ABSTRACT
BACKGROUND: Monopolar spindle-binding protein 3B (MOB3B) functions as a signal transducer and altered MOB3B expression is associated with the development of human cancers. AIM: To investigate the role of MOB3B in colorectal cancer (CRC). METHODS: This study collected 102 CRC tissue samples for immunohistochemical detection of MOB3B expression for association with CRC prognosis. After overexpression and knockdown of MOB3B expression were induced in CRC cell lines, changes in cell viability, migration, invasion, and gene expression were assayed. Tumor cell autophagy was detected using transmission electron microscopy, while nude mouse xenograft experiments were performed to confirm the in-vitro results. RESULTS: MOB3B expression was reduced in CRC vs normal tissues and loss of MOB3B expression was associated with poor CRC prognosis. Overexpression of MOB3B protein in vitro attenuated the cell viability as well as the migration and invasion capacities of CRC cells, whereas knockdown of MOB3B expression had the opposite effects in CRC cells. At the molecular level, microtubule-associated protein light chain 3 II/I expression was elevated, whereas the expression of matrix metalloproteinase (MMP)2, MMP9, sequestosome 1, and phosphorylated mechanistic target of rapamycin kinase (mTOR) was downregulated in MOB3B-overexpressing RKO cells. In contrast, the opposite results were observed in tumor cells with MOB3B knockdown. The nude mouse data confirmed these in-vitro findings, i.e., MOB3B expression suppressed CRC cell xenograft growth, whereas knockdown of MOB3B expression promoted the growth of CRC cell xenografts. CONCLUSION: Loss of MOB3B expression promotes CRC development and malignant behaviors, suggesting a potential tumor suppressive role of MOB3B in CRC by inhibition of mTOR/autophagy signaling.
Subject(s)
Autophagy , Cell Movement , Colorectal Neoplasms , Neoplasm Invasiveness , Signal Transduction , TOR Serine-Threonine Kinases , Aged , Animals , Female , Humans , Male , Mice , Middle Aged , Adaptor Proteins, Signal Transducing/metabolism , Adaptor Proteins, Signal Transducing/genetics , Cell Line, Tumor , Cell Survival , Colorectal Neoplasms/pathology , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/genetics , Down-Regulation , Gene Expression Regulation, Neoplastic , Gene Knockdown Techniques , Mice, Inbred BALB C , Mice, Nude , Prognosis , TOR Serine-Threonine Kinases/metabolismABSTRACT
Colorectal cancer (CRC) is a common malignancy that has the second highest incidence and mortality rate. Although there are many personalized treatment options for CRC, the therapeutic effects are ultimately limited by drug resistance. Studies have aimed to block the initiation and progression of CRC by inducing cell death to overcome this obstacle. Substantial evidence has indicated that both autophagy and ferroptosis play important regulatory roles in CRC. Autophagy, a lysosome-dependent process by which cellular proteins and organelles are degraded, is the basic mechanism for maintaining cell homeostasis. The duality and complexity of autophagy in cancer therapy is a hot topic of discussion. Ferroptosis, a regulated cell death pathway, is associated with iron accumulation-induced lipid peroxidation. The activation of ferroptosis can suppress CRC proliferation, invasion and drug resistance. Furthermore, recent studies have suggested an interaction between autophagy and ferroptosis. Autophagy can selectively degrade certain cellular contents to provide raw materials for ferroptosis, ultimately achieving antitumor and anti-drug resistance. Therefore, exploring the interaction between autophagy and ferroptosis could reveal novel ideas for the treatment of CRC. In this review, we describe the mechanisms of autophagy and ferroptosis, focusing on their roles in CRC and the crosstalk between them.
ABSTRACT
BACKGROUND AND AIM: The biological functions and clinical implications of lysophosphatidylcholine acyltransferase 1 (LPCAT1) remain unclarified in gastric cancer (GC). The aim of the current study was to explore the possible clinicopathological significance of LPCAT1 and its perspective mechanism in GC tissues. MATERIALS AND METHODS: The protein expression and mRNA levels of LPCAT1 were detected from in-house immunohistochemistry and public high-throughput RNA arrays and RNA sequencing. To have a comprehensive understanding of the clinical value of LPCAT1 in GC, all enrolled data were integrated to calculate the expression difference and standard mean difference (SMD). The biological mechanism of LPCAT1 in GC was confirmed by computational biology and in vitro experiments. Migration and invasion assays were also conducted to confirm the effect of LPCAT1 in GC. RESULTS: Both protein and mRNA expression levels of LPCAT1 in GC were remarkably higher than those in noncancerous controls. Comprehensively, the SMD of LPCAT1 mRNA was 1.11 (95% CI = 0.86-1.36) in GC, and the summarized AUC was 0.85 based on 15 datasets containing 1727 cases of GC and 940 cases of non-GC controls. Moreover, LPCAT1 could accelerate the invasion and migration of GC by boosting the neutrophil degranulation pathway and disturbing the immune microenvironment. CONCLUSION: An increased level of LPCAT1 may promote the progression of GC.
