Risk Factors of Lymph Node Metastasis and Prognosis in 891 Chinese Patients With Submucosal Early Gastric Carcinoma, Emphasizing Differences Between Gastric Cardiac and Noncardiac Origins.

Differences in risk factors (RF) of lymph node metastasis (LNM) and prognosis between submucosal early gastric cardiac (SEGCC) and noncardiac (SEGNCC) carcinomas remain unclear. In this study, we investigated and compared RF of LNM and prognosis in 891 patients with radical gastrectomy for SEGCC (n=217) or SEGNCC (n=674). Compared with SEGNCC, SEGCC displayed significantly higher proportion of elderly patients (70 y or above), the elevated macroscopic type, well/moderately differentiated tubular and low-grade papillary adenocarcinomas, as well as low-grade tumor budding, but lower prevalence of the depressed macroscopic type, poorly differentiated tubular adenocarcinoma, mixed adenocarcinoma, poorly cohesive carcinoma, lymphovascular invasion (LVI), perineural invasion, and high-grade tumor budding. By univariate analysis, significant RF for LNM of the cohort included female sex, poor differentiation, SM2 invasion, LVI, intermediate-grade and high-grade tumor budding, whereas tumor size, histology type, and perineural invasion were the significant RF for LNM in SEGNCC. By multivariate analysis, significant independent RF for LNM included female sex and LVI in SEGCC but were female sex, mixed adenocarcinoma, LVI, and high-grade tumor budding in SEGNCC. The 5-year overall survival was significantly worse in SEGCC than in SEGNCC for patients with LNM, but not for those without. For overall survival, LNM was the only significant independent RF in SEGCC, whereas age 70 years or above and LNM were independent RF in SEGNCC. The results of our study provided the clinicopathologic evidence for individualized clinical management strategies for these 2 groups of patients and suggested different pathogenesis mechanisms between SEGCC and SEGNCC.

Clear cell hepatocellular carcinoma: Gd-EOB-DTPA-enhanced MR imaging features and prognosis.

PURPOSE: To investigate imaging findings on gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging (Gd-EOB-DTPA-enhanced MRI) and prognosis of clear cell hepatocellular carcinoma (CCHCC) comparing with non-otherwise specified hepatocellular carcinoma (NOS-HCC). METHODS: The clinical, pathological and MR imaging features of 42 patients with CCHCC and 84 age-matched patients with NOS-HCC were retrospectively analyzed from January 2015 to October 2021. Univariate and multivariate logistic regression and Cox regression analyses were performed to identify independent diagnostic and prognostic factors for CCHCC. Disease-free survival (DFS) and overall survival (OS) were determined by Kaplan-Meier analysis. RESULTS: CCHCC showed fat content more frequently (P < 0.001) and relatively higher Edmondson tumor grade (P = 0.001) compared with NOS-HCC. The lesion-to-muscle ratio (LMR) and lesion-to-liver ratio (LLR) of CCHCC on pre-enhancement T1-weighted imaging (pre-T1WI) (P = 0.001, P = 0.003) and hepatobiliary phase (HBP) (P = 0.007, P = 0.048) were significantly higher than those of NOS-HCC. The area under the curve (AUC) for fat content, LLR on pre-T1WI and their combination with better diagnostic performance in predicting CCHCC were 0.678, 0.666, and 0.750, respectively. There was no statistically significant difference in clinical outcomes between CCHCC and NOS-HCC. Multivariate Cox analysis confirmed that tumor size > 2 cm and enhancing capsule were independent prognostic factors for DFS and OS among CCHCC patients. CONCLUSION: Fat content and adjusted lesion signal intensity on pre-T1WI and HBP could be used to differentiate CCHCC from NOS-HCC. CCHCC had similar prognosis with NOS-HCC.

Hsa_circ_0046430 promotes the progression of colorectal cancer by targeting miR-6785-5p/SRCIN1 axis as a ceRNA.

