DCE-MRI-based radiomics in predicting angiopoietin-2 expression in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is the sixth most common cancer, and the third leading cause of cancer death worldwide. Studies have shown that increased angiopoietin-2 (Ang-2) expression relative to Ang-1 expression in tumors is associated with a poor prognosis.The purpose of this study was to investigate the efficacy of predicting Ang-2 expression in HCC by preoperative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI)-based radiomics. METHODS: The data of 52 patients with HCC who underwent surgical resection in our hospital were retrospectively analyzed. Ang-2 expression in HCC was analyzed by immunohistochemistry. All patients underwent preoperative upper abdominal DCE-MRI and intravoxel incoherent motion diffusion-weighted imaging scans. Radiomics features were extracted from the early and late arterial and portal phases of axial DCE-MRI. Univariate analysis and least absolute shrinkage and selection operator (LASSO) was performed to select the optimal radiomics features for analysis. A logistic regression analysis was performed to establish a DCE-MRI radiomics model, clinic-radiologic (CR) model and combined model integrating the radiomics score with CR factors. The stability of each model was verified by 10-fold cross-validation. Receiver operating characteristic (ROC) curve analysis, calibration curve analysis and decision curve analysis (DCA) were employed to evaluate these models. RESULTS: Among the 52 HCC patients, high Ang-2 expression was found in 30, and low Ang-2 expression was found in 22. The areas under the ROC curve (AUCs) for the radiomics model, CR model and combined model for predicting Ang-2 expression were 0.800, 0.874, and 0.933, respectively. The DeLong test showed that there was no significant difference in the AUC between the radiomics model and the CR model (p > 0.05) but that the AUC for the combined model was significantly greater than those for the other 2 models (p < 0.05). The DCA results showed that the combined model outperformed the other 2 models and had the highest net benefit. CONCLUSION: The DCE-MRI-based radiomics model has the potential to predict Ang-2 expression in HCC patients; the combined model integrating the radiomics score with CR factors can further improve the prediction performance.

Radiomics models based on multisequence MRI for predicting PD-1/PD-L1 expression in hepatocellular carcinoma.

The purpose of this study was to explore the effectiveness of radiomics based on multisequence MRI in predicting the expression of PD-1/PD-L1 in hepatocellular carcinoma (HCC). One hundred and eight patients with HCC who underwent contrast-enhanced MRI 2 weeks before surgical resection were enrolled in this retrospective study. Corresponding paraffin sections were collected for immunohistochemistry to detect the expression of PD-1 and PD-L1. All patients were randomly divided into a training cohort and a validation cohort at a ratio of 7:3. Univariate and multivariate analyses were used to select potential clinical characteristics related to PD-1 and PD-L1 expression. Radiomics features were extracted from the axial fat-suppression T2-weighted imaging (FS-T2WI) images and the arterial phase and portal venous phase images from the axial dynamic contrast-enhanced MRI, and the corresponding feature sets were generated. The least absolute shrinkage and selection operator (LASSO) was used to select the optimal radiomics features for analysis. Logistic regression analysis was performed to construct single-sequence and multisequence radiomics and radiomic-clinical models. The predictive performance was judged by the area under the receiver operating characteristic curve (AUC) in the training and validation cohorts. In the whole cohort, PD-1 expression was positive in 43 patients, and PD-L1 expression was positive in 34 patients. The presence of satellite nodules served as an independent predictor of PD-L1 expression. The AUC values of the FS-T2WI, arterial phase, portal venous phase and multisequence models in predicting the expression of PD-1 were 0.696, 0.843, 0.863, and 0.946 in the training group and 0.669, 0.792, 0.800 and 0.815 in the validation group, respectively. The AUC values of the FS-T2WI, arterial phase, portal venous phase, multisequence and radiomic-clinical models in predicting PD-L1 expression were 0.731, 0.800, 0.800, 0.831 and 0.898 in the training group and 0.621, 0.743, 0.771, 0.810 and 0.779 in the validation group, respectively. The combined models showed better predictive performance. The results of this study suggest that a radiomics model based on multisequence MRI has the potential to predict the preoperative expression of PD-1 and PD-L1 in HCC, which could become an imaging biomarker for immune checkpoint inhibitor (ICI)-based treatment.