Subject(s)
Stomach Neoplasms , Humans , Stomach Neoplasms/genetics , 1-Acylglycerophosphocholine O-Acyltransferase/genetics , 1-Acylglycerophosphocholine O-Acyltransferase/metabolism , Cell Proliferation , Acyltransferases , Computational Biology , RNA, Messenger/genetics , Tumor MicroenvironmentABSTRACT
Chinese herb Radix sophorae tonkinensis extract oxymatrine shows anticancer effects. This study evaluated the role of oxymatrine in colorectal cancer (CRC) and the underlying molecular events in vitro and in vivo. CRC cells were treated with different doses of oxymatrine to assess cell viability, reactive oxygen species production, gene expression, and gene alterations. Meanwhile, mouse xenograft and liver metastasis models were used to assess the effects of oxymatrine using histology examination, transmission electron microscopy, and Western blot, respectively. Our results showed that oxymatrine treatment triggered CRC cell mitophagy to inhibit CRC cell growth, migration, invasion, and metastasis in vitro and in vivo. At the gene level, oxymatrine inhibited LRPPRC to promote Parkin translocation into the mitochondria and reduce the mitophagy-activated NLRP3 inflammasome. Thus, oxymatrine had an anticancer activity through LRPPRC inhibition, mitophagy induction, and NLRP3 inflammasome suppression in the CRC cell xenograft and liver metastasis models. In conclusion, the study demonstrates the oxymatrine anti- CRC activity through its unique role in regulating CRC cell mitophagy and NLRP3 inflammasome levels in vitro and in vivo.
Subject(s)
Alkaloids , Colorectal Neoplasms , Liver Neoplasms , Humans , Mice , Animals , Inflammasomes/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Mitophagy/physiology , Alkaloids/pharmacology , Colorectal Neoplasms/drug therapy , Liver Neoplasms/drug therapyABSTRACT
BACKGROUND: Small intestinal cavernous hemangioma is a rare disease, especially in the ileum. It is difficult to accurately diagnose due to its hidden location and nonspecific clinical symptoms. Here, we reported a case of ileal cavernous hemangioma with chronic hemorrhage in a 20-year-old man and review the literature to gain a better understanding of this disease. CASE SUMMARY: The patient complained of intermittent melena and hematochezia for > 3 mo. The lowest hemoglobin level revealed by laboratory testing was 3.4 g/dL (normal range: 12-16 g/dL). However, the gastroscopy, colonoscopy and peroral double-balloon enteroscopy (DBE) showed no signs of bleeding. The transanal DBE detected a lesion at about 340 cm proximal to the ileocecal valve. Thus, we performed an exploratory laparoscopy and the lesion was resected. After the operation, the patient had no melena. Finally, the pathological examination identified the neoplasm as an ileal cavernous hemangioma, thereby resulting in gastrointestinal hemorrhage. CONCLUSION: This report might improve the diagnosis and treatment of ileal cavernous hemangioma.
ABSTRACT
Invasion and metastasis are the main causes of colorectal cancer (CRC)-related death. Accumulating evidence suggested that sphingosine kinase 1 (SphK1) promoted the metastasis of CRC and autophagy played an important role in SphK1 promoting the metastasis of malignancy. However, the mechanism by which SphK1-driven autophagy promotes invasion and metastasis in CRC remains to be clarified. In the present study, immunohistochemical detection showed the expression of SphK1 and paxillin was higher in human CRC tissues than those of normal colorectal mucosal tissues, they were both associated with TNM staging, lymphatic, and distance metastasis. In addition, study of in situ tumor transplantation model in nude mice showed that the suppression of SphK1 inhibited the growth of colonic orthotopic implantation tumors and the expression of paxillin, p-paxillin, LC3 in the tumor. So, SphK1 may promote CRC metastasis via inducing the expression of paxillin expression and its phosphorylation, in vivo. Furthermore, results of CCK8 assay, transwell and wound healing assays showed that SphK1 promoted the viability, invasion, and metastasis of CRC cells. Transmission electron microscopy detection showed that SphK1 is the key factor in autophagy induction in CRC cells. Moreover, western blot examination indicated that the expression of LC3â ¡/â , paxillin, p-paxillin, MMP-2, and vimentin was enhanced in SphK1-overexpressed CRC cells and suppressed in SphK1 knockdown CRC cells, meanwhile, the expression of E-cadherin was suppressed in SphK1-overexpressed CRC cells and enhanced in SphK1 knockdown CRC cells. Suppression of autophagy by 3MA reversed the expression of paxillin and its phosphorylation in SphK1-overexpressed CRC cells, indicated that SphK1-driven autophagy induced the expression of paxillin and its phosphorylation in CRC cells. Together, these findings reveal that SphK1-driven autophagy may promote the invasion and metastasis of CRC via promoting the expression of focal adhesion paxillin and its phosphorylation.
