN (Massive Training Artificial Neural Network) is a promising tool, which applied to eliminate false-positive for thoracic CT in recent years. In order to evaluate whether this method is feasible to eliminate false-positive of different CAD schemes, especially, when it is applied to commercial CAD software, this paper evaluate the performance of the method for eliminating false-positives produced by three different versions of commercial CAD software for lung nodules detection in chest radiographs. Experimental results demonstrate that the approach is useful in reducing FPs for different computer aided lung nodules detection software in chest radiographs.

Construction of a prognostic model based on genome-wide methylation analysis of miRNAs for hepatocellular carcinoma.

Aim: Using the methylation level of miRNA genes to develop a prognostic model for patients with hepatocellular carcinoma (HCC). Materials & methods: least absolute shrinkage and selection operator and multivariate Cox regression analyses were performed to develop a prognostic model. One miRNA in the model was selected for verification. Results: A prognostic model was developed using eight miRNAs. The areas under the curve for predicting overall survival at 1, 3 and 5 years were 0.75, 0.81 and 0.81. miR-223 was found to be hypomethylated in 160 HCC tissues, and its methylation level was associated with Barcelona Clinic Liver Cancer stages and the prognosis of patients with HCC. Conclusion: The prognostic model based on miRNA methylation levels has the capability to partially forecast the prognosis of patients with HCC.

Refractory Epistaxis: A Map is Enough!

Combined the big data from Chinese researches and our clinical experiences, we drew a concise "distributed map" of intractable epistaxis showing the concealed bleeding regions and offending vessels clearly (Figure 1). The bleeding site was located accurately according to the "distributed map," and bleeding was stopped via bipolar radiofrequency ablation under nasal endoscope without nasal packing, followed by five classic cases (Figure 2). It is our recommended precise mode of diagnosis and treatment of refractory epistaxis.

Weakly-Interactive-Mixed Learning: Less Labelling Cost for Better Medical Image Segmentation.

Common medical image segmentation tasks require large training datasets with pixel-level annotations which are very expensive and time-consuming to prepare. To overcome such limitation and achieve the desired segmentation accuracy, a novel Weakly-Interactive-Mixed Learning (WIML) framework is proposed by efficiently using weak labels. On one hand, utilize weak labels to reduce annotation time for high-quality strong labels by designing a Weakly-Interactive Annotation (WIA) part of the WIML which prudently introduces interactive learning into the weakly-supervised segmentation strategy. On the other hand, utilize weak labels and very few strong labels to achieve desired segmentation accuracy by designing a Mixed-Supervised Learning (MSL) part of the WIML which can boost the segmentation accuracy by providing strong prior knowledge during training. Besides, a multi-task Full-Parameter-Sharing Network (FPSNet) is proposed to better implement this framework. Specifically, to further reduce annotation time, attention modules (scSE) are integrated into FPSNet to improve the class activation map (CAM) performance for the first time. To further improve segmentation accuracy, a Full-Parameter-Sharing (FPS) strategy is designed in FPSNet to alleviate the overfitting of the segmentation task supervised by very few strong labels. The proposed method is validated on the BraTS 2019 and LiTS 2017 datasets, and experiments demonstrate that the proposed method WIML-FPSNet outperforms several state-of-the-art segmentation methods with minimal annotation efforts.

Expression and Prognostic Role of CXCL1 Gene in Colorectal Adenocarcinoma.

In this manuscript, we have extensively examined expression and prognosis of CXCL1 gene in colorectal adenocarcinoma (COAD) using different cases of colorectal adenocarcinoma and tissues. To verify this, protein and mRNA expressions of cxcl1 were identified through RT-PCR and immunohistochemistry in 30 cases of colorectal adenocarcinoma and adjacent tissues, which were surgically resected from January to July 2021 in our hospital, and relationship between CXCL1 mRNA and clinicopathological features and protein expression was analyzed. CXCL 1 mRNA in COAD carcinoma's expression was considerably higher than in the adjacent normal intestine. At the same time, CXCL 1 diagnostic receiver operating characteristic (ROC) curve had preferably higher value of the diagnostic for area under curve (AUC) = 0.912, 95%, COAD (P < 0.001, CI = 0.825-0.969). We have observed that CXCL1 gene was closely linked with preoperative CEA level (P=0.007) and gross tumor typing (P=0.039). Finally, we have concluded that that CXCL1 can be a possible biomarker for stress prognosis and diagnosis.

Hypomethylation status of miR-657 promoter region as biomarker for diagnosis of hepatocellular carcinoma: a retrospective study.

