[Circular RNA-Encoded Proteins in Gastrointestinal Cancer:A Review].

Circular RNAs(CircRNAs)are a class of non-coding RNAs with a covalently closed-loop structure,high stability,and tissue specificity,with the production mechanisms different from linear RNAs.Recent studies have discovered that some CircRNAs can encode proteins via cap-independent translation mechanisms such as internal ribosome entry site,N6-methyladenosine,and rolling loop translation.The encoded proteins regulate homologous linear proteins or downstream signaling pathways via protein bait or other mechanisms,thereby exerting biological functions.Studies have shown that CircRNAs play a role in various diseases,especially in tumor progression,proliferation,invasion,and metastasis and immune regulation.Therefore,by elucidating the expression and roles of proteins encoded by CircRNAs in tumorigenesis and development,this paper is expected to provide new tumor markers and potential targets for tumor diagnosis and treatment.

Immunomodulatory effect of Myrtenol on benzo (a) pyrene-induced lung cancer in Swiss albino mice via modulation of tumor markers, cytokines and inhibition of PCNA expression.

Lung cancer is one of the most common cancers in men. Although many diagnostic and treatment regimens have been followed in the treatment for lung cancer, increasing mortality rate due to lung cancer is depressing and hence requires alternative plant based therapeutics with with less side-effects. Myrtenol exhibits anti-inflammatory and antioxidant properties. Hence we intended to study the effect of Myrtenol on B(a)P-induced lung cancer. Our study showed that B(a)P lowered hematological count, decreased phagocyte and avidity indices, nitroblue tetrazolium (NBT) reduction, levels of immunoglubulins, antioxidant levels, whereas Myrtenol treatment restored them back to normal levels. On the other hand, xenobiotic and liver dysfunction marker enzymes and pro-inflammatory cytokines were elevated on B(a)P exposure, which retuned back to normal by Myrtenol. This study thus describes the immunomodulatory and antioxidant effects of Myrtenol on B[a]P-induced immune destruction.

A bibliometric study and visualization analysis of ferroptosis-inducing cancer therapy.

Ferroptosis is a form of regulated cell death that was first formally proposed a decade ago. While its role in cancer cell death was initially understudied, it has recently gained considerable interest from researchers. In recent years, a growing number of studies have focused on the role of ferroptosis in cancer progression, with the goal of developing novel ferroptosis-inducing cancer therapies. This study aims to present the developmental trend and hotspots of research on ferroptosis-inducing cancer therapy using bibliometric analysis. A literature search was conducted using the Web of Science Core Collection on October 1st, 2022, to retrieve articles and reviews pertaining to ferroptosis and cancer published from 2012 to 2022. Microsoft Excel 2016, VOSviewer 1.6.18 and CiteSpace (version 6.1. R6) were utilized to conduct the bibliometric analysis of publication trends, authorship, and citation networks, with a focus on identifying countries, institutions, journals, and authors contributing to the field. These analyses were used to predict future trends in this area. A total of 2839 articles were identified and extracted for analysis. The number of publications has increased almost every year, with a sharp increase after 2018. China produced the most publications in this area, followed by the United States. Central South University was the institution that published the most papers. Frontiers in Oncology was the journal with the highest number of publications, while Cell had the greatest impact factor. Daolin Tang was the most productive author and Dixon SJ was the most influential author. Co-occurrence and burst analyses of keywords and references were conducted to identify the developmental trends and hotspots in ferroptosis-inducing cancer therapy research. Main research directions have shifted from investigating the mechanism of ferroptosis to developing novel ferroptosis-targeting cancer therapies. Emerging topicsfocus on the role of ferroptosis in solid tumor therapy. Based on our bibliometric analysis, we predict that research on ferroptosis in cancer therapy will continue to be a hot topic in the future, with a growing number of treatment modalities related to ferroptosis being developed. Our study provides valuable insights into the current state and future trends of research in this field, serving as a useful guide for researchers seeking to make important contributions in this area.

Knockdown of long non-coding RNA LINC00941 suppressed cell proliferation, colony-formation, and migration of human glioblastoma cell lines.