A correlative study between IVIM-DWI parameters and VEGF and MMPs expression in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer-related death worldwide. Angiogenic factors may be valuable indices of tumor recurrence and treatment and potentially useful markers for predicting the response to antiangiogenesis therapy. Vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs) are major drivers of tumor angiogenesis. Preoperatively predicting the expression of VEGF and MMPs is crucial for treating HCC. Intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) has been successfully used in the differential diagnosis of HCC, pathological grading, and treatment response evaluation. However, the correlations between IVIM-DWI parameters and VEGF and MMP expression have not been reported. This study provides a preliminary analysis of the correlation between IVIM-DWI parameters and the expression of VEGF, MMP-2, and MMP-9 to investigate the value of IVIM-DWI in the noninvasive evaluation of angiogenesis in HCC. Methods: IVIM-DWI was performed in 61 patients with HCC 1 week before they underwent surgical resection. VEGF, MMP-2, and MMP-9 expression was detected using immunohistochemistry staining. Spearman correlation analysis was used to analyze the correlations between the IVIM-DWI parameters and VEGF, MMP-2, and MMP-9 expression in HCC. Results: The fast apparent diffusion coefficient fraction (f) value was positively correlated with the expression of VEGF (P<0.001), MMP-2 (P=0.002), and MMP-9 (P<0.001). The fast apparent diffusion coefficient (D*) was positively correlated with VEGF (P<0.001) and MMP-9 (P<0.001) expression but was not correlated with MMP-2 (P=0.659) expression. The apparent diffusion coefficient (ADC) and slow apparent diffusion coefficient (D) values were not significantly correlated with the expression of VEGF (P=0.103 and P=0.543, respectively), MMP-2 (P=0.596 and P=0.338, respectively), or MMP-9 (P=0.102 and P=0.660, respectively). Conclusions: IVIM-DWI can be used to noninvasively evaluate angiogenesis in HCC.

Radiomic Analysis Based on Magnetic Resonance Imaging for Predicting PD-L2 Expression in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is the sixth most common malignant tumour and the third leading cause of cancer death in the world. The emerging field of radiomics involves extracting many clinical image features that cannot be recognized by the human eye to provide information for precise treatment decision making. Radiomics has shown its importance in HCC identification, histological grading, microvascular invasion (MVI) status, treatment response, and prognosis, but there is no report on the preoperative prediction of programmed death ligand-2 (PD-L2) expression in HCC. The purpose of this study was to investigate the value of MRI radiomic features for the non-invasive prediction of immunotherapy target PD-L2 expression in hepatocellular carcinoma (HCC). A total of 108 patients with HCC confirmed by pathology were retrospectively analysed. Immunohistochemical analysis was used to evaluate the expression level of PD-L2. 3D-Slicer software was used to manually delineate volumes of interest (VOIs) and extract radiomic features on preoperative T2-weighted, arterial-phase, and portal venous-phase MR images. Least absolute shrinkage and selection operator (LASSO) was performed to find the best radiomic features. Multivariable logistic regression models were constructed and validated using fivefold cross-validation. The area under the receiver characteristic curve (AUC) was used to evaluate the predictive performance of each model. The results show that among the 108 cases of HCC, 50 cases had high PD-L2 expression, and 58 cases had low PD-L2 expression. Radiomic features correlated with PD-L2 expression. The T2-weighted, arterial-phase, and portal venous-phase and combined MRI radiomics models showed AUCs of 0.789 (95% CI: 0.702-0.875), 0.727 (95% CI: 0.632-0.823), 0.770 (95% CI: 0.682-0.875), and 0.871 (95% CI: 0.803-0.939), respectively. The combined model showed the best performance. The results of this study suggest that prediction based on the radiomic characteristics of MRI could noninvasively predict the expression of PD-L2 in HCC before surgery and provide a reference for the selection of immune checkpoint blockade therapy.

Overexpression of cytoplasmic sphingosine 1-phosphate receptor 1 promotes cell cycle progression and migration in human esophageal squamous cell carcinoma.

Sphingosine-1-phosphate receptor 1 (S1PR1) is abnormally expressed in a variety of tumors. However, the clinical implications and biological roles of S1PR1 in esophageal squamous cell carcinoma (ESCC) remain unknown. In this study, we have focused on ESCC, and analyzed the expression of S1PR1 in human specimens at various histological grades of ESCC and the role of S1PR1 in Eca109 cells. Using human ESCC tissue microarray and immunohistochemistry, we found S1PR1 protein mainly located in the cytoplasm of cancer cells and normal esophageal mucosal epithelial cells, and small amounts in the plasma membrane. The levels of cytoplasmic S1PR1 in ESCC tissues were significantly higher than those in adjacent non-cancerous tissues. Cytoplasmic S1PR1 exhibited higher expression in ESCC tissues with poor differentiation than those with well differentiation. Conversely, the positive expression of plasma membrane S1PR1 was correlated with well differentiation. Kaplan-Meier survival analysis showed that patients with positive membrane S1PR1 expression tended to have longer survival time. Univariate and multivariate Cox regression analysis revealed that membrane S1PR1 expression was an independent prognostic factor for ESCC patients. Furthermore, overexpression of cytoplasmic S1PR1 promoted Eca109 cells from G1 phase to S phase and plasma membrane S1PR1 as the opposite, which may be associated with p21. Cytoplasmic S1PR1 signaling also promoted Eca109 cells migration. Our findings demonstrate that cytoplasmic S1PR1 plays an important role in the malignant behavior of human ESCC and may serve as a new target for ESCC therapy.