Background: The purpose of our study was to evaluate whether the methylation status of the miR-657 promoter region could be used as a biomarker for diagnosis of hepatocellular carcinoma (HCC), so as to find alternative biomarkers of early HCC detection. Methods: Cancerous and paired adjacent noncancerous tissues were collected from 160 patients who had been diagnosed with HCC by histopathology and received surgery. The methylation status of the miR-657 promoter region was measured using a MassARRAY Analyzer 4. Receiver operator characteristic (ROC) curve analysis was used to assess the effectiveness of miR-657 promoter region methylation status as a biomarker for diagnosis of HCC. Results: The mean methylation level of the miR-657 promoter region was significantly lower in cancerous tissues than in normal tissues of HCC patients (48.91%:67.04%, P<0.0001). ROC curve analysis revealed that the mean methylation level of the miR-657 promoter region could distinguish cancerous tissues from paired normal tissues of HCC patients (area under the curve: 0.847, P<0.001). Using 59.50% as the optimal cut-off, the sensitivity was 95.50% and the specificity was 70.01%. Conclusions: Methylation levels of the miR-657 promoter region were decreased in HCC patients and could be used as alternative and supplementary biomarkers for diagnosis of HCC.

Application of Taxol Nanomicelles with Lyp-1 Target in Targeted 805-813 Therapy of Colon Cancer.

As a new type of non-viral gene drug carrier, paclitaxel with Lyp-1 target has unique transmembrane ability due to its unique structure. In this paper, amino acids and surfactants are used to disperse SWCNTs in water, and non-covalent interactions are used to adsorb paclitaxel to the surface of SWCNTs. DSPE-PEG-Maleimide is then connected to NGR to achieve active targeting. To investigate the effect of NGR-SWCNTs-Paclitaxel on isolated cells, and to observe the antitumor effect of NGR-SWCNTs-Paclitaxel on S180 colon cancer mice in vivo, we provide theoretical and experimental basis for targeted cancer treatment. The luciferase activity test results showed that mi R-218 mimics had no significant effect on the intensity of the blank reporter plasmid group and p MIR-REPORT/UTR mutant luciferase activity, but in mi R-218 mimics and p MIR-REPORT/UTR Luciferase activity decreased after co-transfection of wild-type plasmids into cells. The validation results of the luciferase activity analysis system showed that mi R-218 was able to bind to Sp13'UTR. Overexpression of mi R-218 can significantly reduce the expression level of Sp1 protein but has no significant effect on Sp1 m RNA level, indicating that mi R-218 can target the regulation of Sp1 expression at the translation level.

Identification and Validation of a Novel Six-Gene Expression Signature for Predicting Hepatocellular Carcinoma Prognosis.

Background: Hepatocellular carcinoma (HCC) is a highly lethal disease. Effective prognostic tools to guide clinical decision-making for HCC patients are lacking. Objective: We aimed to establish a robust prognostic model based on differentially expressed genes (DEGs) in HCC. Methods: Using datasets from The Cancer Genome Atlas (TCGA), the Gene Expression Omnibus (GEO), and the International Genome Consortium (ICGC), DEGs between HCC tissues and adjacent normal tissues were identified. Using TCGA dataset as the training cohort, we applied the least absolute shrinkage and selection operator (LASSO) algorithm and multivariate Cox regression analyses to identify a multi-gene expression signature. Proportional hazard assumptions and multicollinearity among covariates were evaluated while building the model. The ICGC cohort was used for validation. The Pearson test was used to evaluate the correlation between tumor mutational burden and risk score. Through single-sample gene set enrichment analysis, we investigated the role of signature genes in the HCC microenvironment. Results: A total of 274 DEGs were identified, and a six-DEG prognostic model was developed. Patients were stratified into low- or high-risk groups based on risk scoring by the model. Kaplan-Meier analysis revealed significant differences in overall survival and progression-free interval. Through univariate and multivariate Cox analyses, the model proved to be an independent prognostic factor compared to other clinic-pathological parameters. Time-dependent receiver operating characteristic curve analysis revealed satisfactory prediction of overall survival, but not progression-free interval. Functional enrichment analysis showed that cancer-related pathways were enriched, while immune infiltration analyses differed between the two risk groups. The risk score did not correlate with levels of PD-1, PD-L1, CTLA4, or tumor mutational burden. Conclusions: We propose a six-gene expression signature that could help to determine HCC patient prognosis. These genes may serve as biomarkers in HCC and support personalized disease management.

Pulmonary multifocal arteriovenous malformations lead to ischemic stroke in young adults: a case report and literature review.