INTRODUCTION: Glioblastoma (GBM) represents the most common and lethal type of primary brain tumour in adults, and due to its high invasiveness, treatment of GBM remains challenging. This work is aimed to elucidate the role of LINC00941 in GBM. MATERIAL AND METHODS: Expression of LINC00941 in two GBM cell lines U251 and U87-MG was knocked down using siRNA. Cell proliferation and colony-formation ability of LINC00941 knockdown were examined. Apoptosis of the knockdown was evaluated using flow cytometry, with the levels of Bax, Bcl-2, cleaved caspase-3, and phosphorylation of ERK and Akt to be examined using western blotting. Migration and invasion of the knockdown was studied using transwell assays. RESULTS: Expression of LINC00941 was significantly elevated in GBM compared to non-tumour tissues ( p < 0.01). Statistical analysis on the expression data further revealed the negative correlation between LINC00941 and miR-526b-5p ( r = 0.7494, p < 0.001). LINC00941 was successfully knocked down with RNA interference in U251 and U87-MG. The knockdown significantly suppressed cell proliferation and the ability to form colonies. Percentage of apoptotic cells was elevated by the knockdown in both cell lines as evidenced by flow cytometric analysis, which was accompanied by a significant decrease in Bcl-2 and substantial increases in Bax and cleaved caspase-3. Phosphorylation of ERK and Akt was also enhanced in both cell lines by the knockdown. In addition, knockdown of LINC00941 suppressed migration of both cell lines across transwell membrane and matrigel. CONCLUSIONS: LINC00941 is overexpressed in GBM, exhibiting important roles in cell proliferation and survival, migration and invasion.

Research on rainy day traffic sign recognition algorithm based on PMRNet.

The recognition of traffic signs is of great significance to intelligent driving and traffic systems. Most current traffic sign recognition algorithms do not consider the impact of rainy weather. The rain marks will obscure the recognition target in the image, which will lead to the performance degradation of the algorithm, a problem that has yet to be solved. In order to improve the accuracy of traffic sign recognition in rainy weather, we propose a rainy traffic sign recognition algorithm. The algorithm in this paper includes two modules. First, we propose an image deraining algorithm based on the Progressive multi-scale residual network (PMRNet), which uses a multi-scale residual structure to extract features of different scales, so as to improve the utilization rate of the algorithm for information, combined with the Convolutional long-short term memory (ConvLSTM) network to enhance the algorithm's ability to extract rain mark features. Second, we use the CoT-YOLOv5 algorithm to recognize traffic signs on the recovered images. In this paper, in order to improve the performance of YOLOv5 (You-Only-Look-Once, YOLO), the 3 × 3 convolution in the feature extraction module is replaced by the Contextual Transformer (CoT) module to make up for the lack of global modeling capability of Convolutional Neural Network (CNN), thus improving the recognition accuracy. The experimental results show that the deraining algorithm based on PMRNet can effectively remove rain marks, and the evaluation indicators Peak Signal-to-Noise Ratio (PSNR) and Structural Similarity Index Measure (SSIM) are better than the other representative algorithms. The mean Average Precision (mAP) of the CoT-YOLOv5 algorithm on the TT100k datasets reaches 92.1%, which is 5% higher than the original YOLOv5.

Retrospective Analysis of Postoperative Effect of Supratubal Recess Opened and Bony Obliteration Tympanoplasty.

