Thermal stability and interfacial rheology of whey protein-corn oil emulsions: The effect of l-arginine fortification.

The effect of l-arginine (Arg) on the thermal stability of whey protein-corn oil emulsions was investigated to determine its role in improving emulsion stability. The results indicated that with an increase in Arg concentration, the emulsion stability index, emulsification activity index, and absolute ζ-potential increased initially and decreased after high-temperature sterilization. However, the mean particle size, apparent viscosity, creaming indices, and dynamic interfacial pressure of the emulsions first decreased and then increased, and the performance of samples that only showed an increase in pH could also improve the emulsification stability. These results clarify the mechanism by which Arg increases the thermal stability of emulsions.

Evaluation of left ventricular myocardial work in patients with hyperthyroidism with different heart rates with noninvasive pressure-strain loop based on two-dimensional speck tracking imaging.

Background: We investigated the application value of no-invasive myocardial work in evaluating left ventricular (LV) function in patients with hyperthyroidism. Methods: Sixty-five patients with an initial hyperthyroidism diagnosis were sorted into tachycardia (group TH1, n=31) and without tachycardia (group TH2, n=34) groups. Thirty healthy participants served as the control group (group CON). LV strain parameters and LV myocardial work parameters were evaluated at rest. Each parameter's value in identifying myocardial damage was analyzed using receiver operating characteristic curves. The correlation of myocardial work parameters with global longitudinal strain (GLS), longitudinal peak strain dispersion (normalized by heart rate, PSDN), and systolic blood pressure (SBP) was analyzed. Results: There was no difference in classic echocardiographic parameters between the groups. Compared with that in group CON, GLS decreased in groups TH1 and TH2 (TH1 17.99%±2.21% and TH2: 19.00%±2.85% vs. 20.27%±1.49%; both P<0.05); there was no significant difference between groups TH1 and TH2. PSDN increased in groups TH1 and TH2 (TH1 73.13±19.51 ms and TH2 55.06±17.03 vs. 44.13±8.65 ms; both P<0.05); it was higher in group TH1 than in group TH2 (P<0.05). Myocardial global work efficiency (GWE) decreased in groups TH1 and TH2 {TH1 95% [interquartile range (IQR), 94-95%] and TH2 96% (IQR, 95-97%) vs. 97% (IQR, 96-97%); both P<0.05}; it was lower in group TH1 than in group TH2 (P<0.05). Global constructive work (GCW) decreased in group TH1 (1,865.29±284.13 vs. 2,030.33±252.52 mmHg%; P<0.05), but was not different from that in group TH2; there was no difference between groups TH2 and CON. Global wasted work (GWW) increased in groups TH1 and TH2 [TH1 83.00 (IQR, 74.00-97.00) mmHg% and TH2 69.50 (IQR, 51.25-84.25) vs. 50.50 (IQR, 40.75-65.25) mmHg%; both P<0.05]; it was higher in group TH1 than in group TH2 (P<0.05). The area under the GWE curve was the largest (area under the curve =0.835), and the optimal cutoff point was 96.5%, with a sensitivity of 0.83 and a specificity of 0.70. GWE and GCW were positively correlated with GLS and negatively correlated with PSDN. GWW was negatively correlated with GLS and positively correlated with PSDN. In group CON, GCW and GWW were positively correlated with SBP; GWE was not correlated with SBP. In groups TH1 and TH2, GCW was positively correlated with SBP, but not with GWW or GWE. Conclusions: Hyperthyroidism can significantly decrease the GWE and increase GWW of the left ventricle. This change is more pronounced in patients with tachycardia. Myocardial work could be a novel method for the evaluation of LV myocardial function in patients with hyperthyroidism.

Prediction of Surgical Approach in Mitral Valve Disease by XGBoost Algorithm Based on Echocardiographic Features.