Subject(s)
Autophagy/genetics , Focal Adhesions/metabolism , Phosphotransferases (Alcohol Group Acceptor)/metabolism , Adult , Aged , Aged, 80 and over , Animals , Colorectal Neoplasms/genetics , Female , Humans , Male , Mice , Mice, Nude , Middle Aged , Neoplasm MetastasisABSTRACT
Tricellulin is a tightjunction transmembrane protein that regulates cellcell interactions. Altered tricellulin expression could promote tumor cell invasions and metastasis in human cancers. The present study assessed tricellulin expression in colorectal cancer tissues for any association with clinicopathological features of colorectal cancer patients and then investigated the underlying molecular events using quantitative proteomic analysis and in vitro experiments. Tissue samples from 98 colorectal cancer patients and 15 volunteers were collected for immunohistochemistry. Colorectal cell lines were used to overexpress or knockdown tricellulin expression in various assays. The data revealed that upregulated tricellulin expression was associated with lymph node and distant metastases and poor prognosis, while tricellulin overexpression promoted colorectal cancer cell migration and invasion in vitro. In contrast, tricellulin knockdown had positive effects on the tumor cells. Furthermore, TMTLCMS/MS and bioinformatics analyses revealed that tricellulin was involved in EMT and reduction of apoptosis through the NFκB signaling pathway. These findings highlight for the first time the significance of tricellulin in colorectal cancer development and progression. Further study may validate tricellulin as a novel biomarker and target for colorectal cancer.
Subject(s)
Adenocarcinoma/secondary , Biomarkers, Tumor/metabolism , Carcinogenesis/pathology , Colorectal Neoplasms/pathology , MARVEL Domain Containing 2 Protein/metabolism , Adenocarcinoma/diagnosis , Adenocarcinoma/mortality , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Biomarkers, Tumor/genetics , Case-Control Studies , Cell Line, Tumor , Cell Movement , Cell Proliferation , Colorectal Neoplasms/diagnosis , Colorectal Neoplasms/mortality , Computational Biology , Disease Progression , Epithelial-Mesenchymal Transition , Female , Gene Knockdown Techniques , Healthy Volunteers , Humans , Immunohistochemistry , MARVEL Domain Containing 2 Protein/analysis , MARVEL Domain Containing 2 Protein/genetics , Male , Middle Aged , NF-kappa B/metabolism , Prognosis , Signal TransductionABSTRACT
OBJECTIVES: This study aimed to investigate hub genes and their prognostic value in colon cancer via bioinformatics analysis. METHODS: Differentially expressed genes (DEGs) of expression profiles (GSE33113, GSE20916, and GSE37364) obtained from Gene Expression Omnibus (GEO) were identified using the GEO2R tool and Venn diagram software. Function and pathway enrichment analyses were performed, and a protein-protein interaction (PPI) network was constructed. Hub genes were verified based on The Cancer Genome Atlas (TCGA) and Human Protein Atlas (HPA) databases. RESULTS: We identified 207 DEGs, 62 upregulated and 145 downregulated genes, enriched in Gene Ontology terms "organic anion transport," "extracellular matrix," and "receptor ligand activity", and in the Kyoto Encyclopedia of Genes and Genomes pathway "cytokine-cytokine receptor interaction." The PPI network was constructed and nine hub genes were selected by survival analysis and expression validation. We verified these genes in the TCGA database and selected three potential predictors (ZG16, TIMP1, and BGN) that met the independent predictive criteria. TIMP1 and BGN were upregulated in patients with a high cancer risk, whereas ZG16 was downregulated. The immunostaining results from HPA supported these findings. CONCLUSION: This study indicates that these hub genes may be promising prognostic indicators or therapeutic targets for colon cancer.
