Development and validation of a novel biomarker panel for Crohn's disease and rheumatoid arthritis diagnosis and treatment.

BACKGROUND: Crohn's disease (CD) and rheumatoid arthritis (RA) are immune-mediated inflammatory diseases. However, the molecular mechanisms linking these two diseases remain unclear. METHODS: To identify shared core genes between CD and RA, we employed differential gene analysis and the least absolute shrinkage and selection operator (LASSO) algorithm. Functional annotation of these core biomarkers was performed using consensus clustering and gene set enrichment analysis. We also constructed a protein-protein network and a miRNA-mRNA network using multiple databases, and potential therapeutic agents targeting the core biomarkers were predicted. Finally, we confirmed the expression of the genes in the biomarker panel in both CD and RA using quantitative PCR. RESULTS: A total of five shared core genes, namely C-X-C motif chemokine ligand 10 (CXCL10), C-X-C motif chemokine ligand 9 (CXCL9), aquaporin 9 (AQP9), secreted phosphoprotein 1 (SPP1), and metallothionein 1M (MT1M), were identified as core biomarkers. These biomarkers activate classical pro-inflammatory and immune signaling pathways, influencing immune cell aggregation. Additionally, testosterone was identified as a potential therapeutic agent targeting the biomarkers identified in this study. The expression of genes in the biomarker panel in CD and RA was confirmed through quantitative PCR. CONCLUSION: Our study revealed some core genes shared between CD and RA and established a novel biomarker panel with potential implications for the diagnosis and treatment of these diseases.

OTUB1 suppresses Hippo signaling via modulating YAP protein in gastric cancer.

Gastric cancer is one of the most lethal human malignancies in the world. Although great efforts are put in developing novel therapeutic targets, the effective targeting drugs are still limited. Recent studies reveal the abnormality of Hippo/YAP axis play critical role in the oncogenic process of gastric cancer. It is of great importance to demonstrate the regulation of Hippo signaling activity and YAP protein turnover in gastric cancer. Besides, the phosphorylation cascade on YAP function, which has been thoroughly investigated, the ubiquitination of YAP is also important in Hippo signaling status. Here, We utilized the DUB (Deubiquitinase) siRNA library to identify critical DUB for Hippo signaling. We discovered OTUB1 as a critical factor to facilitate gastric cancer cell stemness and progression, which deubiquitinated and stabilized YAP protein. The clinical data analysis implicated OTUB1 was higher expressed in gastric cancer, which correlated with YAP activity and poor survival. OUTB1 interacted with YAP protein via its OTU domain (Ovarian tumor domain) and deubiquitinated YAP at several lysine sites (K90, K280, K343, K494 and K497), which subsequently inhibited YAP degradation. Our study revealed a novel deubiquitinase of Hippo/YAP axis and one possible therapeutic target for YAP-driven gastric cancer.

The Role of circRNAs in the Diagnosis of Colorectal Cancer: A Meta-Analysis.

Background: A novel category of non-coding circular RNAs (circRNAs) has been found to be dysregulated in colorectal cancer (CRC) and significantly contribute to its progression. However, the feasibility of using circRNA as a diagnostic biomarker for CRC remains to be elucidated. Herein, we aimed to comprehensively collect and analyze evidence regarding the potential application of circRNAs as diagnostic indicators for CRC. Methods: A comprehensive retrieval of relevant studies dating from January, 2015 to December 2020, was carried out in PubMed, Cochrane Library, and Web of Science. Data regarding the diagnostic accuracy of circRNA for CRC, including sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), and area under the curve (AUC), were obtained from the included studies. Quality assessment of diagnostic accuracy studies (QUADAS-2) was used to assess the methodological quality of each study. Statistical analysis was performed using STAT and RevMan software. Results: Eighteen studies, involving a total of 2021 individuals, were included in the present meta-analysis. The specimens examined included tissue, serum, and plasma. The pooled sensitivity, specificity, DOR, PLR, NLR, and AUC, with a 95% confidence interval (CI), of circRNAs in the diagnosis of CRC were 0.78 (0.71-0.83), 0.73 (0.68-0.78), 9.68 (6.76-13.85), 2.92 (2.45-3.50), 0.30 (0.23-0.39), and 0.81 (0.78-0.85), respectively. Subgroup analysis showed that the upregulated circRNAs in the tissue or plasma possessed relatively higher diagnostic values for CRC than the downregulated circRNAs. There was no significant difference between the tissue-derived and non-tissue-derived circRNA subgroups. Conclusion: circRNA may be used as a diagnostic biomarker for CRC because of its relatively high diagnostic accuracy in distinguishing CRC patients from normal controls. Further prospective studies are needed to identify more representative circRNAs as diagnostic markers for CRC.