In this study, we aimed to develop a prediction model to assist surgeons in choosing an appropriate surgical approach for mitral valve disease patients. We retrospectively analyzed a total of 143 patients who underwent surgery for mitral valve disease. The XGBoost algorithm was used to establish a predictive model to decide a surgical approach (mitral valve repair or replacement) based on the echocardiographic features of the mitral valve apparatus, such as leaflets, the annulus, and sub-valvular structures. The results showed that the accuracy of the predictive model was 81.09% in predicting the appropriate surgical approach based on the patient's preoperative echocardiography. The result of the predictive model was superior to the traditional complexity score (81.09% vs. 75%). Additionally, the predictive model showed that the three main factors affecting the choice of surgical approach were leaflet restriction, calcification of the leaflet, and perforation or cleft of the leaflet. We developed a novel predictive model using the XGBoost algorithm based on echocardiographic features to assist surgeons in choosing an appropriate surgical approach for patients with mitral valve disease.

Construction and validation of a progression prediction model for locally advanced rectal cancer patients received neoadjuvant chemoradiotherapy followed by total mesorectal excision based on machine learning.

Background: We attempted to develop a progression prediction model for local advanced rectal cancer(LARC) patients who received preoperative neoadjuvant chemoradiotherapy(NCRT) and operative treatment to identify high-risk patients in advance. Methods: Data from 272 LARC patients who received NCRT and total mesorectal excision(TME) from 2011 to 2018 at the Fourth Hospital of Hebei Medical University were collected. Data from 161 patients with rectal cancer (each sample with one target variable (progression) and 145 characteristic variables) were included. One Hot Encoding was applied to numerically represent some characteristics. The K-Nearest Neighbor (KNN) filling method was used to determine the missing values, and SmoteTomek comprehensive sampling was used to solve the data imbalance. Eventually, data from 135 patients with 45 characteristic clinical variables were obtained. Random forest, decision tree, support vector machine (SVM), and XGBoost were used to predict whether patients with rectal cancer will exhibit progression. LASSO regression was used to further filter the variables and narrow down the list of variables using a Venn diagram. Eventually, the prediction model was constructed by multivariate logistic regression, and the performance of the model was confirmed in the validation set. Results: Eventually, data from 135 patients including 45 clinical characteristic variables were included in the study. Data were randomly divided in an 8:2 ratio into a data set and a validation set, respectively. Area Under Curve (AUC) values of 0.72 for the decision tree, 0.97 for the random forest, 0.89 for SVM, and 0.94 for XGBoost were obtained from the data set. Similar results were obtained from the validation set. Twenty-three variables were obtained from LASSO regression, and eight variables were obtained by considering the intersection of the variables obtained using the previous four machine learning methods. Furthermore, a multivariate logistic regression model was constructed using the data set; the ROC indicated its good performance. The ROC curve also verified the good predictive performance in the validation set. Conclusions: We constructed a logistic regression model with good predictive performance, which allowed us to accurately predict whether patients who received NCRT and TME will exhibit disease progression.

Early assessment of subclinical myocardial injury in systemic lupus erythematosus by two-dimensional longitudinal layer speckle tracking imaging.