BACKGROUND: In the surgical development of cholesteatoma, in order to reduce the recurrence of cholesteatoma, 2 kinds of surgeries were carried out: removal of Cog and Korner's septum to ventilate supratubal recess (supratubal recess opened) and obliteration of the mastoid and attic space (bony obliteration tympanoplasty) were invented, respectively. Their purpose is the same but the theoretical basis is different, and the comparison of these 2 methods is not reported in the current literature. This study aims to evaluate the rates of recurrent and residual cholesteatoma with the simple canal wall up and canal wall up-supratubal recess opened and canal wall up-bony obliteration tympanoplasty mastoidectomy in a large cohort of patients. The secondary objectives were to assess the 3 techniques' infection rates and hearing outcomes. METHODS: Overall, 352 patients with middle ear cholesteatoma preoperatively underwent temporal bone ultrahigh-resolution computed tomography scan. The shape of the Eustachian tube and the supratubal recess were analyzed, and superior and posterior tympanic recesses, including the supratubal recess, were opened in different surgical groups. RESULTS: After 5 years of follow-up, the results show that the lowest recurrence rate was 6.6% (7/106) for canal wall up-supratubal recess opened, 10.9% (12/101) for canal wall up-bony obliteration tympanoplasty, and canal wall up had the highest recurrence rate of 19.31% (28/145). The postoperative infection rate was 5.7% in the canal wall up-supratubal recess opened group, 10.89% in the canal wall up-bony obliteration tympanoplasty group, and 7.59% in the simple canal wall up group. The postoperative median air conduction was increased 8 dB in the canal wall up-supratubal recess opened group, 1 dB in the canal wall up-bony obliteration tympanoplasty, and 6 dB in the simple canal wall up group. CONCLUSION: Opening the supratubal recess to ensure the patency of the attic facilitates the gas exchange between the mastoid process and the middle ear and reduces the possibility of cholesteatoma recurrence.

Long non-coding RNA-LINC00941 promotes the proliferation and invasiveness of glioma cells.

Glioma is one of the most common intracranial malignant tumors worldwide, accounting for 30%-40% of primary brain tumors. Long non-coding RNAs (lncRNAs) have been implicated in cancer malignant progression. Glioma is classified into multiple subtypes, but lncRNA expression pattern in different subtypes are not fully described. Here, we reported that lncRNA-LINC00941 was highly expressed in all glioma subtypes. Overexpression of lncRNA-LINC00941 in U87 cells promoted cellular proliferation and invasiveness, and suppressed apoptosis. Our findings suggest that lncRNA-LINC00941 may function as an oncogenic factor in glioma, and targeting lncRNA-LINC00941 could be developed into a strategy for glioma management.

ADAM10-cleaved ephrin-A5 contributes to prostate cancer metastasis.

A disintegrin and metalloprotease-10(ADAM10) promotes the metastasis of prostate cancer (PCa), but the specific mechanism is indistinct. Herein, DU145 cell lines with stable overexpression and knockdown of ADAM10 were constructed. We found that ectopic expression of ADAM10 not only significantly facilitated cell proliferation, migration, invasion, and inhibited apoptosis, but also could specifically hydrolyze ephrin-A5 and release the ephrin-A5 soluble ectodomain into extracellular media in vitro. These effects were reversed by ADAM10 depletion or treatment of GI254023X. Meanwhile, the co-location and physical interaction among EphA3, ephrin-A5, and ADAM10 were observed in PCa cells using immunofluorescence and immunoprecipitation techniques. Interestingly, overexpression of EphA3 exerted opposite effects in DU145 (ephrin-A5 + ) cells and PC-3 (ephrin-A5 ± ) cells. In addition, the pro-tumor function of EphA3 was reversed by the treatment with the exogenous ephrin-A5-Fc, which increased the phosphorylation level of EphA3 in PC-3 (ephrin-A5 ± ) cells. In nude mice, ADAM10 accelerated growth of the primary tumor, decreased the level of ephrin-A5 in the tumor tissue, but increased the level of ephrin-A5 in the peripheral blood, accompanied with an increase in the expression of CD31 and VEGF (vascular endothelial growth factor) in the tissue. What is more, the serum ephrin-A5 content of patients with metastatic PCa was significantly higher than that of the non-metastatic group (P < 0.05). The receiver operating characteristic curve(ROC) showed that the area under the curve(AUC) of serum ephrin-A5 as a marker of PCa metastasis was 0.843, with a sensitivity of 93.5% and a specificity of 75%. It is concluded that ADAM10-mediated ephrin-A5 shedding promotes PCa metastasis via transforming the role of EphA3 from ligand-dependent tumor suppressor to ligand-independent promoter, and ephrin-A5 in the blood can be used as a new biomarker for PCa metastasis.