The E3 Ubiquitin Ligase HOIP inhibits Cancer Cell Apoptosis via modulating PTEN stability.

Chemotherapy is widely used in a variety of solid tumors, such as lung cancer, gastric cancer and breast cancer. The genotoxic drugs, such as cisplatin, suppress cancer progression either by inhibition cell proliferation or facilitating apoptosis. However, the chemotherapy resistance remains an urgent challenge in cancer therapy, especially in advanced stages. Several studies showed that the activation of pro-survival pathways, such as PI3K-AKT, participated in mediating chemotherapy resistance. The insights into the molecular mechanisms for underlying chemotherapy resistance are of great importance to improve cancer patient survival in advanced stages. The HOIP protein belongs to the RING family E3 ubiquitin ligases and modulates several atypical ubiquitination processes in cellular signaling. Previous studies showed that HOIP might be an important effector in modulating cancer cell death under genotoxic drugs. Here, we report that HOIP associates with PTEN and facilitates PTEN degradation in cancer cells. Depletion of HOIP causes cell cycle arrest and apoptosis, which effects could be rescued by PTEN silencing. Besides, the survival data from public available database show that HOIP expression correlates with poor survival in several types of chemotherapy-treated cancer patients. In conclusion, our study establishes a novel mechanism by which HOIP modulates PTEN stability and facilitates chemotherapy resistance in malignancies.

Overexpression of miR-106a enhances oxaliplatin sensitivity of colorectal cancer through regulation of FOXQ1.

Chemotherapy resistance poses a major challenge for the clinical treatment of colorectal cancer, therefore, the aim of the present study was to examine its underlying mechanisms. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to determine the microRNA (miRNA)/mRNA and protein expression levels, respectively. A dual luciferase assay was conducted for verification of the interaction between miR-106a and 3'untranslated region (UTR) of Forkhead box Q1 (FOXQ1). Cell viability was assessed using an MTT assay. In the present study, it was demonstrated that miR-106a is involved in regulating oxaliplatin sensitivity of colorectal cancer. Transfection of miR-106a mimics slightly inhibited colorectal cancer cell growth and sensitized colorectal cancer cells to oxaliplatin exposure. In addition, miR-106a overexpression induced a decrease of FOXQ1 at mRNA and protein levels in colorectal cancer cells. The enhanced expression of miR-106a also increased the expression of Wnt target genes, including vascular endothelial growth factor-A and matrix metallopeptidase 2, which were reported to be regulated by FOXQ1. It was predicted and validated that miR-106a could repress FOXQ1 expression via direct binding to 3'UTR. Elevation of miR-106a and a decrease of FOXQ1 expression levels were detected in tumor tissues from patients with oxaliplatin-sensitive colorectal cancer, compared with patients with oxaliplatin-resistant colorectal cancer. Furthermore, there was a significant association between miR-106a and FOXQ1 mRNA levels. In conclusion, the present study demonstrated that miR-106a increased oxaliplatin sensitivity of colorectal cancer cells through direct repression of FOXQ1 expression.

KIF3B Promotes the Proliferation of Pancreatic Cancer.