In recent years, the incidence of ischemic stroke in young people has gradually increased. The etiology and risk factors of young patients with ischemic stroke are complex, including migraine with aura, inherited thrombophilias, hyperhomocysteinemia, cardiovascular risk factors and malignancy. Pulmonary arteriovenous malformation (PAVM) is a rare cause of ischemic stroke in young people, which can easily be misdiagnosed and missed due to the lack of specific biomarkers. In this case report, we described a 17-year-old stroke patient who was admitted to hospital due to sudden speech failure accompanied by weakness of the right limb, and was diagnosed with cerebral embolism in the emergency department. After mechanical thrombectomy, the blood vessels were completely recanalized and the symptoms of nerve defects disappeared. Subsequent chest CT examination revealed pulmonary vascular malformations and further pulmonary angiography revealed multiple PAVMs in the lungs. Furthermore, spring coil embolization treatment was performed on 2 PAVMs with an inflow greater than 2 mm. Three months later, enhanced chest CT reexamination showed no recanalization of the malformed vessels at the pulmonary embolism site. Therefore, PAVM should be further not excluded for young stroke patients with unknown causes to avoid misdiagnosis and missed diagnosis, interventional embolization is the best treatment method after diagnosis.

The Preoperative Prognostic Nutritional Index in Hepatocellular Carcinoma After Curative Hepatectomy: A Retrospective Cohort Study and Meta-Analysis.

OBJECTIVE: Conflicting results existed about the role of prognostic nutritional index (PNI) for hepatocellular carcinoma (HCC) patients who received curative hepatectomy. The aim of this study is to identify the predictive capacity of PNI for survival after hepatectomy. METHODS: Preoperative PNI, neutrophil-to-lymphocyte ratio (NLR), tumor feature and clinical information of 187 patients with HCC from Sir Run Run Shaw hospital were evaluated. We also conducted a meta-analysis of seven cohort studies. RESULTS: Our study showed that HCC patients with a low PNI of <45 had a poor recurrence-free survival (RFS) rate (hazard ratio [HR] 1.762, 95% confidence interval [CI] 1.066-2.911, p = 0.027, respectively). The 5-year OS and RFS rates of the high PNI (≥45) vs low PNI (<45) were 76.7% vs 50.1% (p = 0.001) and 47.0% vs 28.9% (p = 0.001), respectively. In HCC TNM I patients (n = 144), a low PNI remained an independent prognostic factor of OS and RFS (HR 2.305, 95% CI 1.008-5.268, p = 0.048; HR 2.122, 95% CI 1.149-3.920, p = 0.016). The 5-year OS and RFS rates of the high PNI vs low PNI were 81.3% vs 62.4% (p = 0.041) and 53.4% vs 45.6% (p = 0.013), respectively. In the pooled analysis, the data showed that a low PNI was significantly associated with poor OS and RFS (HR 2.27, 95% CI 1.03-4.07, p < 0.001 and HR 1.68, 95% CI 1.45-1.94, p < 0.001, respectively). CONCLUSIONS: The preoperative PNI was an independent prognostic factor for OS and RFS rates in HCC patients who received hepatectomy.

Konjac glucomannan as a carrier material for time--temperature integrator.

Hardness, springiness and water retention of konjac glucomannan gel (g-KGM) as a novel carrier material for time-temperature integrator (TTI) in aseptic processing were determined and compared with those of sodium alginate gel (g-SA). Hardness of both g-KGM and g-SA increased with temperature: values of g-SA were significantly higher (p < 0.05) than those of g-KGM at all temperatures. No significant difference in springiness between g-KGM and g-SA from 40 °C to 90 °C and significant differences (p < 0.05) between 100 °C and 140 °C were found. Water retention property of g-KGM was lower than that of g-SA between 60 °C and 100 °C, but much higher between 100 °C and 140 °C. Heat transfer tests performed on g-KGM alone as well as on g-KGM as a carrier, embedded with TTI, α-amylase as an integrator, indicated that g-KGM was suitable for industrial ultrahigh temperature sterilization test.

Kinetic study of thermal inactivation of potato peroxidase during high-temperature short-time processing.

Thermal inactivation curves for peroxidase in potato extracts were determined in the range of 100 to 140°C for 10 to 100 sec. The capillary tube method was used to obtain isothermal conditions. The come-up time for the capillary tubes was accurately calculated by analysis method by which thermal inactivation kinetics of enzymes in relation to high temperature processing would be more easily detected. Heat inactivation of potato peroxidase followed first-order reaction kinetics and yielded a curved Arrhenius plot for the temperature dependence at high temperatures. Kinetics parameters, k and Ea, were calculated for potato peroxidase. At temperature range of 100-140°C, the activation energy of peroxidase was lower than that in the range of 78-84°C. It could be elucidated by the scheme of thermal inactivation pathway.

The Voice of Drug Consumers: Online Textual Review Analysis Using Structural Topic Model.