Bioinformatics Methods Reveal the Biomarkers and the miRNA-mRNA Network in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) has threatened the health of humans, and few therapeutic strategies can completely uproot this illness. Bioinformatics methods have been widely used for investigating the pathological mechanisms of disease. In this study, datasets including GSE20077 and GSE108724, obtained from the Gene Expression Omnibus (GEO) database, were used for investigating the biomarker and molecular mechanism of HCC. The differentially expressed genes (DEGs) in the datasets were identified, and the targets of the miRNAs were searched in the miRDIP and miRNET databases. Enrichment analysis was performed for delving the molecular mechanism of DEGs, and protein-protein interaction (PPI) networks and miRNA-mRNA networks were used to reveal the hub nodes and the related interaction relationships. Moreover, the expression and diagnostic values of hub nodes were analyzed with the GEPIA2 database. The results showed that 53 upregulated miRNAs and 48 downregulated miRNAs were found in GSE20077, and 55 upregulated miRNAs and 69 downregulated miRNAs were found in GSE108724. Moreover, seven common miRNAs including miR-146b-5p, miR-338-3p, miR-375, miR-502-3p, miR-532-3p, miR-532-5p, and miR-557 were found in the datasets. The targets of the common miRNAs were related with the P53, HIF1, Wnt, and NF-κB pathways. Besides, YWHAZ and CDC42 were identified as the hub nodes and served as the downstream targets of miR-375-3p. The GEPIA2 database showed that YWHAZ and CDC42 were related with the survival rate of the patients. In conclusion, this study suggests that miR-375-3p functions as a tumor suppressor which could inhibit the progression of HCC via targeting YWHAZ and CDC42.

Ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT.

BACKGROUND: Ephrin-A2, a member of the Eph receptor subgroup, is used in diagnosing and determining the prognosis of prostate cancer. However, the role of ephrin-A2 in prostate cancer is remains elusive. METHODS: We established stable clones overexpressing or silencing ephrin-A2 from prostate cancer cells. Then, CCK-8 was used in analyzing the proliferation ability of cells. CD31 staining was used in evaluating angiogenesis. Migration and invasion assay were conducted in vivo and in vitro. The expression of EMT-related markers was evaluated in prostate cancer cells through Western blotting. RESULTS: We revealed that the ectopic expression of ephrin-A2 in prostate cancer cells facilitated cell migration and invasion in vitro and promoted tumor metastasis and angiogenesis in vivo and that the silencing of ephrin-A2 completely reversed this effect. Although ephrin-A2 did not affect tumor cell proliferation in vitro, ephrin-A2 significantly promoted primary tumor growth in vivo. Furthermore, to determine the biological function of ephrin-A2, we assayed the expression of EMT-related markers in stable-established cell lines. Results showed that the overexpression of ephrin-A2 in prostate cancer cells down-regulated the expression of epithelial markers (ZO-1, E-cadherin, and claudin-1) and up-regulated the expression of mesenchymal markers (N-cadherin, ß-catenin, vimentin, Slug, and Snail), but the knocking out of ephrin-A2 opposed the effects on the expression of EMT markers. CONCLUSIONS: These findings indicate that ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT and may be a potentially therapeutic target in metastatic prostate cancer.

Diagnostic value of DACT-2 methylation in serum of prostate cancer patients.