Subject(s)
Colonic Neoplasms , Computational Biology , Colonic Neoplasms/genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , HumansABSTRACT
BACKGROUND: Many epidemiological studies have investigated the risk factors for clonorchiasis, but endoscopic findings of this disease in endoscopic retrograde cholangiopancreatography (ERCP) have not been well characterized. In this study, we evaluated clonorchiasis in ERCP in patients with biliary and pancreatic diseases. METHODS: This was a retrospective two-center study in hospitalized patients who received ERCP between January 2012 and October 2018. All patients were divided into clonorchiasis and nonclonorchiasis groups. Data were analyzed using univariate analysis and multivariate analyses. RESULTS: A total of 1119 patients were included, and clonorchiasis was diagnosed in 19.2% patients. Detection of Clonorchis sinensis eggs in bile samples was higher than that in fecal samples (85.9% vs. 58.7%; P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249, P = 0.001). In multivariate analysis, male patients (95% confidence interval (CI): 1.945-4.249. CONCLUSIONS: The detection of C. sinensis eggs was significantly higher in bile than in fecal samples; thus, bile samples represent a preferable sample for the diagnosis of clonorchiasis in patients with biliary obstruction. We found that male, age ≤ 60 years old, and CBD diameter < 12 mm were independent risk factors for clonorchiasis, while papilla fistula was a protective factor.C. sinensis eggs was significantly higher in bile than in fecal samples; thus, bile samples represent a preferable sample for the diagnosis of clonorchiasis in patients with biliary obstruction. We found that male, age ≤ 60 years old, and CBD diameter < 12 mm were independent risk factors for clonorchiasis, while papilla fistula was a protective factor.
Subject(s)
Bile/parasitology , Clonorchiasis/diagnosis , Clonorchiasis/etiology , Pancreatic Diseases/parasitology , Adult , Aged , Aged, 80 and over , Animals , Cholangiopancreatography, Endoscopic Retrograde/methods , Clonorchis sinensis/parasitology , Duodenum/parasitology , Feces/parasitology , Female , Humans , Male , Middle Aged , Retrospective Studies , Risk Factors , Young AdultABSTRACT
The present study investigated the sensitization of 5-fluorouracil (5-FU)-resistant colon cancer cells in vitro, using oxymatrine, a Chinese herb, and a quinolizidine alkaloid compound extracted from the root of Sophora flavescens. The HCT-8 colon cancer cell line and its 5-FU-resistant subline HCT-8/5-FU were treated with 5-FU and oxymatrine, alone or in combination, at various doses. The cells were subsequently assessed for changes in cell viability, apoptosis and morphology and analyzed by fluorescence microscopy and western blotting. The data demonstrated that HCT-8/5-FU markedly increased the dose of 5-FU required for the suppression of tumor cell viability (78.77±1.90 µg/ml vs. 9.20±0.96 µg/ml in parental HCT-8 cells), whereas HCT-8/5-FU induced the tumor cell epithelial-mesenchymal transition (EMT). By contrast, oxymatrine alone and in combination with 5-FU altered HCT-8/5-FU cell morphology, apoptosis and EMT phenotypes. The combination of oxymatrine and 5-FU reduced the protein expression of snail family transcriptional repressor 2 and vimentin, phosphorylated p65 and induced the expression of E-cadherin, by inhibiting the nuclear factor κB (NF-κB) signaling pathway. In conclusion, the data from the present study demonstrated that EMT was associated with 5-FU chemoresistance in HCT-8/5-FU colon cancer cells, and that oxymatrine treatment was able to reverse such resistance. Oxymatrine may regulate tumor cell EMT and inactivate the NF-κB signaling pathway, and may therefore serve as a potential therapeutic drug to reverse 5-FU resistance in colon cancer cells.
ABSTRACT
BACKGROUND: The bile infection may already exist before the administration of an interventional procedure, despite no clinical manifestations of cholangitis detected. Blood cultures remained negative even in more than half of the febrile cases with cholangitis. Risk factors associated with bacterial growth in bile before the intervention are not well defined. To establish the bacterial profiles isolated from the bile samples and to identify risk factors for bacterial colonization in the bile system. METHODS: Individuals who underwent endoscopic retrograde cholangiopancreatography (ERCP) interventions were enrolled. Bile samples were aspirated and were immediately transferred into a sterile tube for storage. RESULTS: Positive bile cultures were detected in 363 (38.0%) of 956 patients, including 322 benign diseases and 41 malignances. Of 363 positive cases, 351 (96.7%) were monoinfection and 12 (3.3%) multi-infection. Escherichia coli were the most common Gram-negative bacteria (210, 56.0%), followed by Klebsiella pneumoniae (45, 12.0%). Enterococcus faecalis represented the most common Gram-positive microorganism (19, 5.07%), while Candida albicans (11, 2.93%) were the dominant fungi. Klebsiella pneumoniae were more frequently detected in malignant diseases (P = 0.046). Age, previous ERCP history or OLT history, and CBD diameter were independent risk factors for positive cultures (P < 0.05) while preoperative jaundice drug therapy was the protective factor for bile infection (P < 0.05). CONCLUSION: Monomicrobial infection was dominant among all infections, and Klebsiella pneumoniae strains were more frequently isolated from patients with malignant diseases. To effectively manage patients who are at a high risk for bile infection, a detailed diagnosis and treatment plan for each case should be prepared.