The correlation among circular RNAs (circRNAs), microRNAs, and messenger RNAs have gained increasing attention in recent years. However, the mechanism of such discoveries in colorectal cancer (CRC) is not yet elucidated. The present study aimed to clarify whether the novel circRNAs regulate the prognosis-related genes through the competing endogenous RNAs (ceRNA). An analysis of the Weighted Gene Co-Expression Network Analysis was conducted to screen a module-trait circRNAs, and other big data mining technologies were used to predict the related microRNAs and the downstream genes. Prognosis-related gene model was built using the Cox regression analysis for the 138 messenger RNAs associated with hsa circ 0046430. The qRT-PCR was adopted to verify ceRNA network. Immunohistochemistry verified the correlation between SRCIN1 and patient prognosis. In summary, these results demonstrated that hsa_circ_0046430 is a tumor-related circRNA based on the clinical characteristics module of Weighted Gene Co-Expression Network Analysis. The prognostic risk score signature model analysis indicated that CRC risk was independently related to the risk score and SRCIN1 was independently associated with overall survival. Therefore, the hsa_circ_0046430/miR-6785-5p/SRCIN1 axis was constructed. Hsa_circ_0046430/miR-6785-5p/SRCIN1 axis relative expression level was determined by qRT-PCR. Immunohistochemical staining further validated that SCRIN1 was significantly higher in cancer than in adjacent normal tissues. Our study identified and primarily validated the hsa_circ_0046430/miR-6785-5p/SRCIN1 regulatory axis impacted on CRC prognosis, suggesting novel biomarkers and therapeutic targets for CRC patients. Further in-depth studies are essential to confirm the underlying ceRNA mechanism.

Nomogram based on MRI for preoperative prediction of Ki-67 expression in patients with intrahepatic mass cholangiocarcinoma.

OBJECTIVES: To validate a new nomogram based on magnetic resonance imaging (MRI) for pre-operative prediction of Ki-67 expression in patients with intrahepatic mass cholangiocarcinoma (IMCC). METHODS: A total of 78 patients with clinicopathologically confirmed IMCC who underwent pre-operative gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid enhanced MRI between 2016 and 2022 were enrolled in the training and validation group (53 patients and 25 patients, respectively). Images including qualitative, quantitative MRI features and clinical data were evaluated. Univariate analysis and multivariate logistic regression were used to select the independent predictors and establish different predictive models. The predictive performance was validated by operating characteristic curve (ROC) analysis, calibration curve, and decision curve analysis (DCA). The validation cohort was used to test the predictive performance of the optimal model. The nomogram was constructed with the optimal model. RESULTS: In the training cohort, independent predictors obtained from the combined model were DWI (OR 1822.741; 95% CI 6.189, 536,781.805; P = 0.01) and HBP enhancement pattern (OR 14.270; 95% CI 1.044, 195.039; P = 0.046). The combined model showed the good performance (AUC 0.981; 95% CI 0.952, 1.000) for predicting Ki-67 expression. In the validation cohort, The combined model (AUC 0.909; 95% CI 0.787, 1.000)showed the best performance compared to the clinical model (AUC 0.448; 95% CI 0.196, 0.700) and MRI model (AUC 0.770; 95% CI 0.570, 0.970). CONCLUSION: This new nomogram has a good performance in predicting Ki-67 expression in patients with IMCC, which could help the decision-making of the patients' therapy strategies.

Nomogram Estimating Vessels Encapsulating Tumor Clusters in Hepatocellular Carcinoma From Preoperative Gadoxetate Disodium-Enhanced MRI.

BACKGROUND: Vessels encapsulating tumor clusters (VETC) pattern is a novel microvascular pattern associated with poor outcomes of hepatocellular carcinoma (HCC). Preoperative estimation of VETC has potential to improve treatment decisions. PURPOSE: To develop and validate a nomogram based on gadoxetate disodium-enhanced MRI for estimating VETC in HCC and to evaluate whether the estimations are associated with recurrence after hepatic resection. STUDY TYPE: Retrospective. POPULATION: A total of 320 patients with HCC and histopathologic VETC pattern assessment from three centers (development cohort:validation cohort = 173:147). FIELD STRENGTH/SEQUENCE: A3.0  T/turbo spin-echo T2-weighted, spin-echo echo-planar diffusion-weighted, and 3D T1-weighted gradient-echo sequences. ASSESSMENT: A set of previously reported VETC- and/or prognosis-correlated qualitative and quantitative imaging features were assessed. Clinical and imaging variables were compared based on histopathologic VETC status to investigate factors indicating VETC pattern. A regression-based nomogram was then constructed using the significant factors for VETC pattern. The nomogram-estimated VETC stratification was assessed for its association with recurrence. STATISTICAL TESTS: Fisher exact test, t-test or Mann-Whitney test, logistic regression analyses, Harrell's concordance index (C-index), nomogram, Kaplan-Meier curves and log-rank tests. P value < 0.05 was considered statistically significant. RESULTS: Pathological VETC pattern presence was identified in 156 patients (development cohort:validation cohort = 83:73). Tumor size, presence of heterogeneous enhancement with septations or with irregular ring-like structures, and necrosis were significant factors for estimating VETC pattern. The nomogram incorporating these indicators showed good discrimination with a C-index of 0.870 (development cohort) and 0.862 (validation cohort). Significant differences in recurrence rates between the nomogram-estimated high-risk VETC group and low-risk VETC group were found (2-year recurrence rates, 50.7% vs. 30.3% and 49.6% vs. 31.8% in the development and validation cohorts, respectively). DATA CONCLUSION: The nomogram integrating gadoxetate disodium-enhanced MRI features was associated with VETC pattern preoperatively and with postoperative recurrence in patients with HCC. EVIDENCE LEVEL: 4 TECHNICAL EFFICACY: Stage 2.