Background: Kinesin Family Member 3B (KIF3B) is one of the most ubiquitously expressed KIFs, which is related to numerous physiological responses. KIF3B has also been implicated in carcinogenesis such as in hepatocellular carcinoma cells. However, the expression of KIF3B has not been studied in pancreatic cancer along with its clinical significance. Methods: Immunohistochemical assays were performed to detect the expression levels of KIF3B in the tumor tissues and adjacent non-tumor tissues. Patients were sequentially divided into different expression levels of KIF3B group based on the staining intensity of FKIF3B in tumor tissues. The link between KIF3B expression and clinical characteristics were investigated, and the role of KIF3B on pancreatic cancer cell proliferation was detected by colony formation and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, respectively. And the proliferation related proteins such as Ki67 and proliferating cell nuclear antigen (PCNA) were detected by Western blot. The possible effects of KIF3B on tumor growth were assessed in vivo. Results: KIF3B was highly expressed in human pancreatic cancer tissues. We also found KIF3B was significantly associated to the pTNM stage (*p = 0.018), lymph node metastasis (*p = 0.040) and vascular invasion (*p = 0.034). We reported that increased expression of KIF3B was significantly correlated with poor clinical outcome in our clinical cohort of pancreatic cancer. Furthermore, functional assays revealed that knockdown KIF3B in vitro and in vivo might inhibit cancer cells proliferation by affecting Ki67 and PCNA. Conclusions: Our data suggested that KIF3B was associated with pancreatic cancer malignant progression especially proliferation. Hence, KIF3B might serve as a potential therapy target of pancreatic cancer in clinical treatment.

Silencing of DLEU2 suppresses pancreatic cancer cell proliferation and invasion by upregulating microRNA-455.

Long noncoding RNA (lncRNA) DLEU2 has been shown to be dysregulated in several type of tumor. However, the potential biological roles and molecular mechanisms of DLEU2 in pancreatic cancer (PC) progression are poorly understood. In this study, we found that the DLEU2 level was substantially upregulated in PC tissues and PC cell lines, and significantly associated with poor clinical outcomes in PC patients. Overexpression of DLEU2 significantly induced PC cell proliferation and invasion, whereas knockdown of DLEU2 impaired cell proliferation and invasion in vitro. Furthermore, bioinformatics analysis, luciferase reporter assay, and RNA immunoprecipitation assay revealed that DLEU2 directly bond to microRNA-455 (miR-455) and functioned as an endogenous sponge for miR-455, which could remarkably suppress cell growth and invasion. We also determined that SMAD2 was a direct target of miR-455, and the restoration of SMAD2 rescued cell growth and invasion that were reduced by DLEU2 knockdown or miR-455 overexpression. In addition, low miR-455 expression and high SMAD2 expression was correlated with poor patient survival. These results indicate that DLEU2 is an important promoter of PC development, and targeting the DLEU2/miR-455/SMAD2 pathway could be a promising therapeutic approach in the treatment of PC.

MicroRNA-204 Inhibits the Growth and Motility of Colorectal Cancer Cells by Downregulation of CXCL8.

Among all of the miRNAs, miR-204 has gained considerable attention in the field of cancer research. This study aimed to reveal the detailed functions and the underlying mechanism of miR-204 in colorectal cancer (CRC) cells. The expressions of miR-204 in CRC tumor tissues and cell lines were monitored. Expressions of miR-204 and CXCL8 in Caco-2 and HT-29 cells were altered by transfection, and then cell viability, apoptosis, migration, invasion, EMT-related protein expression, and PI3K/AKT/mTOR pathway protein expression were assessed. We found that miR-204 was expressed at low levels in CRC tumor tissues and cell lines when compared to their normal controls. miR-204 overexpression reduced the viability, migration, and invasion of Caco-2 and HT-29 cells while significantly inducing apoptosis. miR-204 overexpression upregulated E-cadherin expression and downregulated N-cadherin and vimentin expressions. CXCL8 was a target of miR-204, and miR-204 suppression could not increase cell viability, migration, invasion, and EMT procedure when CXCL8 was silenced. Moreover, miR-204 overexpression decreased the phosphorylated levels of PI3K, AKT, and mTOR. The increased phosphorylations of PI3K, AKT, and mTOR, and the upregulation of CXCL8 induced by miR-204 suppression were all abolished by the addition of LY294002 and AZD8055 (inhibitors of PI3K/AKT and mTOR, respectively). To conclude, we demonstrated a tumor-suppressive miRNA in CRC cell lines, miR-204, which is poorly expressed in CRC tissues and cell lines. miR-204 exerted antigrowth, antimigration, anti-invasion, and anti-EMT activities, which might be via deactivating the PI3K/AKT/mTOR pathway and repressing CXCL8 expression.