BACKGROUND: Currently, prostate cancer (PCa) remains a hard nut to crack for the medical community. Therefore, the identification and development of novel biomarkers that can accurately diagnose disease and predict prognosis are of paramount importance. The objective of this study was to examine the clinical value of DACT-2 promoter methylation in serum of patients with PCa, to discover a potential diagnostic marker for PCa. METHODS: We investigated the methylation status of DACT-2 in the serum of 64 patients with PCa, 22 patients with benign prostatic hyperplasia (BPH), and 47 healthy subjects by methylation-specific PCR (MSP) and real-time methylation-specific PCR (QMSP). Further, we evaluated the relationship between DACT-2 methylation and clinic pathological parameters. Receiver operating characteristic (ROC) curve analysis was applied to assess the sensitivity, specificity, and diagnostic value of DACT-2 methylation and PSA levels. RESULTS: The results of MSP and QMSP showed that the level of methylation of DACT-2 promoter in patients with PCa was significantly higher than that in patients with BPH and healthy subjects. The PCa patients Gleason score and tumor node metastasis (TNM) positively correlated with promoter methylation level of serum DACT-2. The DACT-2 methylation rate was 0.745 with a sensitivity of 81.8%, and a specificity of 75.0%, the sensitivity, and specificity of PSA was 80.1% and 59.4%. ROC curve results displayed that the diagnostic value of DACT-2 is superior to PSA. CONCLUSIONS: Our study confirms that the level of methylation of the DACT-2 promoter in patients with PCa is much higher than that in patients with benign prostatic hyperplasia (BPH) and healthy subjects, suggesting that DACT-2 methylation in serum is a potential biomarker of PCa.

Plasma Exosomal Long Noncoding RNA lnc-SLC2A12-10:1 as a Novel Diagnostic Biomarker for Gastric Cancer.

PURPOSE: Exosomes participate in cellular communications by transmitting active molecules, including long noncoding RNAs (lncRNAs) and are regarded as suitable candidates for disease diagnosis. This study aimed to identify gastric cancer (GC)-specific exosomal lncRNA and investigate the potential diagnostic value of plasma exosomal lncRNA in GC. PATIENTS AND METHODS: Exosomes from the culture media (CM) of four GC cells (GCCs) and human gastric epithelial cells were isolated. Exosomal RNA was extracted, and lncRNA microarray assay was performed to identify GC-specific exosomal lncRNAs. The expression levels of the candidate exosomal lncRNAs were validated in 120 subjects via quantitative reverse transcription PCR (qRT-PCR). The receiver operating characteristic (ROC) curve and area under curve were used to estimate the diagnostic capacity. We investigated the potential relationship between plasma exosomal lncRNA expression and the clinicopathological parameters of GC. RESULTS: A total of 199 exosomal lncRNAs were expressed at considerable higher levels in GCCs than those in normal controls, among which the top 10 upregulated lncRNAs were selected for further validation in cell, CM, and plasma. qRT-PCR revealed that lnc-SLC2A12-10:1 was remarkably upregulated in exosomes derived from patients with GC and GCCs. The area under the ROC curve was 0.776, which was higher than the diagnostic accuracies of CEA, CA 19-9, and CA72-4. The expression level of exosomal lnc-SLC2A12-10:1 was also significantly correlated with tumor size, TNM stage, lymph node metastasis, and degree of differentiation. The postoperative expression levels of exosomal lnc-SLC2A12-10:1 were lower compared with those of preoperative levels. CONCLUSION: Our study suggested that exosomal lnc-SLC2A12-10:1 may be a potential noninvasive biomarker for the diagnosis and prognosis monitoring of GC. Further large-scale studies are necessary to validate its performance in GC progression.

Knockdown of circRNA circ_0087378 Represses the Tumorigenesis and Progression of Esophageal Squamous Cell Carcinoma Through Modulating the miR-140-3p/E2F3 Axis.

BACKGROUND: We aimed to investigate the function and underlying mechanisms of circ_0087378 in esophageal squamous cell carcinoma (ESCC). METHODS: We verified higher circ_0087378 expression in ESCC tissues by performing qRT-PCR assays. We further confirmed the oncogenic roles of circ_0087378 in ESCC cells through a series of biological function assays. Then, we used an RNA pull-down assay and luciferase reporter assay to identify miR-140-3p that directly interacts with circ_0087378. Subsequent studies were performed to demonstrate that the circ_0087378/miR-140-3p/E2F3 axis promotes ESCC development. RESULTS: We demonstrated that upregulated circ_0087378 expression was positively associated with tumor size, histological grade, tumor stage, the presence of metastasis, and worse survival in patients with ESCC. Our results further revealed that knockdown of circ_0087378 suppressed the proliferation, migration, and invasion of ESCC cells and reduced tumor growth in vivo. Mechanistically, we showed that circ_0087378 could directly bind to miR-miR-140-3p and relieve the suppression for target E2F3, which accelerated cell proliferation, migration, and invasion. Correlation analysis in ESCC specimens supported the involvement of the circ_0087378/miR-140-3p/E2F3 axis in ESCC progression. CONCLUSIONS: This study demonstrated that circ_0087378 might act as a competing endogenous RNA for miR-140-3p, which could inhibit the tumorigenesis and progression of ESCC through upregulating E2F3 expression.