Human infection with a novel, highly pathogenic avian influenza A (H5N6) virus: Virological and clinical findings.

BACKGROUND AND OBJECTIVES: Severe infection with avian influenza A (H5N6) virus in humans was identified first in 2014 in China. Before that, it was unknown or unclear if the disease or the pathogen affected people. This study illustrates the virological and clinical findings of a fatal H5N6 virus infection in a human patient. METHODS: We obtained and analyzed the clinical, epidemiological, and virological data from the patient. Reverse transcription polymerase chain reaction (RT-PCR), viral culture, and sequencing were conducted for determination of the causative pathogen. RESULTS: The patient, who presented with fever, severe pneumonia, leucopenia, and lymphopenia, developed septic shock and acute respiratory distress syndrome (ARDS), and died on day 10 after illness onset. A novel reassortant avian-origin influenza A (H5N6) virus was isolated from the throat swab or trachea aspirate of the patient. The virus was reassorted with the HA gene of clade 2.3.4.4 H5, the internal genes of clade 2.3.2.1 H5, and the NA gene of the H6N6 avian virus. The cleavage site of the HA gene contained multiple basic amino acids, indicating that the novel H5N6 virus was highly pathogenic in chicken. CONCLUSIONS: A novel, highly pathogenic avian influenza H5N6 virus with a backbone of H5N1 virus acquired from the NA gene from the H6N6 virus has been identified. It caused human infection resulting in severe respiratory disease.

[The apoptosis effect of antisense oligodeoxynucleotides targeting ATM on the laryngeal squamous cell carcinoma treated with radiation].

OBJECTIVE: To designed antisense oligodeoxynucleotides (AS-ODNs) to reduce the expression of ATM and to study its effect on the apoptosis of Hep-2 (human epidermoid laryngeal carcinoma) cells treated with radiation in vitro. METHODS: Experiment was divided into AS-Lipo, Sen-Lipo, Mis-Lipo, Lipo and Hep-2 group. The expression of ATM mRNA in Hep-2 cells was examined by real-time quantitative PCR. About 18 hours after transfection, they were irradiated simultaneously with different doses of X-ray radiation (0, 2, 4, 6, and 8 Gy) respectively. Clonogenic survival assay was carried out to detect the survival ability of Hep-2 cells after irradiation. After exposed to 4 Gy radiation, flow cytometry was carried out to analyze the cell apoptosis. RESULTS: The relative ATM mRNA expression in Hep-2 cells treated with ATM AS-ODNs was decreased to (11.03 +/- 2.51)% which was much lower than that of untreated cells (P < 0.05). After irradiation, the survival fraction (SF) of cells treated with ATM AS-ODNs was lower than that of other groups at the same dose of radiation. There was statistical significance between the group treated with ATM AS-ODNs and other groups (P < 0.05). The apoptotic rate for the group irradiated with ATM AS-ODNs was (30.7 +/- 1.31)%, which was significantly higher than that of others (P < 0.05). CONCLUSION: AS-ODNs of ATM reduce ATM mRNA expression and enhance Hep-2 cells apoptosis to radiation in vitro.

[Enhanced expression of human vimentin intermediate filaments in hepatocellular carcinoma cells decreases their proliferative and invasive abilities in vitro].

OBJECTIVE: Expression of vimentin in carcinoma cells is a marker for poor prognosis in patients. The aim of this investigation was to assess the influence of vimentin on the characteristics of carcinoma cells. METHODS: The full-length vimentin gene open reading frame (1401 base pairs) was cloned into the plasmid vector pcDNA 3.1 (+), and these vectors were used to stably transfect the human hepatocellular carcinoma HepG2 cell line. Vimentin gene expression was evaluated by RT-PCR and Western blot. Proliferative activity and invasive potential of tumor cells were determined by the CellTiter 96 aqueous one solution cell proliferation assay and BioCoat GFR Matrigel invasion chamber, respectively. RESULTS: DNA sequencing and restriction endonuclease digestion analysis demonstrated that the recombinant vector was correctly cloned. The stable cell line demonstrated a higher vimentin RNA and protein expression. However, both proliferative and invasive abilities of the cells were reduced in vitro ( P < 0.05). CONCLUSION: A recombinant plasmid pcDNA3. 1-VIM is successfully constructed and a carcinoma cell line HepG2-pV highly expressing vimentin is obtained. Recombinant vimentin protein suppresses the proliferative and invasive abilities of HepG2 cells, suggesting that it might decrease malignant phenotype of tumor cells in vitro. This work makes a foundation for further study on the relationship between vimentin and biological phenotype of carcinoma cells.