Background: To investigate the feasibility of quantitatively assessing left ventricular function and synchronization and diagnose subclinical myocardial injury in patients with systemic lupus erythematosus (SLE) using two-dimensional (2D) longitudinal layer speckle tracking imaging (STI). Methods: This was a single-center prospective study. A total of 69 patients with SLE were included in the case group and further divided into 2 subgroups, a nonactive and an active group, according to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) 2000 scoring standard. We selected 30 healthy volunteers as the control group. The global longitudinal strain (GLSglobal), global endocardial longitudinal strain (GLSendo), global epicardial longitudinal strain (GLSepi), and peak strain dispersion (PSD) were obtained. The transmural gradient of longitudinal strain (TGLS) was calculated for the difference in strains between the inner and outer membranes. Results: (I) Compared with the control group, decreased speckle strain parameters and elevated PSD were observed in patients with SLE (GLSglobal: -18.80%±2.41% vs. -21.19%±2.16%, GLSendo: -21.15%±2.47% vs. -24.09±2.49%; GLSepi: -16.58%±2.39% vs. -18.50±1.77%; TGLS: -4.56%±1.24% vs. -5.59%±1.39%; and PSD: 36.61±10.85 vs. 30.00±8.54 ms). More severely impaired layer strains were observed in active-stage patients. Compared with the nonactive group, GLSendo, GLSglobal, GLSepi, TGLS, complement C3, and complement C4 were decreased in the active group, while SLEDAI, erythrocyte sedimentation rate (ESR), and high-sensitivity C-reactive protein (Hs-CRP) were elevated. (II) Receiver operating characteristic (ROC) analysis demonstrated that subendocardial myocardial longitudinal strain was the most powerful tool for detecting myocardial insufficiency early in patients with SLE [area under the curve (AUC) =0.809], especially in patients in the active stage (AUC =0.734), and the optimal cut-off point was -21.35%, with a sensitivity of 71.9% and a specificity of 62.2%. (III) Correlation analysis revealed that GLSendo was moderately correlated with PSD, SLEDAI, ERS, Hs-CRP, and complement C3 (correlation coefficients: 0.535, 0.428, 0.659, 0.559, and -0.440, respectively). Conclusions: Subclinical myocardial injury in patients with SLE can be assessed early using 2D longitudinal STI, and the injury is more obvious in active-stage patients. Endocardial longitudinal strain is a more sensitive index than epicardial longitudinal strain for the early detection of subclinical myocardial injury in patients with SLE, which is a potentially valuable clinical tool to assist in the early detection of myocardial damage.

Ethanol organosolv lignin from different agricultural residues: Toward basic structural units and antioxidant activity.

The growing interest in the substitution of synthetic food antioxidants by natural ones has fostered research on vegetable sources and the screening of raw materials for identifying new antioxidants. Special attention is focused on their extraction from inexpensive or residual sources from agricultural industries. Herein, the antioxidant activities of lignin obtained from 4 residual sources were investigated. The obtained lignin samples were characterized by different analytical techniques evaluating their chemical structure, phenolic content, thermal behavior and molecular weight. The antioxidant activity of the analyzed lignins was evaluated by the DPPH assay, the radical ABTS assay, and trivalent iron reduction method. It was found t that lignin antioxidants could scavenge free radicals and reduce oxidants. The high correlation between antioxidant capacity and its total phenol content indicated that phenolic hydroxyl groups were the main contributors to these lignins' antioxidant activity.

Exosomal S100A4 derived from highly metastatic hepatocellular carcinoma cells promotes metastasis by activating STAT3.

Intercellular cross-talk plays important roles in cancer progression and metastasis. Yet how these cancer cells interact with each other is still largely unknown. Exosomes released by tumor cells have been proved to be effective cell-to-cell signal mediators. We explored the functional roles of exosomes in metastasis and the potential prognostic values for hepatocellular carcinoma (HCC). Exosomes were extracted from HCC cells of different metastatic potentials. The metastatic effects of exosomes derived from highly metastatic HCC cells (HMH) were evaluated both in vitro and in vivo. Exosomal proteins were identified with iTRAQ mass spectrum and verified in cell lines, xenograft tumor samples, and functional analyses. Exosomes released by HMH significantly enhanced the in vitro invasion and in vivo metastasis of low metastatic HCC cells (LMH). S100 calcium-binding protein A4 (S100A4) was identified as a functional factor in exosomes derived from HMH. S100A4rich exosomes significantly promoted tumor metastasis both in vitro and in vivo compared with S100A4low exosomes or controls. Moreover, exosomal S100A4 could induce expression of osteopontin (OPN), along with other tumor metastasis/stemness-related genes. Exosomal S100A4 activated OPN transcription via STAT3 phosphorylation. HCC patients with high exosomal S100A4 in plasma also had a poorer prognosis. In conclusion, exosomes from HMH could promote the metastatic potential of LMH, and exosomal S100A4 is a key enhancer for HCC metastasis, activating STAT3 phosphorylation and up-regulating OPN expression. This suggested exosomal S100A4 to be a novel prognostic marker and therapeutic target for HCC metastasis.