Many web-based pharmaceutical e-commerce platforms allow consumers to post open-ended textual reviews based on their purchase experiences. Understanding the true voice of consumers by analyzing such a large amount of user-generated content is of great significance to pharmaceutical manufacturers and e-commerce websites. The aim of this paper is to automatically extract hidden topics from web-based drug reviews using the structural topic model (STM) to examine consumers' concerns when they buy drugs online. The STM is a probabilistic extension of Latent Dirichlet Allocation (LDA), which allows the consolidation of document-level covariates. This innovation allows us to capture consumer dissatisfaction along with their dynamics over time. We extract 12 topics, and five of them are negative topics representing consumer dissatisfaction, whose appearances in the negative reviews are substantially higher than those in the positive reviews. We also come to the conclusion that the prevalence of these five negative topics has not decreased over time. Furthermore, our results reveal that the prevalence of price-related topics has decreased significantly in positive reviews, which indicates that low-price strategies are becoming less attractive to customers. To the best of our knowledge, our work is the first study using STM to analyze the unstructured textual data of drug reviews, which enhances the understanding of the aspects of drug consumer concerns and contributes to the research of pharmaceutical e-commerce literature.

CSF1R methylation is a key regulatory mechanism of tumor-associated macrophages in hepatocellular carcinoma.

Tumor-associated macrophages (TAMs) are important in tumor microenvironments and are closely associated with cancer occurrence, metastasis and progression. Colony stimulating factor 1 receptor (CSF1R) serves a crucial role in TAM formation. Whether CSF1R expression is regulated by DNA methylation in hepatocellular carcinoma (HCC) has not been fully elucidated. In the current study, HCC and adjacent non-cancerous tissue (ANT) samples were collected from 160 patients with HCC. CSF1R methylation levels were analyzed using a Mass ARRAY Analyzer to establish the potential impact of CSF1R methylation alternations on HCC clinicopathological characteristics. The mean methylation level of the CSF1R promoter (chr 5:149492491-149492958) was demonstrated to be significantly higher in ANTs compared with HCC tissues (65.3±7.5% vs. 57.3±14.4%, respectively; P<0.0001). CSF1R also exhibited decreased expression in HCC tissues compared with ANTs (P=0.0026). However, CSF1R expression was negatively correlated with CSF1R methylation levels in ANTs (r>0.4; P<0.0001). Further analysis indicated that patients with diabetes exhibited lower methylation levels in ANTs compared with HCC tissues (P=0.0062). Furthermore, CSF1R hypomethylation in ANTs was associated with a larger number of tumors (P=0.0332), larger tumor size (P=0.0494) and higher tumor grade (P=0.0244). Therefore, methylation alternation of the CSF1R promoter region analyzed in the present study was a key regulatory mechanism on CSF1R expression and ANT hypomethylation indicated poor clinicopathological characteristics of HCC. CSF1R may be a potential immunological therapeutic target for HCC.

Promoter aberrant methylation status of ADRA1A is associated with hepatocellular carcinoma.

The aim of our study was to explore the relationship between the methylation status of the alpha-1A adrenergic receptor (ADRA1A) gene and hepatocellular carcinoma (HCC). We combined our in-house data-set with the Cancer Genome Atlas (TCGA) data-set to screen and identify the methylation status and expression of adrenergic receptor (AR) genes in HCC. Immunohistochemistry and western blot were performed to assess the expression of ADRA1A in HCC cell lines and tissues. We further evaluated the methylation levels of the ADRA1A promoter region in 160 HCC patients using the Sequenom MassARRAY® platform and investigated the association between methylation of ADRA1A and clinical characteristics. The expression levels of ADRA1A mRNA and protein were significantly decreased in HCC tissues. Compared with that in paired normal tissues, the mean methylation level of the ADRA1A promoter region was significantly increased in tumour tissues from 160 HCC patients (25.2% vs. 17.0%, P < 0.0001). We found that a DNA methyltransferase inhibitor (decitabine) could increase the expression of ADRA1A mRNA in HCC cell lines. Moreover, hypermethylation of the ADRA1A gene in HCC samples was associated with clinical characteristics, including alcohol intake (P = 0.0097) and alpha-fetoprotein (P = 0.0411). Receiver operator characteristic (ROC) curve analysis demonstrated that the mean methylation levels of ADRA1A could discriminate between HCC tissues and adjacent non-cancerous tissues (AUC = 0.700, P < 0.0001). mRNA sequencing indicated that the main enriched pathways were pathways in cancer, cytokine-cytokine receptor interaction and metabolic pathways (P < 0.01). ADRA1A gene hypermethylation might contribute to HCC initiation and is a promising biomarker for the diagnosis of HCC.