A Retrospective Cohort Study of Factors Influencing Lymph Node Metastasis in Patients With Early Gastric Papillary Adenocarcinoma.

INTRODUCTION: High risk of lymph node metastasis (LNM) in gastric papillary adenocarcinoma causes endoscopists to worry about the suitability of endoscopic resection for early gastric papillary adenocarcinoma (EPAC). We compared risk factors and attempted to establish a scoring system to stratify LNM risk in patients with EPAC. METHODS: A retrospective analysis was performed on 2,513 patients with early gastric carcinoma (EGC) who underwent radical resection in 4 tertiary hospitals in China. Univariate and multivariate analyses were performed to compare the invasiveness in EPAC and other types of EGC and to evaluate potential factors in predicting LNM risk in EPAC groups. RESULTS: Three hundred thirty-five patients with EPAC were enrolled in our study, of which 62 patients were found to have LNM. After comparing clinicopathological characteristics of EPAC with and without LNM, the following factors were included in the risk scoring system: 1 point each for lower stomach location and tumor size >2.0 cm, 3 points for lymphovascular invasion, and 4 points for submucosal invasion; the risk scoring system was validated in a small internal validation set with an area under the curve of 0.844. DISCUSSION: Our results suggested that EPAC was highly invasive compared with other EGCs, especially differentiated EGC types, and need to be treated more rigorously. This proposed risk scoring system could stratify LNM risk in patients with EPAC, and endoscopic resection may only be performed safely on the groups with a low LNM rate.

Native T1-mapping and diffusion-weighted imaging (DWI) can be used to identify lung cancer pathological types and their correlation with Ki-67 expression.

Background: This study aimed to explore the value of native T1-mapping and diffusion-weighted imaging (DWI) in differentiating the pathological types and degree of tumor differentiation of lung cancer and their correlation with Ki-67 protein expression. Methods: A total of 78 consecutive lung cancer patients who received chest magnetic resonance imaging (MRI) scans between May 2020 and June 2021 were enrolled in this study. Two radiologists independently analyzed the apparent diffusion coefficient (ADC) and T1 values for each lesion. The intraclass correlation coefficient (ICC) and Bland-Altman plots were generated to assess interobserver agreement of the T1 and ADC mean values in lesions. The difference in ADC and T1 values among different pathological types, as well as between high- and low-differentiated lung cancers were analyzed, and diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve analysis. The correlation between ADC value, T1 value, and Ki-67 protein expression index was determined. Results: The ADC and T1 values showed excellent interobserver agreement (ICC 0.820, 0.942, respectively). There was a significant difference in ADC values between small cell carcinoma and squamous carcinoma (P<0.05), and between small cell carcinoma and adenocarcinoma (P<0.05), but not between squamous carcinoma and adenocarcinoma (P>0.05). A significant difference in T1 values was observed between small cell carcinoma (P<0.05) and adenocarcinoma, and between squamous carcinoma (P<0.05) and adenocarcinoma, but not between squamous carcinoma and small cell carcinoma (P>0.05). There were statistically significant differences in ADC and T1 values between the moderately and highly differentiated group and the poorly differentiated group (P<0.05). ROC curve analysis showed that the T1 combined with ADC value had high diagnostic value for the degree of differentiation of the tumor [area under the curve (AUC) =0.912]. Pearson correlation analysis showed a significant positive correlation between T1 value and Ki-67 index (r=0.66, P<0.001) and a significant negative correlation between ADC value and Ki-67 index (r=-0.45, P<0.01). Conclusions: T1 and ADC values can be used to distinguish the pathological type and differentiation degree of lung cancer.

High-grade Papillary Early Gastric Carcinoma With High Risk for Lymph Node Metastasis and Poor Prognosis: A Clinicopathologic Study of 96 Cases Among 1136 Consecutive Radical Gastrectomies.