ABSTRACT
It was demonstrated that Sphingosine kinase 1 (SphK1) promotes tumor progression and confers the malignancy phenotype of colorectal cancer by activating the focal adhesion kinase (FAK) pathway. However, further clarification is required to determine if SphK1 promotes the metastasis of colorectal cancer by inducing epithelialmesenchymal transition (EMT), and its mechanisms have not been fully elucidated. Immunohistochemistry staining was used to detect protein expression in normal colonic mucosa tissues and colorectal cancer tissues. Cells were transfected to overexpress SphK1, downregulate SphK1 or downregulate FAK. An MTT assay was used to detect the drug toxicity to cells. Transwell and wound healing assays were used to detect cell migration ability. Reverse transcriptionpolymerase chain reaction and western blot analysis were used to detect the expression of mRNA and protein, respectively. Scanning electron microscopy was used to observe the microvilli and pseudopodia of the cells. The analysis of protein expression in 114 human colorectal cancer tissues demonstrated that the expressions of SphK1, FAK, phosphorylated (p)FAK, Ecadherin and vimentin were associated with the metastasis of colorectal cancer. Furthermore, the patients with colorectal cancer with SphK1positive cancer demonstrated poorer prognosis compared with SphK1negative cancer. FAK knockdown and SphK1 knockdown of human colon cancer RKO cells inhibited the EMT and migrational potency, along with the expression of pFAK, pprotein kinase B (AKT) and matrix metalloproteinase (MMP)2/9. In contrast, SphK1 overexpression promoted EMT, migrational potency, and the expression of pFAK, pAKT and MMP2/9 in HT29 cells. Additionally, the EMT and migrational potency of SphK1overexpressing HT29 cells was suppressed by a FAK inhibitor, and the expression of pFAK, pAKT and MMP2/9 was suppressed by blocking the FAK pathway. In conclusion, SphK1 promoted the migration and metastasis of colon cancer by inducing EMT mediated by the FAK/AKT/MMPs axis.
Subject(s)
Colorectal Neoplasms/metabolism , Phosphotransferases (Alcohol Group Acceptor)/genetics , Phosphotransferases (Alcohol Group Acceptor)/metabolism , Signal Transduction , Adult , Aged , Aged, 80 and over , Cell Line, Tumor , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Epithelial-Mesenchymal Transition , Female , Focal Adhesion Protein-Tyrosine Kinases/metabolism , HT29 Cells , Humans , Male , Matrix Metalloproteinases/metabolism , Middle Aged , Neoplasm Metastasis , Neoplasm Staging , Proto-Oncogene Proteins c-akt/metabolism , Survival AnalysisABSTRACT
Systematic chemotherapy is indispensable for gastric cancer patients with advanced stage disease, but the occurrence of chemoresistance drastically limits treatment effectiveness. There is a tremendous need for identifying the underlying mechanism of chemoresistance. NIK and IKKßbinding protein (NIBP) (also known as TRAPPC9, trafficking protein particle complex 9) is a regulator of the cytokineinduced NFκB signaling pathway which has been proven to play pivotal roles in the progression of various malignancies. Nevertheless, it is still ambiguous whether NIBP is involved in the chemoresistance of gastric cancer. The aim of the present study was to investigate the effect of NIBP on chemotherapy resistance of gastric cancer (GC) and to research the mechanisms of Ginkgo biloba extract 761 (EGb 761®) on reversing chemoresistence which has been confirmed in our previous study. In the present study, the results of immumohistochemisty revealed that the positive staining rates of NIBP, NFκB p65 and NFκB pp65 in gastric cancer tissues were obviously higher than those in normal tissues. Furthermore, a close correlation was found to exist between the expression of NIBP and NFκB p65 (pp65) in gastric cancer tissues. Moreover, the overexpression of NIBP was closely related to tumor differentiation, depth of invasion, clinical stage and lymphatic metastasis in gastric cancer. Western blot analysis, realtime PCR, MTT assay and flow cytometric analysis were performed and the results demonstrated that compared with the gastric cancer SGC7901 cells, the expression of NIBP, NFκB p65, NFκB pp65 and mesenchymal marker vimentin were significantly increased in gastric cancer multidrugresistant SGC7901/CDDP cells, and the epithelial cell marker ZO1 was significantly decreased. Meanwhile, it was found that SGC7901/CDDP cells were accompanied by spindlelike mesenchymal appearance and upregulation of stem cell marker CD133 which has been verified to be an upstream regulatory gene of epithelialmesenchymal transition (EMT). Further research confirmed that downregulation of NIBP by Ginkgo biloba extract (EGb) 761 EGb 761 suppressed the cisdiamminedichloroplatinum(II) (CDDP)induced NFκB signaling pathway, EMT and the expression of CD133 in SGC7901 and SGC7901/CDDP cells. Altogether, these data indicate that the NIBPregulated NFκB signaling pathway plays a pivotal role in the chemoresistance of gastric cancer by promoting CD133induced EMT.