Exosomal long noncoding RNA lnc-GNAQ-6:1 may serve as a diagnostic marker for gastric cancer.

BACKGROUND: Gastric cancer (GC) is one of the most aggressive cancers, with limited early diagnostic measures. Tumor-originated exosomal molecules are regarded as suitable candidates for non-invasive diagnosis. This study aimed to investigate the capacity of exosomal long noncoding RNA lnc-GNAQ-6:1 as a biomarker for early diagnosis of GC. METHODS: In this study, we collected sera from 43 patients with gastric cancer and 27 healthy subjects, then exosomes were isolated using commercial kits. Particle size analysis, Western bloting and protein-based exosomes quantification were conducted to identify the isolated exosomes and to evaluate its yield and purity. Expression levels of exosomal lnc-GNAQ-6:1 were detected by quantitative reverse transcription PCR (qRT-PCR). The serum concentrations of traditional biomarker (CA72-4, CEA, and CA19-9) were measured via a chemiluminescent detection system.The receiver operating characteristic curve (ROC) and area under curve (AUC) were used to estimate the diagnostic capacity. Furthermore, we analyzed the potential relationship between serum exosomal lnc-GNAQ-6:1 expression and clinicopathological parameters of gastric cancer. RESULTS: The exosomes extracted in this study exhibited the typical exosome characteristics and purity. Patients with gastric cancer had the higher exosome yield than healthy volunteer. The results of qRT-PCR showed that compared with the healthy control, the expression of lnc-GNAQ-6:1 was significantly lower in the gastric cancer group. The area under the ROC curve is 0.732, which was higher than the diagnostic accuracy of CEA, CA 19-9 and CA72-4. However, the expression level of lnc-GNAQ-6:1 was not correlated with gender, age, tumor metastasis, serum carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), and carbohydrate antigen 72-4(CA72-4). CONCLUSIONS: Our data demonstrated that serum exosomal lnc-GNAQ-6:1 is lowly expressed in patient with gastric cancer and might be evaluated in larger studies as a new diagnostic marker for gastric cancer.

Serum Exosomal Long Noncoding RNA pcsk2-2:1 As A Potential Novel Diagnostic Biomarker For Gastric Cancer.

PURPOSE: Exosome-shuttled bioactive long non-coding RNA, as novel non-invasive biomarkers for cancer diagnosis, has received increasing attention. Here, we aimed to investigate the expression of serum exosomal long non-coding RNA pcsk2-2:1 (Exo-Lnc RNApcsk2-2:1) in patients of gastric cancer and evaluate its diagnostic value as a marker. PATIENTS AND METHODS: Exosomes were isolated from serum sample of gastric cancer using HiPure Exosomekits and identified via transmission electron microscopy, Western blotting, and nanoparticle tracking analysis. The total exosomal RNA was extracted and reverse transcribed to cDNA. The expression of Exo-Lnc RNA PCSK2-2:1 was detected in serum exosomes of 29 healthy people and 63 gastric cancer patients by real-time quantitative reverse transcription PCR (qRT-PCR), and the relationship between the expression level of Exo-Lnc RNA PCSK2-2:1 and clinicopathological parameters of patients was analyzed. Finally, a receiver operating characteristic curve was used to evaluate the clinical value of Exo-Lnc RNA PCSK2-2:1 as an auxiliary diagnostic marker for gastric cancer. RESULTS: Transmission electron microscopy, nanoparticle size analysis, and Western blotting showed successful separation of serum exosomes. qRT-PCR results revealed that compared with the healthy control, Lnc RNA PCSK2-2:1 expression level in serum exosomes of gastric cancer patients was significantly downregulated (p=0.006). Moreover, the expression level of Exo-Lnc RNA PCSK2-2:1 was correlated with tumor size (p=0.0441), tumor stage (p=0.0061), and venous invasion (p=0.0367). The area under the curve of Exo-Lnc RNA PCSK2-2:1 was 0.896. At the optimal cut-off value, the diagnostic sensitivity and specificity were 84% and 86.5%, respectively. CONCLUSION: Our data indicate that Exo-Lnc RNA PCSK2-2:1 may perform a vital role in the progression of gastric cancer and can be used as a potential marker for the diagnosis of gastric cancer.