[Effect of inhibitor HNP1 transfection on tumor growth of human nasopharyngeal carcinoma cell line HNE-1].

OBJECTIVE: The aim of this study is to investigate the effects of inhibitor HNP1 transfection on proliferation and tumor growth of human nasopharyngeal carcinoma cell line HNE1. METHOD: HNE1 cells were transfected with HNP1 by liposome, and the cytotoxic effect was detected by MTT, In vivo tumor growth test was performed in nude mice inoculated with transfected HNE1 cells. The therapeutic effect of HNP1 was evaluated in the inoculated tumors, alpha-defensin 1 expression was detected in implanted tumor tissues by immunohistochemical stain. RESULT: HNP1 transfection significantly inhibited the proliferation of HNE1 cells. MTT assay confirmed cytotoxic effect. In vivo study showed tumor volume was significantly smaller in HNP1 transfection group than that in control group (P < 0.01). Immunohistochemical stain showed alpha-defensin expression was increased in HNP1 transfection group. CONCLUSION: HNP1 transfection can inhibit the proliferation of HNE1 cells, as well as tumor growth.

[Searching for genes interacting with human PCIA1 gene by using the bacterial two-hybrid system].

OBJECTIVE: To search for the genes which could interact with newly found homo sapiens cross-immune reaction antigen (PCIA1) gene and accordingly to provide insights into the study of the gene function. METHODS: The Stratagene's BacterioMatch Two-Hybrid System and BacterioMatch Fetal Kidney Library were adopted and the recombinant bait plasmid pBT-PCIA1 was cotransformated with the target plasmid pTRG-cDNA library DNA into the reporter stain. After screening and isolation of positive pTRG clones, the target genes were identified by DNA sequencing and bioinformation analysis. RESULTS: Among all the seven detected target genes, three genes' function were not known, the other four genes had important functions. Their mutations or abberant expression resulted in severe diseases and overexpression of ACTN4 (actinin, alpha 4), PSAP (prosaposin) or EIF3S10 (eukaryotic translation initiation factor 3, subunit 10 theta) could promote tumor development and progression. CONCLUSION: The bacterial two-hybrid system technique is an efficient method, which can provides insights into the study of novel genes' function by detecting protein-protein interactions. This study indicates that PCIA1 gene expression correlates with tumor formation, invasion and metastasis.

Up-regulation of gastric cancer cell invasion by Twist is accompanied by N-cadherin and fibronectin expression.

Twist, a newly found EMT-inducer, has been reported to be up-regulated in those of diffuse-type gastric carcinomas with high N-cadherin level. We show here MKN45, a cell line derived from undifferentiated carcinomas cells, expresses high levels of Twist. Down-regulation of Twist, using an antisense Twist vector in MKN45 cells, inhibits cell migration and invasion, companied with a morphologic changes associated with MET. Suppression of Twist also decreases the expressions of N-cadherin and fibronectin, but not of E-cadherin in MKN45. In contrast, overexpression of Twist in MKN28, a cell line derived from moderate differentiated carcinomas, results in up-regulation of N-cadherin and fibronectin, companied with down-regulation of E-cadherin. Taken together, our results suggest that Twist regulates cell motility and invasion in gastric cancer cell lines, probably through the N-cadherin and fibronectin production.

[Expression of heat shock protein-70, estrogen receptor and progesterone receptor in ovarian carcinomas and the correlation between HSP70 and sex steroid receptor].

OBJECTIVE: This study was aimed to determine the expression of HSP70, ER and PR in ovarian carcinomas and to explore the relationship between HSP70 and sex steroid receptor. METHODS: The immunohistochemical way SP was performed to estimate the expression of HSP70, ER and PR in 41 cases of ovarian carcinomas and in 11 cases of normal ovarian tissue. RESULTS: The positive staining rate of HSP70 was 68.29% (28/41), which was remarkably higher than that in normal ovarian tissue (18.18%) (P<0.05). Furthermore, the expression rate of HSP70 was much higher in poorly differentiated ovarian carcinomas than in well differentiated ovarian carcinomas (P<0.05). ER positive staining was observed in 19 cases (46.34%), and PR in 24 cases (58.54%). ER and PR positive staining occurred more frequently in the group of HSP70 negative staining than in the group of HSP70 positive staining. related with the expression of PR (P<0.05). CONCLUSION: The expression of HSP70 was negatively related with the expression of PR (P<0.05).