Subject(s)
Carrier Proteins/metabolism , Drug Resistance, Neoplasm , Epithelial-Mesenchymal Transition , Signal Transduction , Stomach Neoplasms/metabolism , Up-Regulation , Adult , Aged , Carrier Proteins/genetics , Cell Line, Tumor , Cell Survival/drug effects , Drug Resistance, Neoplasm/drug effects , Epithelial-Mesenchymal Transition/drug effects , Female , Ginkgo biloba , Humans , Intercellular Signaling Peptides and Proteins , Male , Middle Aged , NF-kappa B/metabolism , Plant Extracts/pharmacology , Signal Transduction/drug effects , Stomach Neoplasms/genetics , Up-Regulation/drug effectsABSTRACT
OBJECTIVE: To determine the presence of vasculogenic mimicry (VM) and expression of Sphingosine kinase 1 (SphK1) and Connexin43 (Cx43) in colorectal cancer (CRC) tissues, and to identify their inter-relationships and associations with multiple pathologic parameters. METHODS: Ninety-two CRC specimens and normal pericarcinoma tissues were analyzed for expression of SphK1 and Cx43 using immunohistochemistry, and for identification of VM using CD34-periodic acid-Schiff dual staining. RESULTS: The positive rate of SphK1 expression was greater in CRC cells than pericarcinoma cells (85.87% vs. 33.70%, P < 0.05). In contrast, the positive rate of Cx43 expression was greater in pericarcinoma cells than in CRC cells (58.70% vs. 92.39%, P < 0.05). Analysis of CRC tissues indicated that expression of SphK1 was associated with poor differentiation, advanced tumor stage, lymph node metastasis, and the presence of VM (P < 0.05 for each comparison). Expression of Cx43 was associated with high differentiation and the presence of VM (P < 0.05 for each comparison). Patient sex, age, tumor size, depth of invasion, and distant metastasis were unrelated to the expression of either protein. There was a significant correlation between the expression of SphK1 and Cx43 (P < 0.05). Analysis of overall patient survival indicated that SphK1 positivity and the presence of VM were significantly associated with poor survival, but Cx43 positivity had no relationship with survival. CONCLUSION: SphK1 protein expression was significantly greater in CRC tissues than pericarcinoma tissues, suggesting this protein may be associated with the pathogenesis of CRC. In addition, the significant correlation between expression of SphK1 and Cx43 in CRC tissues suggests their interaction may impact the pathogenesis of CRC.
ABSTRACT
Objective To investigate the effect of sphingosine kinase 1 (SphK1) gene silence on the sensitivity to cisplatin (DDP) in RKO colon cancer cell line and the potential mechanism. Methods Targeted SphK1 gene lentivirus virus was constructed to infect RKO cells. The relative mRNA expression of SphK1 was detected by quantitative real-time PCR (qRT-PCR) and the protein level of SphK1 was determined by Western blotting. Then RKO cells were divided into three groups: down-regulated SphK1 group (shSphK1 group), negative control group (shControl group) and blank control group (control group). Cells of these groups were incubated for 0, 24, 48 and 72 hours with 0, 2, 4, 8, 16, 32 µg/mL DDP. After treatment, cell viability was evaluated by MTT assay. Cell apoptosis index was determined by TUNEL. The expressions of ki67, Bcl-2, caspase-9 and caspase-3 were tested by Western blotting. Results Down-regulation of SphK1 inhibited cell proliferation and enhanced apoptosis of RKO cells, expecially after exposed to DDP. Silence of SphK1 sensitized RKO cells to DDP in a concentration- and time-dependent manner. Cell proliferation of shSphK1 group was obviously reduced compared with control group or shControl group, and cell apoptosis rate of shSphK1 group significantly increased compared with control group or shControl group. Moreover, with the down-regulation of SphK1, the expressions of ki67 and Bcl-2 were depressed; the expressions of caspase-9 and caspase-3 were raised, especially after treated with DDP. Conclusion Down-regulation of SphK1 may decrease the expression of Bcl-2, increase the expressions of caspase-9 and caspase-3, inhibit cell proliferation, and promote cell apoptosis, thus improving chemosensitivity of colon cancer RKO cells to DDP.