Screen the Effective Components of Lycopodii herba on Rheumatoid Arthritis with the Aid of Spectrum-Effect Relationship and Uncover its Potential Mechanism.

Lycopodii herba (SJC), a traditional Chinese medicine, has the effect of dispelling wind and eliminating dampness (a therapeutic principle and method of traditional Chinese medicine for rheumatoid arthritis), relaxing tendon and activating collaterals. However, the major effective components and its therapeutic mechanism were unclear. In this study, different SJC samples with slightly different compositions were prepared by extracting with different concentrations of ethanol. Then, the therapeutic effects on rheumatoid arthritis (RA) of different SJC samples were evaluated. Finally, the spectrum-effect relationship between UPLC-Q-TOF/MS fingerprints and the effect of RA was explored to screen the effective components. Western blotting was used to study the potential mechanism. The volume of hind paw and the level of RF, TNF-α, and IL-1ß were lower after administrating with different SJC samples, compared with the model group. Histopathological findings also confirmed that SJC could relieve the symptoms of RA. Combined with identification of the components in plasm from SJC, lycojaponicumin C, des-N-methyl-α-obscurine, 8ß-acetoxy-12ß-hydroxy-lycopodine or 8ß-acetoxy-11α-hydroxy-lycopodine or 8ß-hydroxy-11α-acetoxylycopodine were considered to be the major effective components. The mechanism may be related to AChE/NF-κB signaling pathway. This work provides a general method to screen the potential effective components of herb medicines and would be benefit to understand the mechanism of SJC for the treatment of RA.

Tailor-made ternary nanopolyplexes of thiolated trimethylated chitosan with pDNA and folate conjugated cis-aconitic amide-polyethylenimine for efficient gene delivery.

To overcome the different extra-/intracellular barriers in gene delivery, tumor-targeted and pH/redox-responsive ternary polyplexes with charge-conversional properties were prepared through a modular self-assembly strategy. Firstly, the thiolated trimethylated chitosan (TMC-SH) was synthesized to crosslink and condense pDNA through electrostatic interaction and disulfide formation, which obtained the TMC-SS/pDNA binary polyplexes with redox-responsive gene release. To further endow the binary polyplexes with tumor targeting and endo/lysosomal pH-triggered charge-reversal properties, a folate conjugated cis-aconitic amide-polyethylenimine (FA-PEI-AcO) was synthesized to shield the positive TMC-SS/pDNA, generating the FA-PEI-AcO/TMC-SS/pDNA ternary polyplexes with a size of ~190 nm and negative surface-charges. The ζ-potential of the polyplexes was stable at physiological pH and increased rapidly from -14 mV to + 20 mV at pH 5.5 (endo/lysosomal pH) due to the breakages of acid-liable amide bonds and the subsequent de-shielding of FA-PEI-AcO layers, which might benefit the endo/lysosomal escape of the polyplexes. Afterward, the polyplexes could redox-responsively release gene at higher intracellular concentrations of glutathione. By taking advantage of such multi-responses, significantly enhanced transfection efficiency was achieved in vitro in Hela cells for the ternary polyplexes. These results suggested that the newly developed polyplexes had potential application for gene delivery.

Genome-Wide Association Study Identifies a Genetic Prediction Model for Postoperative Survival in Patients with Hepatocellular Carcinoma.