Papillary early gastric carcinoma (EGC) is believed to have a low risk of lymph node metastasis (LNM) and thus can be resected endoscopically. We observed anecdotally that some papillary EGC tumors showed conspicuous high-grade dysplastic features, but the significance of these observations is unknown. In this bicenter study we investigated papillary EGCs that were divided into high-grade (n=96) and low-grade (n=118) groups among 1136 consecutive EGC radical resection cases. Concurrent 464 well-moderately differentiated tubular EGCs were served as the control group. Compared with low-grade papillary and well-moderately differentiated tubular EGCs, high-grade papillary EGC displayed significantly larger sizes (mean 2.51 cm), higher frequencies of the elevated macroscopic type (51%), lymphovascular invasion (LVI) (38.5%), and LNM (31.2%). Low-grade papillary EGCs exhibited a higher prevalence of the elevated macroscopic type, but not LVI nor LNM, compared with tubular EGC. Independent risk factors for LNM included high-grade histology, female sex, distal location, submucosal invasion, and LVI. The 5-year overall survival rate was significantly lower in high-grade (79.6%) papillary than in low-grade (88.9%) papillary or tubular (92.8%) EGCs, while no significant difference in prognosis was observed in the latter 2 groups. Age of 66 years or older and LNM were independent risk factors for overall survival. In conclusions, high-grade papillary EGC was associated with high frequencies of LVI, LNM, and poor prognosis, and thus unsuitable for endoscopic therapy, while low-grade papillary EGC showed clinicopathologic features and prognosis similar to well-moderately differentiated tubular EGC and may be treated endoscopically in appropriate clinical settings.

Texture Analysis Based on Gd-EOB-DTPA-Enhanced MRI for Identifying Vessels Encapsulating Tumor Clusters (VETC)-Positive Hepatocellular Carcinoma.

PURPOSE: To determine the potential findings associated with vessels encapsulating tumor clusters (VETC)-positive hepatocellular carcinoma (HCC), with particular emphasis on texture analysis based on gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI. METHODS: Eighty-one patients with VETC-negative HCC and 52 patients with VETC-positive HCC who underwent Gd-EOB-DTPA-enhanced MRI before curative partial hepatectomy were retrospectively evaluated in our institution. MRI texture analysis was performed on arterial phase (AP) and hepatobiliary phase (HBP) images. The least absolute shrinkage and selection operator (LASSO) logistic regression was used to select texture features most useful for identifying VETC-positive HCC. Univariate and multivariate analyses were used to determine significant variables for identifying the VETC-positive HCC in clinical factors and the texture features of MRI. Receiver operating characteristic (ROC) analysis and DeLong test were performed to compare the identified performances of significant variables for identifying VETC-positive HCC. RESULTS: LASSO logistic regression selected 3 features in AP and HBP images, respectively. In multivariate analysis, the Log-sigma-4.0-mm-3D first-order Kurtosis derived from AP images (odds ratio [OR] = 4.128, P = 0.001) and the Wavelet-LHL-GLDM Dependence Non Uniformity Normalized derived from HBP images (OR = 2.280, P = 0.004) were independent significant variables associated with VETC-positive HCC. The combination of the two texture features for identifying VETC-positive HCC achieved an AUC value of 0.844 (95% confidence interval CI, 0.777, 0.910) with a sensitivity of 80.8% (95% CI, 70.1%, 91.5%) and specificity of 74.1% (95% CI, 64.5%, 83.6%). CONCLUSION: Texture analysis based on Gd-EOB-DTPA-enhanced MRI can help identify VETC-positive HCC.

Imaging features based on Gd-EOB-DTPA-enhanced MRI for predicting vessels encapsulating tumor clusters (VETC) in patients with hepatocellular carcinoma.

