MicroRNAs (miRs) serve an important role in cell differentiation, proliferation and apoptosis by negatively regulating gene expression at the transcriptional or post‑transcriptional level. EI24 autophagy associated transmembrane protein (EI24) is a tumor suppressor gene that serves an important role in the occurrence and development of digestive system tumors. However, little is known regarding the relationship between EI24 and the prognosis of patients with colorectal cancer (CRC). Our previous study confirmed EI24 as the target molecule of miR‑483, using reporter gene detection. Thus, the aim of the present study was to elucidate the effect of the abnormal expression of miR‑483 on the malignant phenotype of CRC through a series of cell function experiments and nude mice tumorigenicity experiments, and to determine the expression level of EI24, a downstream target gene of miR‑483, in CRC and its relationship with patient prognosis. In CRC tissues and cells, the expression level of miR‑483 was upregulated, while the expression level of EI24 was downregulated. Cell function tests such as MTT assay, cell cycle assay, colony formation assay, Migration and invasion assays and nude mice tumorigenicity experiments demonstrated that the overexpression of miR‑483 promoted the proliferation, invasion and metastasis of CRC. Moreover, the reverse transcription‑quantitative PCR results indicated that overexpression of miR‑483 inhibited the expression level of EI24. The relationship between the clinical data and immunohistochemical results from 183 patients with CRC and survival was examined. It was found that the expression level of EI24 was positively associated with the prognosis of patients. As a cancer‑promoting factor, miR‑483 enhances the proliferation, migration and invasion of CRC cells by reducing the expression level of EI24.

ObjectiveEarly screening of non‐alcoholic fatty liver disease (NAFLD) is of great significance for the early detection and intervention in NAFLD. MicroRNAs (miRNAs) are important regulators of metabolic diseases including NAFLD. The aim of this study was to investigate the association of serum miR‐29a‐c with NAFLD in a Chinese population.MethodsParticipants were divided into four groups based on the presence or absence of NAFLD and/or type 2 diabetes mellitus (T2DM). Quantitative polymerase chain reaction analysis was performed to quantify serum level of miR‐29a‐c. The association of miR‐29a‐c with NAFLD was evaluated.ResultsSerum miR‐29b, but not miR‐29a or miR‐29c, was positively associated with NAFLD (odds ratio [OR] 2.04 [1.16‐ 3.58], P = 0.013). Additionally, age, serum triglyceride and fasting plasma glucose (FPG) levels were independently associated with miR‐29b (β ± standard error [SE] = 0.004 ± 0.002, P = 0.019 for age; β ± SE = 0.110 ± 0.054, P = 0.042 for triglyceride; and β ± SE = 0.389 ± 0.161, P = 0.016 for FPG). MiR‐29b level was positively correlated with intrahepatic lipid content (β ± SE = 6.055 ± 2.630, P = 0.024) after adjusted for age, sex, and body mass index.ConclusionsSerum miR‐29b was associated with intrahepatic lipid content and NAFLD in a Chinese population‐based study.

LAMP2A is the key protein of chaperone-mediated autophagy (CMA), downregulation of LAMP2A leads to CMA blockade. CMA activation has been implicated in cancer growth, but the exact mechanisms are unclear. Elevated expression of LAMP2A was found in 8 kinds of tumors (n=747), suggesting that LAMP2A may have an important role in cancer progression. Unsurprisingly, LAMP2A knockdown in gastric cancer (GC) cells hindered proliferation, accompanied with altered expression of cell cycle-related proteins and accumulation of RND3/RhoE. Interactomic and KEGG analysis revealed that RND3 was a putative CMA substrate. Further study demonstrated that RND3 silencing could partly rescue the proliferation arrest induced by LAMP2A knockdown; RND3 was increased upon lysosome inhibition via both chemicals and LAMP2A-shRNA; Furthermore, RND3 could interact with CMA components HSPA8 and LAMP2A, and be engulfed by isolated lysosomes. Thus, constant degradation of RND3 by CMA is required to sustain rapid proliferation of GC cells. At last, the clinical significance of LAMP2A was explored in 593 gastric noncancerous lesions and 173 GC tissues, the results revealed that LAMP2A is a promising biomarker for GC early warning and prognosis of female GC patients.