Subject(s)
Cisplatin/pharmacology , Colonic Neoplasms/drug therapy , Colonic Neoplasms/genetics , Down-Regulation/genetics , Phosphotransferases (Alcohol Group Acceptor)/genetics , Cell Line, Tumor , HumansABSTRACT
Sphingosine kinase 1 (SphK1) plays an important role in colorectal carcinoma metastasis. However, whether SphK1 modulates epithelial-mesenchymal transition (EMT)-related marker expression and the underlying mechanisms remain unclear. In this study, in order to clarify this issue, we used various colorectal cancer (CRC) cell lines, Caco2, HT29, RKO and HCT116. Each of the cell lines was divided into 3 groups as follows: the control group, SKI-â ¡ (SphK1 inhibitor) group and PF-562271 [focal adhesion kinase (FAK) inhibitor] group. The migratory ability of the cells was examined by Transwell chamber assay. The mRNA and protein expression levels of SphK1, FAK (p-FAK), Slug, vimentin, N-cadherin and E-cadherin were detected by PCR and western blot analysis, respectively. The results revealed that the suppression of SphK1 reduced the cell migratory ability, and decreased the expression of Slug, vimentin and N-cadherin; however, the expression of E-cadherin was increased. Moreover, the inhibition of SphK1 reduced the expression of p-FAK. The inhibition of FAK (p-FAK) also decreased the cell migratory ability, and decreased the expression of Slug, vimentin and N-cadherin, whereas the expression of E-cadherin was increased. Thus, our data suggest that SphK1 modulates the expression of EMT-related markers and cell migration by regulating the expression of p-FAK in CRC cells. Thus, SphK1 may play a functional role in mediating the EMT process in CRC.
Subject(s)
Cell Movement/genetics , Colorectal Neoplasms/genetics , Epithelial-Mesenchymal Transition/genetics , Focal Adhesion Protein-Tyrosine Kinases/genetics , Gene Expression Regulation, Neoplastic , Phosphotransferases (Alcohol Group Acceptor)/genetics , Biomarkers , Cadherins/genetics , Cadherins/metabolism , Cell Line, Tumor , Colorectal Neoplasms/metabolism , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Humans , Phosphotransferases (Alcohol Group Acceptor)/metabolism , RNA, Messenger/genetics , Vimentin/genetics , Vimentin/metabolismABSTRACT
Objective To explore the clinical value of double-balloon enterocopy (DBE) in diagnosis of small intestinal diseases. Methods The clinical and endoscope image data of 231 patients with suspected small bowel disease who underwent DBE from January 2008 to May 2016 were analyzed. Result 231 patients received 257 times of DBE examination, 112 of them were performed by oral and 93 by anal route, 26 patients were underwent by both approaches. The detection rate of intestine diseases was 64.9% (150/231), include 33 cases (14.3%) of nonspecific enteritis, 27 cases (11.7%) of crohn's disease, 19 cases (8.2%) of ulcer, 13 cases (5.6%) of intestinal vascular malformation, 12 cases (5.2%) of small intestinal stromal tumor. The lesion detection rate in obscure abdominal pain and obscure gastrointestinal bleeding were 59.6% (62/104) and 67.0% (63/94). In all patients, there were 1 case of small bowel perforation, the remaining patients had no serious complications such as bleeding and perforation. Conclusion The positive detection rate of double-balloon enteroscopy examination is high, and the double-balloon enteroscopy examination is relatively safe. So, double-balloon enterscopy examination has high diagnostic value for detecting small intestine diseases.
ABSTRACT
Objective To explore the clinical value of double-balloon enterocopy (DBE) in diagnosis of small intestinal diseases. Methods The clinical and endoscope image data of 231 patients with suspected small bowel disease who underwent DBE from January 2008 to May 2016 were analyzed. Result 231 patients received 257 times of DBE examination, 112 of them were performed by oral and 93 by anal route, 26 patients were underwent by both approaches. The detection rate of intestine diseases was 64.9% (150/231), include 33 cases (14.3%) of nonspecific enteritis, 27 cases (11.7%) of crohn's disease, 19 cases (8.2%) of ulcer, 13 cases (5.6%) of intestinal vascular malformation, 12 cases (5.2%) of small intestinal stromal tumor. The lesion detection rate in obscure abdominal pain and obscure gastrointestinal bleeding were 59.6% (62/104) and 67.0% (63/94). In all patients, there were 1 case of small bowel perforation, the remaining patients had no serious complications such as bleeding and perforation. Conclusion The positive detection rate of double-balloon enteroscopy examination is high, and the double-balloon enteroscopy examination is relatively safe. So, double-balloon enterscopy examination has high diagnostic value for detecting small intestine diseases.