Downregulation of DACT-2 by Promoter Methylation and its Clinicopathological Significance in Prostate Cancer.

Backgrounds: Dapper homolog (DACT) 2, a member of DACT gene family, is frequently down-regulated in various malignancies and linked to tumor progression. However, the regulatory mechanism of DACT-2 expression and its biological role in human prostate cancer (PCa) remains elusive. Here, we investigated the expression and an epigenetic change of DACT-2 in prostate cancer, and determined if these findings were correlated with clinicopathologic characteristics of PCa. Methods: The expression profile of DACT-2 of was detected by qRT-PCR, Western blotting, and immunohistochemistry in four prostate cell lines (RWPE-1, LNCaP, PC-3 and DU145), 56 cases of frozen prostate tissues (forty-seven primary prostate carcinomas, nine paired noncancerous and cancerous prostate tissues) and a tissue microarray sets including 100 paraffin-embedded prostate samples (3 normal tissues, 2 cases of adjacent tissues and 95 cases of cancer). Subsequently, the regulatory mechanism of DACT-2 down-regulation was investigated through methylation-specific PCR (MSP) and bisulfite sequencing (BSP). The role of DACT-2 in prostate cancer cell migration and invasion was respectively examined by wound healing and transwell assay. After 5-aza-2'-deoxycytidine treatment of prostate cancer cells, qRT-PCR was used to detect whether the expression of DACT-2 gene mRNA in the cells recovered. Results: Immunohistochemical results shown that the DACT-2 protein was strongly (3+) expressed in the cytoplasm of all 5 noncancerous tissues and 12.7% (12/95) prostate cancer (PCa) tissues. Whereas 68.4% (65/95) PCa samples and 18.9% (18/95) PCa tissues respectively displayed weakly (1+) expressed and moderately (2+) expressed. In addition, DACT-2 expression was negatively associated with Gleason score in tumor specimens (p=0.029). What's more, down-regulation and promoter methylation of DACT-2 were observed in 68.1% (32/47) frozen PCa tissues and all three prostate cancer cell lines. And, the expression of DACT-2 mRNA was restored by the treatment of demethylated drug 5-aza-2'-deoxycytidine in all prostate cancer lines. Prostate cancer cells invasion and migration were significantly suppressed by ectopic expression of DACT-2 in vitro. Conclusions: Our study provides evidence that DACT-2 may be a useful biomarker for distinguishing prostate tumor tissues from non-cancerous samples and a potential target for epigenetic silencing in primary prostate Cancer.

Combined treatment of ABT199 and irinotecan suppresses KRAS-mutant lung cancer cells.

Lung cancer has become the most prevalent neoplasm throughout the world with 1,2 million deaths per year. Molecular genetic analyses have suggested that KRAS mutation is much more frequency in NSCLC. Significant challenges are to develop selective pharmacological inhibitors for RAS mutation to treat cancers driven. In our study, we used the combinatorial strategy to target oncogene addiction for RAS-mutant cells and the data showed that ABT199 and irinotecan leads to RAS-mutant lung cancer cell growth inhibition and enhanced apoptosis in vitro and in vivo. Furthermore, PI3K/AKT signaling was down-regulated by the combination in KRAS-mutant lung cancer cells. Importantly, the effects of ABT199 and irinotecan combination are synergistic on the RAS-mutant lung cancer cells. Therefore, the combination suggests a strong synergy in vivo and a potential avenue for therapeutic treatment of KRAS-mutant cancers which are otherwise difficult targeted by small molecules.