BACKGROUND As an important aspect of tumor heterogeneity, genetic variation may influence susceptibility and prognosis in different types of cancer. By exploring the prognostic value of genetic variation, this study aimed to establish a model for predicting postoperative survival and assessing the impact of variation on clinical outcomes in patients with hepatocellular carcinoma (HCC). MATERIAL AND METHODS A genome-wide association study of 367 patients with HCC was conducted to identify single nucleotide polymorphisms (SNPs) associated with prognosis. Identified predictors were further evaluated in 758 patients. Two prognostic models were established using Cox proportional hazards regression and Nomogram strategy, and validated in another 316 patients. The effect of the SNP rs2431 was analyzed in detail. RESULTS A prognostic model including 5 SNPs (rs10893585, rs2431, rs34675408, rs6078460, and rs6766361) was established and exhibited high predictive accuracy for HCC prognosis. The panel combined with tumor node metastasis (TNM) stage resulted in a significantly higher c-index (0.723) than the individual c-index values. Stratified by the Nomogram prediction model, the median overall survival for the low-risk and high-risk groups were 100.1 versus 30.8 months (P<0.001) in the training set and 82.2 versus 22.5 months (P<0.001) in the validation set. A closer examination of rs2431 revealed that it may regulate the expression of FNDC3B by disrupting a microRNA-binding site. CONCLUSIONS This study established prediction models based on genetic factors alone or in combination with TNM stage for postoperative survival in patients with HCC, and identified FNDC3B as a potential therapeutic target for combating HCC metastasis.

Cideb controls sterol-regulated ER export of SREBP/SCAP by promoting cargo loading at ER exit sites.

SREBPs are master regulators of lipid homeostasis and undergo sterol-regulated export from ER to Golgi apparatus for processing and activation via COPII-coated vesicles. While COPII recognizes SREBP through its escort protein SCAP, factor(s) specifically promoting SREBP/SCAP loading to the COPII machinery remains unknown. Here, we show that the ER/lipid droplet-associated protein Cideb selectively promotes the loading of SREBP/SCAP into COPII vesicles. Sterol deprivation releases SCAP from Insig and enhances ER export of SREBP/SCAP by inducing SCAP-Cideb interaction, thereby modulating sterol sensitivity. Moreover, Cideb binds to the guanine nucleotide exchange factor Sec12 to enrich SCAP/SREBP at ER exit sites, where assembling of COPII complex initiates. Loss of Cideb inhibits the cargo loading of SREBP/SCAP, reduces SREBP activation, and alleviates diet-induced hepatic steatosis. Our data point to a linchpin role of Cideb in regulated ER export of SREBP and lipid homeostasis.

Disruption of tumour-associated macrophage trafficking by the osteopontin-induced colony-stimulating factor-1 signalling sensitises hepatocellular carcinoma to anti-PD-L1 blockade.

OBJECTIVE: In the tumour microenvironment, critical drivers of immune escape include the oncogenic activity of the tumour cell-intrinsic osteopontin (OPN), the expression of programmed death ligand 1 (PD-L1) and the expansion of tumour-associated macrophages (TAMs). We investigated the feasibility of targeting these pathways as a therapeutic option in hepatocellular carcinoma (HCC) mouse models. DESIGN: We analysed the number of tumour-infiltrating immune cells and the inflammatory immune profiles in chemically induced liver tumour isolated from wild-type and OPNknockout (KO) mice. In vitro cell cocultures were further conducted to investigate the crosstalk between TAMs and HCC cells mediated by OPN, colony stimulating factor-1 (CSF1) and CSF1 receptor (CSF1R). The in vivo efficacy of anti-PD-L1 and CSF1/CSF1R inhibition was evaluated in OPN overexpressing subcutaneous or orthotopic mouse model of HCC. RESULTS: The numbers of TAMs, as well as the expression levels of M2 macrophage markers and PD-L1 were significantly decreased, but the levels of cytokines produced by T-helper 1 (Th1) cells were upregulated in tumour tissues from OPN KO mice compared with that from the controls. In addition, we observed a positive association between the OPN and PD-L1 expression, and OPN expression and TAM infiltration in tumour tissues from patients with HCC. We further demonstrated that OPN facilitates chemotactic migration, and alternative activation of macrophages, and promotes the PD-L1 expression in HCC via activation of the CSF1-CSF1R pathway in macrophages. Combining anti-PD-L1 and CSF1R inhibition elicited potent antitumour activity and prolonged survival of OPNhigh tumour-bearing mice. Histological, flow cytometric and ELISA revealed increased CD8+ T cell infiltration, reduced TAMs and enhanced Th1/Th2 cytokine balance in multiple mouse models of HCC. CONCLUSIONS: OPN/CSF1/CSF1R axis plays a critical role in the immunosuppressive nature of the HCC microenvironment. Blocking CSF1/CSF1R prevents TAM trafficking and thereby enhances the efficacy of immune checkpoint inhibitors for the treatment of HCC.

