A Seven-microRNA Expression Signature Predicts Survival in Hepatocellular Carcinoma

<div><p>Hepatocellular carcinoma (HCC) is the fifth common cancer. The differential expression of microRNAs (miRNAs) has been associated with the prognosis of various cancers. However, limited information is available regarding genome-wide miRNA expression profiles in HCC to generate a tumor-specific miRNA signature of prognostic values. In this study, the miRNA profiles in 327 HCC patients, including 327 tumor and 43 adjacent non-tumor tissues, from The Cancer Genome Atlas (TCGA) Liver hepatocellular carcinoma (LIHC) were analyzed. The associations of the differentially expressed miRNAs with patient survival and other clinical characteristics were examined with t-test and Cox proportional regression model. Finally, a tumor-specific miRNA signature was generated and examined with Kaplan–Meier survival, univariate\multivariate Cox regression analyses and KEGG pathway analysis. Results showed that a total of 207 miRNAs were found differentially expressed between tumor and adjacent non-tumor HCC tissues. 78 of them were also discriminatively expressed with gender, race, tumor grade and AJCC tumor stage. Seven miRNAs were significantly associated with survival (P value <0.001). Among the seven significant miRNAs, six (hsa-mir-326, hsa-mir-3677, hsa-mir-511-1, hsa-mir-511-2, hsa-mir-9-1, and hsa-mir-9-2) were negatively associated with overall survival (OS), while the remaining one (hsa-mir-30d) was positively correlated. A tumor-specific 7-miRNAs signature was generated and validated as an independent prognostic predictor. Collectively, we have identified and validated an independent prognostic model based on the expression of seven miRNAs, which can be used to assess patients’ survival. Additional work is needed to translate our model into clinical practice.</p></div

Heatmap of KEGG pathways enriched in seven miRNA target genes.

<p>The isoforms of 7-miRNA signature were involved in multiple pathways, especially cancer-specific pathways. (DIANA-mirpath computes log₁₀ P-values).</p

Kaplan-Meier survival curves for 7-miRNA signature.

<p>Six miRNA (hsa-mir-326, hsa-mir-3677, hsa-mir-511-1, hsa-mir-511-2, hsa-mir-9-1, and hsa-mir-9-2) were negatively associated with OS and one (hsa-mir-30d) was positively correlated. (Red line: overexpressed, Blue line: under-expressed, Horizontal axis: overall survival time, Vertical axis: survival function).</p

Fifty differentially expressed miRNAs (Absolute fold changes >3).

<p>The expression levels of the significant miRNAs in hepatocellular cell carcinoma (HCC) are compared to those of adjacent non-tumor liver tissue, as fold change (tumor vs. non-tumor).</p

Heatmap of 207 differentially expressed miRNAs in tumor/non-tumor.

<p>The miRNA expression levels are mean-centered across samples, and an unsupervised hierarchical clustering with complete linkage is carried out by Cluster 3.0.</p

Clinical characteristics of patients with hepatocellular cell carcinoma.

<p>AJCC: American Joint Committee on Cancer; NA: Not Available; Risk factors: Alcohol consumption, Hepatitis B/C, Hemochromatosis, Nonalcoholic Fatty Liver Disease, Alpha-1 Antitrypsin Deficiency</p><p>^a Statistical significant results (in bold)</p><p>Clinical characteristics of patients with hepatocellular cell carcinoma.</p

Differentially expressed miRNAs from Tumor/Non-tumor according to clinical parameters.

<p>AJCC American Joint Committee on Cancer; Tumor grade: neoplasm histologic grade; vs. versus.</p><p>Differentially expressed miRNAs from Tumor/Non-tumor according to clinical parameters.</p

Kaplan–Meier survival curves for hepatocellular cell carcinoma patients.

<p>The 327 hepatocellular cell carcinoma patients are compared in two groups according to: a 7-miRNA signature (1 high risk vs. 2 low risk); b Tumor status (1 Tumor free vs. 2 With tumor); c AJCC pathological (1 I + II vs. 2 stage III + IV); and d AJCC T stage (1 T1+T2 vs. 2 T3+T4). Log Rank (Mantel-Cox) P value are 0.000, 0.001, 0.042 and 0.002, respectively. (Blue line: group 1, Green line: group 2, Horizontal axis: overall survival time, Vertical axis: survival function).</p

A combined endoscopic and robotic approach for Mirizzi syndrome: the short- and long-term outcomes

Aims: We present the technique of a combined endoscopic and robotic approach for Mirizzi syndrome (MS) and report the short- and long-term results.Methods: Between July 2012 and August 2020, all patients with suspected MS underwent endoscopic retrograde cholangiopancreatography (ERCP) for diagnostic confirmation and placement of the biliary stent. Subtotal cholecystectomy was then performed with the assistance of a surgical robot. The common bile duct was closed with a cuff of the gallbladder over a biliary stent. ERCP was repeated 6-8 weeks after surgery to remove the biliary stent and confirm the patency of common bile duct. The operative outcomes and long-term results were prospectively collected.Results: Twenty-two patients (10 males and 12 females) were included in the study. All patients underwent the planned robotic subtotal cholecystectomy and pre- and postoperative ERCP. The median age was 65.5 years (range 16-89 years). The median operative time was 212.5 min (range 125-510 min), and the median blood loss was 35 mL (range 7-700 mL). The median postoperative hospital stay was four days (range 3-15 days). Four patients (18.2%) developed postoperative complications including two intra-abdominal collections (9.1%), one wound infection (4.5%), and one atrial fibrillation with pneumonia (4.5%). The last patient also represented the only operative mortality. The median follow-up was 45.6 months; none developed recurrent cholangitis or jaundice, but one patient (4.5%) had a recurrent common bile duct stone which was successfully removed by ERCP.Conclusion: The proposed combined endoscopic and robotic approach can provide favorable short- and long-term outcomes for patients with MS