OBJECTIVE: To investigate the non-invasive prediction of hepatocellular carcinoma (HCC) with vessels encapsulating tumor clusters (VETC) based on qualitative and quantitative imaging features of gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI. METHODS: 109 patients with pathologically confirmed HCC who underwent Gd-EOB-DTPA enhanced MRI and immunochemical staining for CD34 were retrospectively evaluated in our institution (the first affiliated hospital of Soochow university). Pre-operative imaging features of Gd-EOB-DTPA-enhanced MRI were qualitatively and quantitatively reviewed by radiologists. Significant variables for differentiating the VETC-positive and VETC-negative HCCs were identified in univariate and multivariate analyses. Receiver operating characteristic (ROC) analysis was performed to determine the optimal cut-off values for quantitative variables. The nomogram based on the coefficient of multivariate analysis was constructed to evaluate the probability of VETC-positive HCCs. RESULTS: The multivariate analysis showed that the serum AST level >40 U l-1 (p = 0.007), non-rim diffuse and heterogeneous arterial phase hyperenhancement (p = 0.035), tumor-to-liver SI ratio of 1.135 or more on AP images (p = 0.001), and tumor-to-liver SI ratio of 0.585 or less on HBP images (p = 0.002) were significant predictors for predicting VETC-positive HCCs. Combing all four significant variables provided a diagnostic accuracy of 82.6%, sensitivity of 83.9%, specificity of 80.9% for identifying VETC status. The area under the receiver operating characteristics curve value of the logistical regression coefficient-based nomogram was 0.885 (95% confidence intervals, 0.824-0.946). CONCLUSION: Qualitative and quantitative imaging features of Gd-EOB-DTPA-enhanced MRI integrating laboratory examination can provide good diagnostic performance. ADVANCES IN KNOWLEDGE: VETC is a novel identified microvascular pattern; associations between imaging features and VETC status have not been investigated. Pre-operative diagnosis of VETC status in HCC is essential to help predict the outcome of patients and make a decision for the therapeutic schedule.

Identification of vital genes and pathways associated with mucosal melanoma in Chinese.

Mucosal melanoma is a rare malignant melanoma with more aggressive and poorer outcomes. The incidence of mucosal melanoma varies greatly among different ethnic groups. We herein sought to characterize the vital genes and pathways of Chinese mucosal melanoma patients. By whole-exome sequencing in six patients with mucosal melanoma, we detected a total of 21,733 CNVs and 2372 SNPs. The CNV/SNP burden varies greatly between individuals, including recurrent CNV targeting PIK3 family, KRAS, APC and BRCA1. Significantly mutated genes were NUDT5, ZBTB18, NEURL4, ZNF430, RBM44, GAK, PCDHA13, STK38 and UBR5. Besides, FAT1 gene was identified frequently mutated in anorectal melanoma patients (3/3, 100%). Moreover, our result showed that HPV infection may be associated with mucosal melanoma. In conclusion, this study indicated that mucosal melanomas have a low SNPs burden and a high number of CNVs and expand the spectrum of mucosal melanoma variants, also provided an insight for the pathological mechanism of mucosal melanoma.

Tumor Budding and Other Risk Factors of Lymph Node Metastasis in Submucosal Early Gastric Carcinoma: A Multicenter Clinicopathologic Study in 621 Radical Gastrectomies of Chinese Patients.

Compared with early gastric intramucosal carcinoma, submucosal carcinoma is known to have a higher risk for lymph node metastasis (LMN), but risk factors in submucosal carcinoma remain elusive. In this multicenter study with 621 radical gastrectomies for submucosal early gastric carcinoma, we investigated tumor budding and other risk factors of LMN that were identified in 172 cases (27.7%). Overall, independent high-risk factors for LMN included lymphovascular invasion (odds ratio, 3.9; 95% confidence interval, 2.5-6.1), tumor budding (odds ratio, 3.3; 95% confidence interval, 1.9-5.9), mixed tubular/papillary adenocarcinoma with poorly cohesive carcinoma (odds ratio, 2.1; 95% confidence interval, 1.0-4.3), and female sex (odds ratio, 1.6; 95% confidence interval, 1.0-2.6), whereas gastric cardiac submucosal carcinomas had a significantly lower risk for LMN (odds ratio, 0.5; 95% confidence interval, 0.3-0.9). In 276 well/moderately differentiated tubular or papillary submucosal early gastric carcinomas, independent risk factors were tumor budding (odds ratio, 3.7; 95% confidence interval, 1.6-8.7), deep submucosal (SM2) invasion (odds ratio, 3.1; 95% confidence interval, 1.3-7.6), and lymphovascular invasion (odds ratio, 2.7; 95% confidence interval, 1.3-5.6). In 174 cases without tumor budding and lymphovascular invasion, no LMN was identified in 47 cardiac tumors, and 15 tumors <1.0 cm in size. In conclusion, tumor budding, lymphovascular invasion, mixed tubular/papillary adenocarcinoma with poorly cohesive carcinoma, and female gender were found to be significant high-risk factors for LMN in submucosal early gastric carcinoma, while submucosal gastric cardiac carcinoma had a significantly lower risk for nodal metastasis.