miR-522-3p Promotes Tumorigenesis in Human Colorectal Cancer via Targeting Bloom Syndrome Protein.

miR-522-3p is known to degrade bloom syndrome protein (BLM) and enhance expression of other proto-oncogenes, leading to tumorigenesis. This study aimed to investigate the molecular mechanisms of miR-522-3p in human colorectal cancer (CRC) cells. Expressions of miR-522-3p in CRC and adjacent tissues, as well as in normal human colon epithelial cell line (FHC) and five CRC cell lines, were detected. Human CRC cell lines, HCT-116 and HT29, were transfected with miR-522-3p mimic, inhibitor, or scrambled controls. Then cell viability, apoptosis, cell cycle progression, and the expressions of c-myc, cyclin E, CDK2, and BLM were assessed. It was found that miR-522-3p was highly expressed in CRC tissues when compared to adjacent nontumor tissues and was highly expressed in CRC cell lines when compared to FHC cells. miR-522-3p overexpression promoted cell viability, reduced apoptotic cell rate, arrested more cells in the S phase, and upregulated c-myc, cyclin E, and CDK2 expression. BLM was a target gene of miR-522-3p, and miR-522-3p suppression did not exert antiproliferative and proapoptotic activities when BLM was silenced. These findings demonstrate that miR-522-3p upregulation negatively regulates the expression of BLM, with upregulation of c-myc, CDK2, and cyclin E, and thereby promoting the proliferation of human CRC cells.

The diagnostic and prognostic value of CHFR hypermethylation in colorectal cancer, a meta-analysis and literature review.

The Checkpoint with Forkhead-associated and Ring finger domains (CHFR) is a mitotic checkpoint and tumor-suppressor gene, its loss contributes tumorigenesis of epithelial cancers including colorectal carcinoma (CRC). The diagnostic and prognostic value of CHFR promoter hypermethylation in CRC remains unclear. This study aimed to conduct a meta-analysis and literature review and investigate clinicopathological significance of CHFR promoter hypermethylation in CRC. The following online database were used: PubMed, EMBASE, and Web of Science up to March 2017. Odds Ratios (OR) and Hazard Ratios (HR) with 95% corresponding confidence intervals (CIs) were calculated. A total of seven relevant articles were available for meta-analysis which included 966 patients. The frequency of CHFR promoter hypermethylation significantly increased in CRC compared to normal colorectal mucosa tissue, pooled OR was 8.35, p < 0.00001. CHFR promoter hypermethylation was not significantly correlated to stage, OR was 1.16, p = 0.63. However, CHFR promoter hypermethylation was more frequently observed in CRC with positive lymph nodes metastasis than CRC with negative lymph nodes metastasis, OR was 0.46, p = 0.03. Additionally CHFR promoter hypermethylation was significantly related to poor overall survival in patients with CRC, HR was 0.62, p = 0.008. Based on these results, tumor CHFR promoter hypermethylation is not only a diagnostic biomarker for CRC, but also a prognostic marker. CHFR promoter hypermethylation is significantly associated with worse overall survival in patients with CRC. Our data suggested that CHFR could be a potential drug target for development of demethylation treatment for patients with CRC.

APC hypermethylation for early diagnosis of colorectal cancer: a meta-analysis and literature review.

Adenomatous polyposis coli (APC) promoter hypermethylation has been frequently observed in colorectal cancer (CRC). The association between APC promoter methylation and clinicopathological significance in CRC is under investigation. We performed a meta-analysis to quantitatively evaluate the significance of APC methylation in CRC. The study included a total of 24 articles and 2025 CRC patients. The frequency of APC promoter hypermethylation was significantly higher in colorectal adenoma than in normal colorectal tissue, OR was 5.76, 95% CI, 2.45-13.56; p<0.0001, I2=0%. APC promoter more frequently hypermethylated in CRC stage I compared to normal colorectal tissue, OR was 13.42, 95% CI, 3.66-49.20; p<0.0001, I2=31%. The risk of incidence of CRC was significantly correlated to APC promoter hypermethylation, pooled OR was 9.80, 95%CI, 6.07-15.81; p<0.00001, I2=43%. APC methylation was not associated with grade, stage of CRC as well as tumor location, patients' gender, and smoking behavior. The results indicate that APC promoter hypermethylation is an early event in carcinogenesis of CRC, could be a valuable diagnostic marker for early-stage CRC. APC methylation is not significantly associated with overall survival in patients with CRC. APC is a potential drug target for development of personalized treatment.