Mutated EPHA2 is a target for combating lymphatic metastasis in intrahepatic cholangiocarcinoma.

Exploring the genetic aberrations favoring metastasis is important for understanding and developing novel strategies to combat cancer metastasis. It remains lack of effective treatment for the dismal prognosis of intrahepatic cholangiocarcinoma (ICC). Here, we aimed to study genetic alternations during lymph node metastasis of ICC and investigate potential mechanisms and clinical strategy focused on mutations. We performed whole-exome sequencing and transcriptome sequencing on samples from 30 ICC patients, including lymph node metastases from five of the patients. We identified the alterations of genetic pattern related to lymph node metastases of ICC. EPHA2, a member of the tyrosine kinase family, was found to be frequently mutated in ICC. Correlation analysis indicated that EPHA2 mutations were closely associated with lymph node metastasis of ICC. In vitro and in vivo experiments revealed that EPHA2 mutations could lead to ligand independent phosphorylation of Ser897, and promote lymphatic metastasis of ICC, in which NOTCH1 signaling pathway played an important role. In both in vitro assays and patient-derived xenografts, an inhibitor of Ser897 phosphorylation effectively suppressed the metastasis of ICC with mutated EPHA2. Our findings demonstrated that EPHA2 mutants may be an attractive therapeutic target for lymphatic metastasis of ICC.

Osteopontin promotes hepatocellular carcinoma progression via the PI3K/AKT/Twist signaling pathway.

The epithelial-mesenchymal transition (EMT) serves critical roles in the migration, invasion and metastasis of human cancer cells. This process is initiated by regulation of E-cadherin expression by the major inducers of EMT. Previous studies reported that osteopontin (OPN) is essential for hepatocellular carcinoma (HCC) metastasis as it facilitates the EMT in HCC. However, the role and clinical significance of OPN as an EMT regulator in HCC remains unknown. The present study revealed that OPN regulated the expression of Twist by activating RAC serine/threonine-protein kinase (Akt), a critical EMT regulator. Interfering with the phosphoinositide 3-kinase (PI3K)/Akt pathway may suppress the expression of Twist enhanced by OPN. Increased Twist levels in HCC were associated with poor survival and tumor recurrence in patients with HCC following surgery. A significant association was observed between OPN expression and Twist levels in HCC, and a combination of these two parameters was revealed to be a more powerful predictor of poor patient prognosis. The findings of the present study indicate that Twist serves an notable role in OPN-mediated metastasis of HCC through activation of the PI3K/Akt pathway. Twist may be a potential therapeutic target for the prevention of HCC metastasis in patients exhibiting high OPN expression.

Osteopontin alters DNA methylation through up-regulating DNMT1 and sensitizes CD133+/CD44+ cancer stem cells to 5 azacytidine in hepatocellular carcinoma.