Micropapillary early gastric carcinoma with distinct clinicopathological features, high risk for lymph node metastasis, and dismal prognosis: a multicenter clinicopathological study of 29 cases identified in 1890 early gastric carcinoma radical gastrectomies.

Clinicopathology and risk factors of lymph node metastasis (LNM) in micropapillary early (pT1) gastric carcinoma (MEGC) remain elusive because of the extreme rarity. In this multicenter study, we investigated 1890 consecutive radical resections of early gastric carcinoma diagnosed with the World Health Organization criteria and identified 29 (1.5%) MEGC cases with a small (≥5%) micropapillary component. MEGC showed a male predominance (male-to-female ratio, 21:8). Most (93.1%; 27/29) tumors invaded submucosa. Lymphovascular invasion was detected in 14 (48.3%) of 29 cases. LNM was found in 13 cases (44.8%; 11 identified with a routine hematoxylin-eosin stain and 2 additional cases with a positive pancytokeratin immunostain). Overall, independent risk factors for LNM in early gastric carcinoma included patient age of 62 years or less, female sex, noncardiac location, ulcerative pattern, tumor size of greater than 2 cm, submucosal invasion, Lauren diffuse type, lymphovascular invasion, and MEGC. In MEGC, advanced pathologic stages were demonstrated in 6 (20.7%) of 29 cases. The 5-year overall survival rate of MEGC patients was 58.6%. Submucosal invasion, lymphovascular invasion, and LNM were significantly more frequent in the MEGC group than in the non-MEGC groups. Advanced pathologic stages were significantly more common in MEGC than in nonmicropapillary Lauren intestinal- but not diffuse-type early gastric carcinomas. In conclusion, MEGC demonstrated a high propensity for lymphovascular invasion, LNM with advanced stages, and dismal prognosis.

Eukaryotic Elongation Factor 2 (eEF2) is a Potential Biomarker of Prostate Cancer.

Eukaryotic elongation factor 2 (eEF2), a key regulator of protein synthesis, is involved in the progression of several types of cancer. This first study was to investigate the relationships between eEF2 protein and prostate cancer (PCa). Immunohistochemical staining was used to verify eEF2 protein in a set of 97 formalin-fixed, paraffin-embedded primary PCa tissues. Expression of eEF2 protein in positive cells was characterized by cytoplasmic staining. Correlations with clinicopathological factors were evaluated by Chi-square or Fisher's exact probability tests. eEF2 protein was found in 74 out of 97 (76.29%) patients. eEF2-positive had higher PSA and Gleason score than negative in all patients. In addition, the positive expression of eEF2 protein was significantly associated with PSA and Gleason score (P = 0.007 and 0.002). However, no significant correlations occurred between expression of eEF2 protein and TNM stage (P = 0.292). In those eEF2 protein-positive patients, we have found staining intensity of eEF2 protein was not only associated with PSA and Gleason score, but also associated with TNM stage (P = 0, 0.014 and 0.001, respectively). To conclude, our study indicates that expression of eEF2 protein is a potential biomarker for evaluating PCa.

The long non-coding RNA NONHSAT062994 inhibits colorectal cancer by inactivating Akt signaling.

The aberrant expression of long noncoding RNAs (lncRNAs) is implicated in cancer development and progression. However, the clinical significance and mechanism by which NONHSAT062994 regulates colorectal cancer (CRC) is unknown. We here reported that NONHSAT062994 was significantly downregulated in human CRC tissues and cell lines. Moreover, its expression was inversely correlated with tumor size and overall survival (OS) time in CRC patients. In CRC cells, the overexpression and knockdown of NONHSAT062994 inhibited and enhanced CRC cell growth, respectively, in vitro and in vivo. Mechanistically, NONHSAT062994 functioned as a tumor suppressor to inhibit CRC cell growth by inactivating Akt signaling. Notably, the NONHSAT062994 expression status was negatively correlated with the Akt downstream targets c-Myc and Cyclin D1 in clinical CRC samples. The current findings suggest that NONHSAT062994 plays a critical role in the development of CRC by regulating Akt signaling, and identified a candidate prognostic biomarker or potential therapeutic target for CRC patients.

Prostaglandin E2 promotes human cholangiocarcinoma cell proliferation, migration and invasion through the upregulation of ß-catenin expression via EP3-4 receptor.

