Cell-free DNA methylation markers for differential diagnosis of hepatocellular carcinoma.

BACKGROUND: Aberrant DNA methylation may offer opportunities in revolutionizing cancer screening and diagnosis. We sought to identify a non-invasive DNA methylation-based screening approach using cell-free DNA (cfDNA) for early detection of hepatocellular carcinoma (HCC). METHODS: Differentially, DNA methylation blocks were determined by comparing methylation profiles of biopsy-proven HCC, liver cirrhosis, and normal tissue samples with high throughput DNA bisulfite sequencing. A multi-layer HCC screening model was subsequently constructed based on tissue-derived differentially methylated blocks (DMBs). This model was tested in a cohort consisting of 120 HCC, 92 liver cirrhotic, and 290 healthy plasma samples including 65 hepatitis B surface antigen-seropositive (HBsAg+) samples, independently validated in a cohort consisting of 67 HCC, 111 liver cirrhotic, and 242 healthy plasma samples including 56 HBsAg+ samples. RESULTS: Based on methylation profiling of tissue samples, 2321 DMBs were identified, which were subsequently used to construct a cfDNA-based HCC screening model, achieved a sensitivity of 86% and specificity of 98% in the training cohort and a sensitivity of 84% and specificity of 96% in the independent validation cohort. This model obtained a sensitivity of 76% in 37 early-stage HCC (Barcelona clinical liver cancer [BCLC] stage 0-A) patients. The screening model can effectively discriminate HCC patients from non-HCC controls, including liver cirrhotic patients, asymptomatic HBsAg+ and healthy individuals, achieving an AUC of 0.957(95% CI 0.939-0.975), whereas serum α-fetoprotein (AFP) only achieved an AUC of 0.803 (95% CI 0.758-0.847). Besides detecting patients with early-stage HCC from non-HCC controls, this model showed high capacity for distinguishing early-stage HCC from a high risk population (AUC=0.934; 95% CI 0.905-0.963), also significantly outperforming AFP. Furthermore, our model also showed superior performance in distinguishing HCC with normal AFP (< 20ng ml-1) from high risk population (AUC=0.93; 95% CI 0.892-0.969). CONCLUSIONS: We have developed a sensitive blood-based non-invasive HCC screening model which can effectively distinguish early-stage HCC patients from high risk population and demonstrated its performance through an independent validation cohort. TRIAL REGISTRATION: The study was approved by the ethic committee of The Second Xiangya Hospital of Central South University (KYLL2018072) and Chongqing University Cancer Hospital (2019167). The study is registered at ClinicalTrials.gov(# NCT04383353 ).

Construction and Analysis of a Long Non-Coding RNA-Associated Competing Endogenous RNA Network Identified Potential Prognostic Biomarkers in Luminal Breast Cancer.

PURPOSE: To construct a competing endogenous RNA (ceRNA) topology network of RNA-seq data and micro RNA-seq (miRNA-seq) data to identify key prognostic long non-coding RNA (lncRNAs) in luminal breast cancer, and validate the results by human luminal breast cancer samples. MATERIALS AND METHODS: The RNA-seq data and miRNA-seq data of luminal A breast cancer in the The Cancer Genome Atlas (TCGA) database were downloaded and compared with those in the miRcode database to obtain lncRNA-miRNA relationship pairs. Final target genes were predicted by all three databases (miRDB, miRTarBase, and TargetScan), thereby obtaining the miRNA-messenger RNA (miRNA-mRNA) relationship pairs and a ceRNA topology network was constructed, then mRNA enrichment analysis, ceRNA topological and stability analysis, univariate and multivariate Cox regression analysis were performed. Overall survival (OS) was evaluated and the key prognostic RNAs were identified. The expression difference between normal and tumor, as well as the correlation of high expression in tumor with pathological parameters (Ki-67, Grade, tumor diameter) were validated by human breast cancer specimens. RESULTS: A ceRNA topology network was constructed and six lncRNAs were finally identified (The higher expression of PART1, IGF2.AS, WT1.AS, OIP5.AS1, and SLC25A5.AS1 was associated with poor prognosis while AL035706.1 was adverse) and the poor prognostic ones were higher expressed in tumor tissue and correlated with a higher Ki-67 (>10%), tumor grades (II, III) and tumor diameters (>1.5 cm). Using six lncRNAs, we constructed a prognostic model, which performed well for the classification of prognosis in the module. CONCLUSION: We identified and verified six biomarkers (OS-predicting) in luminal breast cancer, which significantly enriched the prediction and potential targets of this subtype.

Individualized comprehensive treatment for lung adenocarcinoma with multiple skin and bilateral breast metastasis: A case report. / å¤šæ¬¡çš®è‚¤å’ŒåŒä¾§ä¹³è ºè½¬ç§»çš„è‚ºè ºç™Œä¸ªä½“åŒ–ç»¼åˆæ²»ç–—1例.

Lung adenocarcinoma is a malignant tumor that is prone to distant metastasis. Common metastatic sites are brain, adrenal gland, liver, bone, and so on. Skin soft tissue metastasis is unusual, and breast metastasis is even rarer. This case is a middle-aged female patient who had experienced multi-line treatments for upper limbs, abdominal skin, and bilateral breast tissue metastases.The patient's multiple metastases were susceptible to radiation therapy.Reviewing the entire treatment process of this patient can find that the rational use of individualized comprehensive treatment methods and appropriate timing of genetic testing are very important for patients with lung adenocarcinoma to prolong their survival time and improve their quality of life.

Clinical and prognostic features of 512 cases of cholangiocarcinoma. / 512例胆管细胞癌的临床及预后特点.

OBJECTIVES: Cholangiocarcinoma (CCA) is an aggressive malignant tumor with a poor overall prognosis. Given that CCA is often diagnosed at the late stage, the current treatments are less effective for most local advanced patients leading to high CCA mortality. This study aims to explore the clinical characteristics and prognostic factors affecting the occurrence and development of CCA and to provide potential methods for early diagnosis and clinical treatment of CCA. METHODS: We retrospectively analyzed the medical records of 512 patients with CCA who had been diagnosed by pathology and had completely clinical data in the Second Xiangya Hospital of Central South University in the past 16 years. The clinical features and prognosis related factors that affect the occurrence and development of CCA were investigated. Survival curves were plotted by the Kaplan-Meier method. P-values were calculated by log-rank for univariate analysis, and multivariate Cox regression was used to analyze multivariate analysis of meaningful variables. RESULTS: The incidence of CCA among ≤60 years old people was higher than of that >60 years old one (61.13% vs 38.87%), and was greater in men than women (52.5% vs 47.5%). Carbohydrate antigen 19-9 (CA19-9) level ≥35 µg/L accounted for 66.21%. The single tumor accounted for 86.91%, and patients in pathological stage III and IV accounted for 49.22% and 17.58%, respectively. Univariate analysis showed that ALB, ALP, CA19-9, and other factors were relevant to the prognosis. The results of multivariate analysis showed that ALP, CA19-9, tmaximum tumor diameter, and other factors were significant prognostic predictors. CONCLUSIONS: The incidence of CCA is higher in ≤60 years old people, and the stage is later at the initial diagnosis. CA19-9 level is a sensitive laboratory indicator. ALP, CA19-9, maximum tumor diameter, merged tumor, cirrhosis, and TNM stage are independent prognostic factors for CCA.