Mesenchymal stem cells (MSCs) may influence the growth and metastasis of various human malignancies, including hepatocellular carcinoma (HCC). Therefore, the underlying mechanisms via which MSCs are able to affect malignancies require investigation. In the present study, the potential role of MSC in the angiogenesis of HCC was investigated. A total of 17 nude mouse models exhibiting human HCC were used to evaluate the effects of MSC on angiogenesis. A total of 8 mice were injected with human MSCs via the tail vein, and the remaining 9 mice were injected with phosphate-buffered saline as a control. A total of 35 days subsequent to the injection of MSCs, the microvessel density (MVD) of tumors was evaluated by immunostaining, using cluster of differentiation 31 antibody. The mRNA levels of transforming growth factor (TGF)β1, Smad2 and Smad7 were detected using reverse transcription-quantitative polymerase chain reaction. Protein expression levels of TGFβ1 and vascular endothelial growth factor (VEGF) in tumor tissues were analyzed using ELISA. Compared with controls, MVD in MSC-treated mice was significantly increased (28.00±9.19 vs. 18.11±3.30; P=0.006). The levels of TGFβ1 mRNA in the MSC-treated group were 2.15-fold higher compared with the control group (1.27±0.61 vs. 0.59±0.39; P=0.033), and MVD was higher in the group exhibiting increased TGFβ1 mRNA levels compared with the control group (26.50±9.11 vs. 19.44±6.14; P=0.038). In addition, a close correlation between the expression levels of TGFβ1 and VEGF was identified. The results of the present study suggested that MSCs may be capable of enhancing the angiogenesis of HCC, which may be partly due to the involvement of TGFβ1.

BACKGROUND:Esophageal squamous cell carcinoma (ESCC) remains one of the most lethal malignant tumors, and currently there is no effective ways to manage the late stage disease. Therefore clarifying the mechanisms underlying the development of ESCC is of great importance to develop novel therapeutic agents. The present study focuses on the interaction between neurotransmitter substance P (SP) together with its receptor NK-1R and ESCC progression. METHODS:The distribution of SP positive nerve fibers and expression of NK-1R were detected in ESCC tissue using immunohistochemistry. The effects of SP stimulation and blocking on the growth and migration of ESCC cells were measured by in vitro and in vivo assay. RESULTS:A higher density of SP positive nerve fibers and elevated NK-1R expression on ESCC cells were observed. More importantly, the SP positive fiber density was correlated with tumor size and lymph node metastasis. SP promoted ESCC cell proliferation and migration by modulation of intracellular calcium levels. CONCLUSION:NK-1R activation by SP stimulation promotes growth and migration of ESCC cells.

Adhesion of cancer cells to the extracellular matrix (ECM) causes a novel acquired chemotherapeutic drug‑resistant phenotype, referred to as cell adhesion-mediated drug resistance (CAM-DR). Our previous studies suggested that the adhesion molecule MGr1-Ag/37LRP may promote multidrug resistance in gastric cancer cells. Therefore, we investigated MGr1-Ag/37LRP binding-induced adhesion, and its role in CAM-DR. Initial studies revealed that, after adhesion to the ECM, the multidrug-resistant gastric cancer cell lines SGC7901/VCR and SGC7901/ADR showed significantly higher mean adhesive cell numbers than non‑resistant SGC7901 cells. We then investigated expression of MGr1-Ag/37LRP in gastric cancer cells adhering to laminin. Western blotting, RT-PCR and dual-luciferase reporter assays showed that laminin induced MGr1-Ag/37LRP expression and activity. In vitro and in vivo assays revealed that small interfering RNA against MGr1-Ag/37LRP significantly reduced CAM-DR in SGC7901/VCR cells. In vivo and in vitro analyses revealed that binding of MGr1-Ag/37LRP decreased intracellular drug accumulation by increasing P-glycoprotein and multidrug-associated protein expression, and inhibited drug-induced apoptosis by regulating Bcl-2 and Bax expression. These results indicate that MGr1-Ag/37LRP contributes to laminin-mediated CAM-DR in gastric cancer cells, and is a potentially effective target for reversing this phenomenon in gastric cancer.