miR-203 promotes proliferation, migration and invasion by degrading SIK1 in pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDA) is among the most lethal human cancers and it is insensitive to many chemotherapeutic drugs. The molecular basis of pancreatic cancer remains to be elucidated. Investigations into the molecular mechanism involved in the development and progression as well as drug resistance of the disease may be useful to understand the pathogenesis and progression of the disease and offer new targets for effective therapies. In the present study, we showed that salt-inducible kinase 1 (SIK1) was downregulated and loss of SIK1 was associated with gemcitabine resistance in pancreatic cancer. In pancreatic cancer cells, SIK1 inhibited proliferation, migration and invasion. An analysis of potential microRNA target sites was performed using the prediction algorithms, miRanda, TargetScan and PicTar. The three algorithms predicted that miR-203 is capable of targeting 3'UTR of SIK1. Subsequent experiments confirmed the prediction. In addition, the results showed that miR-203 promotes proliferation, migration and invasion in pancreatic cancer cells, whereas the restoration of SIK1 abrogated the regulation of pre-miR­203-mediated proliferation, migration and invasion.

Accumulation of foxp3+ T regulatory cells in draining lymph nodes correlates with disease progression and immune suppression in colorectal cancer patients.

PURPOSE: To assess the relation of Foxp3(+) regulatory T cells (Treg) in tumor draining lymph nodes (TDLNs) with tumor progression and immune suppression in colorectal cancer (CRC). EXPERIMENTAL DESIGN: Flow cytometry was used to analyze the densities of Tregs in lymphocytes of TDLNs, peripheral blood, and tumors from 34 patients with CRC. The frequency of Tregs was compared and evaluated for the association with disease stage. The effect of Tregs on the function of CD8(+) T cells was investigated by IFN-gamma production. RESULTS: The density of Foxp3(+) Tregs in TDLNs was dramatically higher than that in peripheral blood lymphocytes, but significantly lower than that in tumor-infiltrating lymphocytes. Importantly, the frequency of Foxp3(+) Tregs in TDLNs, rather than that in tumors and peripheral blood, was positively correlated with disease stage. In addition, the functions of CD8(+) T cells were impaired in TDLNs compared with peripheral blood lymphocytes and were restored after Treg depletion. CONCLUSIONS: Foxp3(+) Tregs in TDLNs are more correlated with disease progression and potentially influence CD8(+) T-cell functions. This study suggests that the frequency of Tregs in TDLNs may provide a valuable prognostic tool in the treatment of CRC.

Surgical management for carcinoma of the gallbladder: a single-institution experience in 25 years.

BACKGROUND: Gallbladder carcinoma is rare and associated with dismal outcomes. Radical surgery is the only curative treatment, and options for adjuvant therapy remain limited. This study aimed to determine the factors influencing outcome of treatment in patients with gallbladder carcinoma, and to identify the patients who might benefit from radical surgery and adjuvant therapy. METHODS: Medical records and follow-up histories of 150 patients with gallbladder carcinoma who had undergone surgery between April 1980 and December 2005 were retrospectively reviewed. The factors predictive for the survival of the patients were identified using multivariate analysis. RESULTS: Surgery for gallbladder cancer was associated with an overall 5-year survival rate of 26.2%. After curative resection (40% of the patients), the 5-year survival rate was 60.3%. The patients who underwent R0 resection had a significantly longer median survival (97.3 months) than those who had R1/R2 resection (8.3 months) or only laparotomy (3.7 months) (P < 0.0001). Univariate analysis showed that resectability, American Joint Committee on Cancer staging, tumor grade, adjuvant therapy, jaundice at presentation, depth of tumor invasion, lymph node involvement, distant metastasis, and carcinoembryonic antigen level were statistically significant predictors for survival. Multivariate analysis revealed American Joint Committee on Cancer staging and resectability were independent prognostic factors for survival. The patients who underwent noncurative resection might benefit from adjuvant therapy (median survival, 12.4 months vs 7.2 months, P = 0.006). CONCLUSIONS: Favorable survival rate can be achieved after curative resection, even for selected patients with advanced disease. Adjuvant therapy may improve the survival of patients with gallbladder carcinoma.