ABSTRACT
NIBP, a novel nuclear factor-κB (NF-κB)-inducing kinase (NIK) and IκB kinase ß (IKKß) binding protein, directly interacts with NIK and IKKß, and acts as the 'bridge' of the NFκB classical and alternative signaling pathways. However, its influence on epithelialmesenchymal transition markers in colon cancer remains to be fully elucidated. The aim of the present study was to investigate the roles of NIBP impacting on the expression of Ecadherin, CD44 and vimentin. In the present study, the associations between NIBP and Ecadherin, CD44 and vimentin in clinical samples were analyzed by making pairwise comparisons between normal colon tissue, nonmetastatic colon cancer tissue and metastatic colon cancer tissue. In in vitro experiments, after changing the expression of NIBP in cells, the protein expression levels of CD44, vimentin, Ecadherin were analyzed by western blot analysis. The results revealed that the protein expression levels of NIBP, CD44 and vimentin were markedly increased, and Ecadherin was markedly decreased, in metastatic colon cancer tissue compared with normal colon tissue and nonmetastatic colon cancer tissue. Upregulation of NIBP expression decreased the levels of Ecadherin, whereas the downregulation of NIBP increased Ecadherin levels, while no significant differences were observed in the levels of CD44 and vimentin. In addition, cells that were treated with the NFκB inhibitor, pyrrolidine dithiocarbamate (PDTC), also tended to exhibit increased levels of CD44 and vimentin expression in the NIBP upregulated expression group (29NIBP group) compared with the mock group, whereas the expression levels of Ecadherin, CD44 and vimentin were similar in the NIBP downregulated expression group (116NIBPmir group) and the HCT116 blank control group (116mock group) on treatment of the cells with tumor necrosis factorα. These findings indicated that NIBP, Ecadherin, CD44 and vimentin are possibly associated with metastasis in colon cancer. When the NFκB pathway is not subjected to any interventions, NIBP may predominantly regulate the NFκB classical pathway, rather than the alternative pathway. When the classical pathway was completely inhibited, NIBP was able to activate the NFκB alternative pathway. NIBP is therefore necessary for the interaction between the NFκB classical and alternative pathways. In conclusion, NIBP impacts on the expression levels of Ecadherin, CD44 and vimentin via the NFκB classical and alternative pathways. Therapeutic regimens for patients with colorectal cancer may comprise NIBP inhibitors in the future.
Subject(s)
Cadherins/biosynthesis , Carrier Proteins/biosynthesis , Colonic Neoplasms/metabolism , Gene Expression Regulation, Neoplastic , Hyaluronan Receptors/biosynthesis , NF-kappa B/metabolism , Neoplasm Proteins/metabolism , Vimentin/biosynthesis , Antigens, CD , Cadherins/genetics , Carrier Proteins/genetics , Cell Line, Tumor , Colonic Neoplasms/pathology , Female , Humans , Hyaluronan Receptors/genetics , Intercellular Signaling Peptides and Proteins , Male , NF-kappa B/genetics , Neoplasm Proteins/genetics , Vimentin/geneticsABSTRACT
Kinase suppressor of Ras 1 (KSR1) is a scaffold protein that modulates the activation of the oncogenic mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling pathway. Ginkgo biloba extract (EGb) 761 has been demonstrated to possess antitumor activity that may be related to the KSR1-mediated ERK signaling pathway. However, the roles and its underlying mechanism in gastric cancer are unclear. In the present study, 62 gastric cancer and matched normal tissues were exploited for immunohistochemistry and real-time fluorescent quantitative PCR detection. Results of the immunohistochemistry showed that the expression of ERK1/2 and p-ERK1/2 was correlated to the expression of KSR1 and p-KSR1 in the gastric cancer tissues, and the overexpression of KSR1, p-KSR1, ERK1/2 and p-ERK1/2 was significantly associated with histological grade, TNM stage, lymph node and distant metastasis. Compared with the normal tissues, the relative mRNA copy values of KSR1, ERK1 and ERK2 in the cancer tissues were 2.43 ± 0.49, 2.10 ± 0.44 and 3.65 ± 0.94. In addition, the expression of KSR1, p-KSR1, ERK1/2 and p-ERK1/2 in human gastric cancer multidrug resistant SGC-7901/CDDP cells was higher than that in the SGC-7901 cells as detected by the methods of immunocytochemistry and western blot analysis. EGb 761 not only suppressed expression of these proteins induced by cisplatin (CDDP) and etoposide in SGC-7901 cells, but also inhibited expression of these proteins in the SGC-7901/CDDP cells. Meanwhile, the proliferation-suppressing and apoptosis-inducing capacities of CDDP and etoposide were enhanced following combined treatment with EGb 761. Moreover, EGb 761 reduced the malondialdehyde (MDA) content and elevated the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the tumor cells. These results confirmed that activation of the KSR1-mediated ERK1/2 signaling pathway may contribute to tumorigenesis, metastasis and chemoresistance of human gastric cancer. EGb 761 enhanced the chemotherapy sensitivity and reversed the chemoresistance through suppression of the KSR1-mediated ERK1/2 pathway in gastric cancer cells, and the underlying mechanism may be related to its antioxidative activity.