BACKGROUND: In hepatocellular carcinoma (HCC), CD133+/CD44+ cells are one subgroup with high stemness and responsible for metastatic relapse and resistance to treatment. Our previous studies have demonstrated that osteopontin (OPN) plays critical roles in HCC metastasis. We further investigated the molecular mechanism underlying the role of OPN in regulating the stemness of HCC epigenetically and explored possible targeting strategy. METHODS: CD133+/CD44+ subgroup sorting from HCC cell lines and HCC tissues was used to investigate the effects of OPN knockdown on stemness. iTRAQ and MedIP-sequencing were applied to detect the protein profile and epigenetic modification of CD133+/CD44+ subgroup with or without OPN knockdown. The antitumor effects of 5 Azacytidine were examined in cultured HCC cells and patient derived xenograft (PDX) models. RESULTS: OPN was accumulated in CD133+/CD44+ subgroup of HCC cells. Knocking down OPN significantly inhibited the sphere formation and stemness-related genes expression, and delayed tumor initiation of CD133+/CD44+ subgroup of HCC cells. Employing MedIP-sequencing, dot blot and iTRAQ analyses of CD133+/CD44+ SCR and CD133+/CD44+ shOPN cells, we found that OPN knockdown leaded to reduction in DNA methylation with particular enrichment in CGI. Meanwhile, DNA (cytosine-5)-methyltransferase 1 (DNMT1), the main methylation maintainer, was downregulated via proteomics analysis, which mediated OPN altering DNA methylation. Furthermore, DNMT1 upregulation could partially rescue the properties of CD133+/CD44+ shOPN cells. Both in vitro and in vivo assays showed that CD133+/CD44+ cells with high OPN levels were more sensitive to DNA methylation inhibitor, 5 Azacytidine (5 Aza). The above findings were validated in HCC primary cells, a more clinically relevant model. CONCLUSIONS: OPN induces methylome reprogramming to enhance the stemness of CD133+/CD44+ subgroup and provides the therapeutic benefits to DNMT1 targeting treatment in HCC.

MicroRNA-219-5p Promotes Tumor Growth and Metastasis of Hepatocellular Carcinoma by Regulating Cadherin 1.

MicroRNAs play significant roles in the development of cancer and may serve as promising therapeutic targets. In our previous work, miR-219-5p was identified as one of the important metastasis-related microRNAs in HCC. Here we demonstrated that miR-219-5p expression was elevated in HCC tissues and was associated with vascular invasion and dismal prognosis. In multivariate analysis, miR-219-5p was identified as an independent prognostic indicator for HCC patients. Functional mechanism analyses showed that miR-219-5p promoted HCC cell proliferation and invasion in in vitro, as well as in vivo, tumor growth and metastasis in nude mice models bearing human HCC tumors. In addition, cadherin 1 (CDH1) was revealed to be a downstream target of miR-219-5p in HCC cells. In conclusion, miR-219-5p promotes tumor growth and metastasis of HCC by regulating CDH1 and can serve as a prognostic marker for HCC patients.

The dual blockade of MET and VEGFR2 signaling demonstrates pronounced inhibition on tumor growth and metastasis of hepatocellular carcinoma.

BACKGROUND: The application of VEGF signaling inhibitors have been associated with more invasive or metastatic behavior of cancers including hepatocellular carcinoma (HCC). We explored the contribution of MET pathway to the enhanced HCC invasion and metastasis by VEGF signaling inhibition, and investigated the antitumor effects of NZ001, a novel dual inhibitor of MET and VEGFR2, in HCC. METHODS: Immunocompetent orthotopic mice model of hepal-6 was established to investigate the effects of either VEGF antibody alone or in combination with the selective MET inhibitor on tumor aggressiveness. The antitumor effects of NZ001 were examined in cultured HCC cells as well as in vivo models. MET gene amplification was determined by SNP 6.0 assay. MET/P-MET expression was detected by IHC. RESULTS: Selective VEGF signaling inhibition by VEGF antibody significantly reduced in vivo tumor growth of the orthotopic mice models, simultaneously also enhanced tumor invasion and metastasis, but inhibiting MET signaling attenuated this side-effect. Further study revealed that hypoxia caused by VEGF signaling inhibition induced HIF-1α nuclear accumulation, subsequently leading to elevated total-MET expression, and synergized with HGF in inducing invasion. NZ001, a novel dual inhibitor of MET and VEGFR2, markedly inhibited both tumor growth and metastasis of HCC, which showed obvious advantages over sorafenib in not inducing more invasive and metastatic behaviors. This effect is more pronounced in HCC with MET amplification and overexpression. CONCLUSIONS: The activation of MET is responsible for the metastasis-promoting effects induced by VEGF inhibition. MET and VEGFR2 dual blockade, NZ001, has advantages over sorafenib in not inducing more invasive and metastatic behaviors; MET amplification and overexpression can be used to identify the subgroup of patients most likely to get the optimal benefit from NZ001 treatment.

