Down-regulating of MFN2 promotes vascular calcification via regulating RAS-RAF-ERK1/2 pathway.

BACKGROUND: Vascular calcification (VC), as a prevalent feature of atherosclerosis (AS), is a life-threatening pathological change. Mitofusin 2 (MFN2) has been reported to be down-regulated and participate in the pathogenesis of AS. Here, we explored the feasible impacts of MFN2 on VC in AS. METHODS: Atherosclerotic lesion was evaluated by Oil Red O staining. The VC was detected by Alizarin Red S staining, ALP staining, and calcium content in vascular smooth muscle cells (VSMCs) or atherosclerotic mice. The chondrocyte differentiation of VSMCs was measured by Alcian blue staining. Western blotting and qRT-PCR were used to determine the protein and mRNA expression of associated molecules. Intermolecular interaction was measured by ChIP and dual luciferase assays. RESULTS: The expression of MFN2 and E2F1 was reduced in the aorta tissues of AS patients and mice. Silencing of MFN2 drove calcification in VSMCs and aortas of atherosclerotic mice as confirmed by up-regulating RUNX2, OPG levels, and down-regulating SM22α, α-SMA levels. The chondrocyte differentiation of VSMCs was accelerated by MFN2 knockdown through inducing the expression of Aggrecan, Collagen II, and SOX9. In addition, E2F1 promoted the transcription and expression of MFN2 in VSMCs. Overexpression of MFN2 or E2F1 suppressed ox-LDL-induced VSMC calcification. Finally, MFN2 depletion enhanced VSMC calcification via activating RAS-RAF-ERK1/2 pathway. CONCLUSION: Our results suggest that silencing of MFN2 drives VC via activating RAS-RAF-ERK1/2 pathway in the progression of AS, thus MFN2 may be a therapeutic target for AS.

Novel model combining contrast-enhanced ultrasound with serology predicts hepatocellular carcinoma recurrence after hepatectomy.

BACKGROUND: Surgery is the primary curative option in patients with hepatocellular carcinoma (HCC). However, recurrence within 2 years is observed in 30%-50% of patients, being a major cause of mortality. AIM: To construct and verify a non-invasive prediction model combining contrast-enhanced ultrasound (CEUS) with serology biomarkers to predict the early recurrence of HCC. METHODS: Records of 744 consecutive patients undergoing first-line curative surgery for HCC in one institution from 2016-2018 were reviewed, and 292 local patients were selected for analysis. General characteristics including gender and age, CEUS liver imaging reporting and data system (LIRADS) parameters including wash-in time, wash-in type, wash-out time, and wash-out type, and serology biomarkers including alanine aminotransferase, aspartate aminotransferase, platelets, and alpha-fetoprotein (AFP) were collected. Univariate analysis and multivariate Cox proportional hazards regression model were used to evaluate the independent prognostic factors for tumor recurrence. Then a nomogram called CEUS model was constructed. The CEUS model was then used to predict recurrence at 6 mo, 12 mo, and 24 mo, the cut-off value was calculate by X-tile, and each C-index was calculated. Then Kaplan-Meier curve was compared by log-rank test. The calibration curves of each time were depicted. RESULTS: A nomogram predicting early recurrence (ER), named CEUS model, was formulated based on the results of the multivariate Cox regression analysis. This nomogram incorporated tumor diameter, preoperative AFP level, and LIRADS, and the hazard ratio was 1.123 (95% confidence interval [CI]: 1.041-1.211), 1.547 (95%CI: 1.245-1.922), and 1.428 (95%CI: 1.059-1.925), respectively. The cut-off value at 6 mo, 12 mo, and 24 mo was 100, 80, and 50, and the C-index was 0.748 (95%CI: 0.683-0.813), 0.762 (95%CI: 0.704-0.820), and 0.762 (95%CI: 0.706-0.819), respectively. The model showed satisfactory results, and the calibration at 6 mo was desirable; however, the calibration at 12 and 24 mo should be improved. CONCLUSION: The CEUS model enables the well-calibrated individualized prediction of ER before surgery and may represent a novel tool for biomarker research and individual counseling.