Exosomal ephrinA2 derived from serum as a potential biomarker for prostate cancer.

Up-regulation of serum ephrinA2 is common in various malignancies and has been suggested as a potential biomarker for the diagnosis and prognosis of prostate cancer (PCa). However, the type of serum ephrinA2 expressed in PCa patients remains elusive. Furthermore, the level of exosomal ephrinA2 derived from serum is increased in patients with osteoporosis, a common complication of PCa patients undergoing androgen deprivation therapy. It is unknown whether exosomes derived from PCa patient serum contains ephrinA2. In this study, we explored the ephrinA2 expression in whole serum and tissues and identified the circulating exosomal ephrinA2 as a potential biomarker for PCa. Exosomes were isolated from patient sera by differential centrifugation and the presence of ephrinA2 was confirmed via electron microscopy and western blotting. The type of ephrinA2 in serum was evaluated by western blotting. The expression of serum ephrinA2 including secreted and cleaved ephrinA2 and exosomal ephrinA2 were detected by ELISA and western blotting. Compared with benign prostatic hyperplasia (BPH) and controls, the levels of whole serum ephrinA2 and exosomal ephrinA2 were significantly higher in PCa patients. Moreover, exosomal ephrinA2 expression was positively correlated with TNM staging and Gleason score of PCa patients. The diagnostic efficiency of exosomal ephrinA2 was superior to that of whole serum ephrinA2 and serum PSA in distinguishing PCa patients from those from BPH patents. Our study indicates that exosomal ephrinA2 has high potential as a biomarker for the presence of PCa and offers a new therapeutic target for this disease.

Identification of CSF biomarkers by proteomics in Guillain-Barré syndrome.

The purpose of the present study was to screen for differentially expressed proteins in the cerebrospinal fluid (CSF) of patients with Guillain-Barré syndrome (GBS). The identification of differentially expressed protein can provide new targets for understanding the pathogenic mechanism, early clinical diagnosis, prognosis and for measuring the effectiveness of interventions. We enrolled 50 GBS patients and 50 meningitis patients (control group) to compare protein expression in CSF. The GBS cases included 28 cases of acute inflammatory demyelinating polyneuropathy (AIDP) and 22 cases of acute motor axonal neuropathy (AMAN). We then performed two-dimensional differential in-gel electrophoresis combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to identify the differentially expressed proteins. The expression levels were validated by ELISA, and their accuracy, sensitivity, and specificity in GBS diagnosis were analyzed by the receiver operating characteristic curve. Three differentially expressed proteins were identified, including the upregulated haptoglobin (Hp) and heat shock protein 70 (Hsp70), and downregulated cystatin C. There were no significant differences between the AIDP and AMAN patients in the positive rates and quantitative expression levels of the three differentially expressed proteins. The accuracy of Hp in the diagnosis of GBS was 0.835, sensitivity was 86.7%, and specificity was 88.2%. The accuracy of cystatin C in the diagnosis of GBS was 0.827, sensitivity was 85.5%, and specificity was 89.7%. The accuracy of Hsp70 in the diagnosis of GBS was 0.841, its sensitivity was 87.8%, and its specificity was 92.3%. Hp and Hsp70 are significantly increased, and cystatin C is downregulated in CSF of GBS patients, which provides important biomarkers for early GBS diagnosis, although these proteins cannot distinguish AIDP and AMAN.

Eph/ephrin family anchored on exosome facilitate communications between cells.

Interactions of Ephrins and Eph receptors at cell membranes play crucial role in boundary formation and axon guidance. Extracellular vesicles (EVs), such as exosomes, are formed by cells communicating with each other in paracrine or endocrine manner. Until now, it is thought that direct cell-cell contact is necessary for ephrin and Eph receptor signal transduction. In this review, we discuss recent data that indicate the existence of a novel Eph-ephrin family anchored exosome signaling pathway in long-range intercellular communication and provide evidence that this type of signaling elicits cellular responses in cancer cells, independent of juxtacrine interactions. We emphasize that exosome-anchored Eph/ephrin involves a variety of biological processes and transduction signals, which may serve as a potential diagnostic biomarker.