Osteopontin promotes metastasis of intrahepatic cholangiocarcinoma through recruiting MAPK1 and mediating Ser675 phosphorylation of ß-Catenin.

The incidence and mortality of intrahepatic cholangiocarcinoma (ICC) are increasing worldwide in recent decades. Osteopontin (OPN) plays an important role in cancer metastasis, but its functional mechanism in ICC is not clear yet. In this study, we found that OPN level was elevated both in plasma and tumor tissues of ICC patients, which was closely related to a shorter overall survival (OS) and high probability of tumor relapse after curative resection. The gain- and loss-of-function studies determined that OPN could promote ICC growth and metastasis. OPN selectively interacted with ß-Catenin and knockdown of ß-Catenin abrogated the effects induced by OPN. OPN recruited MAPK1 and activated MEK-MAPK1 pathway to mediate the S675 phosphorylation of ß-Catenin and nucleus accumulation, which induced the activation of Wnt signaling. Moreover, a significant correlation between OPN and ß-Catenin was found in ICC tissues. OPN, ß-Catenin, and their combination were independent prognostic indicator for ICC patients. In conclusion, OPN promotes ICC progression through recruiting MAPK1 and activating the Wnt/ß-Catenin pathway and can serve as a novel prognostic marker and therapeutic target for ICC.

Whole-exome mutational and transcriptional landscapes of combined hepatocellular cholangiocarcinoma and intrahepatic cholangiocarcinoma reveal molecular diversity.

BACKGROUND: Primary liver cancer (PLC) is the third largest contributor to cancer mortality in the world. PLC is a heterogeneous disease that encompasses several biologically distinct subtypes including hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (ICC) and combined hepatocellular-cholangiocarcinoma (CHC). CHC is a distinct, albeit rare, subtype of PLC and is comprised of cells with histopathological features of both HCC and ICC. Several studies have focused on the mutation and expression landscapes of HCC and ICC. However, studies of CHC were rare. OBJECTIVE: The aim of the current study was to identify genetic and gene expression alterations in the carcinogenesis and development of CHC and ICC in the Chinese population. Unraveling both similar and differing patterns among these subtypes may help to identify personalized medicine approaches that could improve patient survival. METHODS: Whole genome sequencing (WGS), whole exome sequencing (WES) and RNA-seq were performed on 10 ICC and 10 CHC samples, matched with adjacent non-tumor liver tissue specimens. Comparative analysis was performed using HCC datasets from The Cancer Genome Atlas (TCGA). RESULTS: Mutational and transcriptional landscapes of CHC and ICC were clearly delineated. TP53 and CTNNB1 were identified as exhibiting mutations in CHC. ARID1A, PBRM1, and IDH1 were frequently mutated in ICC. RYR3, FBN2, and KCNN3 are associated with cell migration and metastasis and might be driver genes in CHC. KCNN3 was identified as also exhibiting mutations in ICC. The ECM-receptor interaction pathway associated fibrogenic hepatic progenitor cell differentiation and liver fibrosis may play an important role in carcinogenesis of PLC. Chromatin remodeling and chromosome organization are key processes in carcinogenesis and development in PLC. P53 related pathways showed alterations in CHC and HCC. Inflammation may be a key factor involved in ICC carcinogenesis. CONCLUSION: CHC and ICC are different subtypes of PLC. This study discusses predominantly the molecular genetic details of PLC subtypes and highlights the need for an accurate diagnosis and treatment of specific PLC subtypes to optimize patient management.