Prostaglandin E2 (PGE2) is involved in cholangiocarcinoma cell proliferation, migration and invasion through E prostanoid receptors, including EP1, EP2 and EP4. However, the functions and the mechanisms of those splice variants of EP3 receptors in promoting liver cancer cell growth and invasion remain to be elucidated. In our previous studies, four isoforms of EP3 receptors, EP3-4, EP3-5, EP3-6 and EP3-7 receptors, were detected in CCLP1 and HuCCT1 cells. However, the functions of these receptors in these cells have yet to be determined. It was reported that ß-catenin is closely correlated with malignancy, including cholangiocarcinoma. The present study was designed to examine the effects of 4-7 isoforms of EP3 in promoting cholangiocarcinoma progression and the mechanisms by which PGE2 increases ß-catenin protein via EP3 receptors. The results showed that PGE2 promotes cholangiocarcinoma progression via the upregulation of ß-catenin protein, and the EP3-4 receptor pathway is mainly responsible for this regulation. These findings reveal that PGE2 upregulated the cholangiocarcinoma cell ß-catenin protein through the EP3-4R/Src/EGFR/PI3K/AKT/GSK-3ß pathway. The present study identified the functions of EP3 and the mechanisms by which PGE2 regulates ß-catenin expression and promoted cholangiocarcinoma cell growth and invasion.

Prostaglandin E2 promotes the cell growth and invasive ability of hepatocellular carcinoma cells by upregulating c-Myc expression via EP4 receptor and the PKA signaling pathway.

Hepatocellular carcinoma (HCC) represents a major health problem worldwide. Prostaglandin E2 (PGE2), the predominant product of cyclooxygenase-2, has been implicated in hepatocarcinogenesis. However, the underlying molecular mechanisms remain to be further elucidated. c-myc, a cellular proto-oncogene, is activated or overexpressed in many types of human cancer, including HCC. The present study was designed to investigate the internal relationship and molecular mechanisms between PGE2 and c-Myc in HCC, and to define its role in HCC cell growth and invasion. Our results showed that PGE2 significantly upregulated c-Myc expression at both the mRNA and protein levels, and knockdown of c-Myc blocked PGE2-induced HCC cell growth and invasive ability in human HCC Huh-7 cells. The effect of PGE2 on c-Myc expression was mainly through the EP4 receptor, and EP4 receptor-mediated c-Myc protein upregulation largely depended on de novo biosynthesis of c-Myc mRNA and its protein. EP4 receptor signaling activated GS/AC and increased the intracellular cAMP level in Huh-7 cells. The adenylate cyclase (AC) activator forskolin mimicked the effects of the EP4 receptor agonist on c-Myc expression, while the AC inhibitor SQ22536 reduced EP4 receptor-mediated c-Myc upregulation. These data confirm the involvement of the GS/AC/cAMP pathway in EP4 receptor-mediated c-Myc upregulation. Moreover, the phosphorylation levels of CREB protein were markedly elevated by EP4 receptor signaling, and by using specific inhibitor and siRNA interference, we demonstrated that PKA/CREB was also involved in the EP4 receptor-mediated c-Myc upregulation. In summary, the present study revealed that PGE2 significantly upregulates c-Myc expression at both mRNA and protein levels through the EP4R/GS/AC/cAMP/PKA/CREB signaling pathway, thus promoting cell growth and invasion in HCC cells. Targeting of the PGE2/EP4R/c-Myc pathway may be a new therapeutic strategy to prevent and cure human HCC.

Prostaglandin E2 accelerates invasion by upregulating Snail in hepatocellular carcinoma cells.