[Simultaneous liver and colorectal resection for synchronous colorectal liver metastasis].

OBJECTIVE: To investigate the safety and long term outcome of simultaneous liver and colorectal resection for synchronous colorectal liver metastasis. METHODS: Forty-three synchronous colorectal liver metastasis patients who received simultaneous colectomy and hepatectomy between May 1981 and November 2005 were analyzed retrospectively. RESULTS: The group included 21 male patients and 22 female patients, with the median age of 52 years. The overall median operative time was 180 minutes, 30 cases received blood transfusion, and the median volume was 800 ml. The median hospital stay was 15 days. The morbidity and mortality was 18.6% and 2.3%, respectively. The overall median survival time was 25 months, 5-year survival rate was 19.1%. The survival of patients underwent R0 resection were substantially better (median survival time 48 months, 5-year survival rate 33.8%) than that of the patients who did not undergo R0 resection (20 months, 7.6%) (P = 0.002). CONCLUSIONS: Simultaneous liver and colorectal resection is safe and effective for synchronous colorectal liver metastasis. Furthermore, simultaneous R0 resection should be the optimal surgery for the resectable cases.

[Outcome of surgical therapy for liver metastasis of colorectal cancer: analysis of 75 cases].

OBJECTIVE: To explore the strategy to improve the long term survival of liver metastasis of colorectal cancer after surgical treatment. METHODS: The clinical data of 75 patients with liver metastasis of colorectal cancer, 43 males and 32 females, aged 51.4, who received hepatectomy between January 1981 and November 2005, were analyzed. RESULTS: The primary tumor site was colon in 39 cases, and rectum in 36 cases. Liver metastasis was synchronous in 59 patients, and metachronous in 16 patients. 45 patients received simultaneous liver and colorectal resection, 29 patients received metachronous resection, and 1 patient did not receive primary rectal cancer resection. The operative complication rate and the mortality were 16% (12/75) and 1.33% (1/75) respectively. The overall 1- 3-, and 5-year survival rates were 86.7%, 35.5%, and 22.2% respectively, and the median survival time was 25 months. There were residual tumors in 35 patients. The 1-, 3-, and 5-year survival rates of the residual tumor group were 80.6%, 5.4%, and 5.4% respectively, all significantly lower than those of the radical resection group (91.6%, 58.1%, and 34.9% respectively, and the median survival time of the residual tumor group was 18 months, significantly shorter than that of the radical resection group (38 months) (all P = 0.000). CONCLUSION: Surgical resection of liver metastasis of colorectal cancer significantly prolongs the survival time, and resection of all liver deposits and the extrahepatic disease is the most important factor influencing survival.

[Clinicopathological features and surgical treatment of familial adenomatous polyposis].

OBJECTIVE: To investigate the clinicopathological features and surgical treatment of familial adenomatous polyposis (FAP). METHODS: The clinical data of 51 patients with FAP, 29 males and 22 females, aged 36.6 (17-67), hospitalized Sep 1980-Dec 2005 were analyzed. RESULTS: Twenty-nine patients had family history of FAP and 7 of them were discovered during screened because of family history. The main clinical manifestations included rectal bleeding or bloody stool, diarrhea, abdominal pain, and general discomfort in the abdomen. Synchronous malignant changes were found in 28 patients while hospitalized. The incidence of colorectal cancer was 54.9%. The average age of those with colorectal cancer was 40.5 and the age of those without colorectal cancer was 31.9. 81.8% of the colorectal cancers occurred on the rectum and sigmoid colon. Different types of proctocolectomy were performed on 50 patients, and one patient received only pelvic radiotherapy. The follow-up rate was 74.5%. Only 27.5% of the patients received regular surveillance. Colorectal cancer was found in 8 patients after the first operation. So far 17 patients died. CONCLUSIONS: There were no specific clinical manifestations for FAP. Colonoscopy is the most important diagnostic procedure. Surgical treatment should be given as soon as possible. The specific operation type should be based on the individual aspects of patients. Life-time surveillance and follow-up are very important.