Our previous studies showed that prostaglandin E2 (PGE2) promotes hepatoma cell growth and migration, as well as invasion; however, the precise mechanism remains elusive. Snail and p65 protein levels were detected in human samples with hepatocellular carcinoma (HCC) by immunohistochemistry (IHC) staining. HCC cell lines (Huh-7 and Hep3B) were used for in vitro experiments. PGE2/Akt/NF-κB pathway was investigated in Huh-7 and Hep3B cells after treatment with PGE2, EP4 receptor (EP4R) agonist, Akt inhibitor, and NF-κB inhibitor, respectively, by real-time reverse transcription (RT)-PCR, Western blotting, and immunofluorescence (IF) staining. In vitro cell invasion assay was performed to evaluate the effect of PGE2 on tumor invasiveness. Knockdown of EP4R was carried out in Huh-7 cells through plasmid-based small interfering RNA (siRNA) approach to confirm the regulation of PGE2 on Snail by EP4R. Dual luciferase reporter assay was performed to assess Snail promoter activity in Huh-7 cell after treatment with EP4R agonist. We found that the protein levels of Snail were higher in HCC tissues than those in control and that PGE2 and EP4R agonist treatment significantly increased Snail expression in Huh-7 and Hep3B cells. EP4R agonist also profoundly promoted invasiveness of Huh-7 cells. Knockdown of the EP4R by siRNA completely blocked the PGE2-induced upregulation of Snail expression and reduced invasiveness of Huh-7 cells. We failed to find that EP4R-induced upregulation of Snail was reversed by inhibition of cAMP response element-binding protein (CREB), a canonical downstream target of EP4R. Alternatively, EP4R agonist treatment significantly increased the levels of phosphorylated EGFR and Akt both in Huh-7 and Hep3B cells. AG1478, an EGFR inhibitor, blocked the phosphorylation of Akt. The levels of phosphorylated IκB increased in Huh-7 cells after treatment with EP4R agonist for 30 min. The levels of phosphorylated p65 started to increase in Huh-7 cells treated with EP4R agonist for 4 h, and p65 translocated into the nucleus. In EP4R-agonist-treated Hep3B, the levels of phosphorylated p65 were also increased compared to the control group. The phosphorylation levels of p65 were significantly decreased in Huh-7 and Hep3B cells after treatment with the Akt signaling inhibitor LY294002 and EP4R agonist for 24 h. Treatment with the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) at 10 µM for 24 h blocked EP4R-agonist-induced Snail upregulation in Huh-7 and Hep3B cells. Furthermore, we obtained human Snail promoter sequence from TRED-Promoter Database and identified a putative binding site of NF-κB in the sequence through TFSEARCH analysis. Subsequently, we treated Huh-7 cells with EP4R agonist or EP4R agonist and PDTC (NF-κB antagonist) and found significantly increased Snail promoter activity after EP4R agonist treatment for 12 h. The increased Snail promoter activity could be partially abolished by additional PDTC treatment. In addition, p65 protein levels were found increased together with Snail in HCC tissues compared to normal liver tissues. In conclusion, PGE2 activates Akt/NF-κB signaling and then upregulates Snail via the EP4R/EGFR to promote migration and invasion in hepatoma cells. These findings may help future evaluation of novel chemo-preventive strategies for HCC.

Prostaglandin E2 receptor EP2 mediates Snail expression in hepatocellular carcinoma cells.

Prostaglandin E2 (PGE2) has been shown to influence cell invasion and metastasis in several types of cancer, including hepatocellular carcinoma (HCC). however, the molecular mechanisms underlying it remain to be further elucidated. Snail, as one of key inducers of epithelial-mesenchymal transition (EMT), plays pivotal roles in HCC invasion and metastasis. The present study was designed to evaluate the possible signaling pathways through which PGE2 regulates Snail protein expression in HCC cell lines. PGE2 markedly enhanced Huh-7 cell invasion and migration ability by upregulating the expression level of Snail protein, and EP2 receptor played an important role in this process. Src, EGFR, Akt and mTOR were all activated and involved in the regulation of snail protein expression. Our findings suggest that PGE2 could upregulate the expression level of Snail protein through the EP2/Src/EGFR/Akt/mTOR pathway in Huh-7 cells, which promotes HCC cell invasion and migration.

Prostaglandin E2 promotes hepatocellular carcinoma cell invasion through upregulation of YB-1 protein expression.

Prostaglandin E2 (PGE2) has been implicated in hepatocellular carcinoma cell invasion. Recently, it was reported that Y box-binding protein 1 (YB-1) is closely correlated with malignancy. This study was designed to examine the mechanisms by which PGE2 increases YB-1 expression and promotes HCC cell invasion. PGE2 greatly enhanced HCC cell invasion through upregulation of the YB-1 protein, and the EP1 receptor is mainly responsible for this regulation. Src and EGFR were both activated by PGE2, which in turn increased the phosphorylation levels of p44/42 MAPK. Src, EGFR and p44/42 MAPK were all involved in PGE2-induced YB-1 expression. Chemical inhibitors and RNAi analysis all confirmed the role of mTOR complex 1 in YB-1 expression induced by PGE2. Furthermore, YB-1 was able to regulate the expression of a series of EMT-associated genes, which indicated that YB-1 could have the potential to control the epithelial-mesenchymal transition process in HCC cells. These findings reveal that PGE2 upregulated YB-1 expression through the EP1/Src/EGFR/p44/42 MAPK/mTOR pathway, which greatly enhanced HCC cell invasion. This study for the first time describes the mechanisms through which PGE2 regulates YB-1 expression and promotes HCC cell invasion.