It is well known that tumor microenvironment plays a vital role in drug resistance and cell adhesion‐mediated drug resistance (CAM‐DR), a form of de novo drug resistance. In our previous study, we reported that MGr1‐Ag/37LRP ligation‐induced adhesion participated in protecting gastric cancer cells from a number of apoptotic stimuli caused by chemotherapeutic drugs. Further study suggested that MGr1‐Ag could prompt CAM‐DR through interaction with laminin. However, the MGr1‐Ag‐initiated intracellular signal transduction pathway is still unknown. In this study, our experimental results showed that gastric cancer MDR cell lines mediated CAM‐DR through upregulation of Bcl‐2 by MGr1‐Ag interaction with laminin. Further study found that, as a receptor of ECM components, MGr1‐Ag/37LRP may activate the downstream signal pathway PI3K/AKT and MAPK/ERK through interaction with phosphorylated FAK. Moreover, the sensitivity to chemotherapeutic drugs could be significantly enhanced by inhibiting MGr1‐Ag/37LRP expression through mAbs, siRNA, and antisense oligonucleotide. According to these results, we concluded that the FAK/PI3K and MAPK signal pathway plays an important role in MGr1‐Ag‐mediated CAM‐DR in gastric cancer. MGr1‐Ag/37LRP might be a potential effective reversal target to MDR in gastric cancer.

Although epithelial barrier dysfunction in the gut has been extensively reported in ulcerative colitis (UC), the pathogenesis of this disease is not completely understood. In the present study, we investigated the role of miR-21 in regulating intestinal epithelial barrier function in UC. Colonic biopsies were obtained from 30 chronic UC patients and 30 healthy controls. Using real-time quantitative polymerase chain reaction (qRT-PCR), we found that both the mucosal and serum levels of miR-21 were upregulated in UC. In situ hybridization (ISH) analysis confirmed the accumulation of miR-21 in UC epithelia cells in vivo. Immunohistochemistry, Western Blot, qRT-PCR, and ultrastructural analyses further demonstrated that the overexpression of miR-21 in UC mucosa and Caco-2 cells impaired the integrity of the tight junctions, resulted in a decrease of the transepithelial electrical resistance (TER) and an increase of the inulin permeability. Furthermore, miR-21 induced the degradation of RhoB mRNA, which led to the depletion of RhoB and the impairment of tight junctions in intestinal epithelial cells.

MicroRNAs (miRNAs) play an essential role in colorectal cancer (CRC) development and progression. Aberrant miR‐625 expression has been reported in several cancers. However, the clinical significance of miR‐625 in human CRC has not been addressed.

BACKGROUND:Our previous study showed that BMP7 revealed significantly higher levels in esophageal squamous cell carcinoma (ESCC) tissues with lymph node metastasis compared with non-lymph node metastasis, using gene expression profiling assays. The roles of BMP7 in ESCC is not fully understood.AIM:The aim of this study was to investigate the effect of BMP7 on lymph node metastasis of ESCC and to explore its potential mechanism.METHODS:Expression of BMP7 in ESCC tissues was evaluated by immunohistochemistry. BMP7 were down-regulated by RNA interference. The protein and mRNA levels of BMP7 were detected by western blot and RT-PCR, respectively. High content screening and transwell assay were used to identify the metastatic ability of tumor cells.RESULTS:Positivity of BMP7 staining was 57.5 % in the tissues of primary carcinoma with lymph node metastasis compared to tissues without lymph node metastasis, and expression of BMP7 was significantly higher in the cell lines with highly metastatic capacity than that in the cell lines without metastatic ability. Suppression of endogenous BMP7 expression by siRNA in the highly metastatic cell lines resulted in significant reduction in ability of cell migration and invasion in both in vitro and in vivo studies. In addition, inhibition of BMP7 by siRNA also leads to up-regulation of E-cadherin and down-regulation of MMP-9 in the highly metastatic cell lines.CONCLUSIONS:These findings indicate that BMP7 modulates the expression of E-cadherin and MMP-9, and by which mechanism it may regulate cell migration and metastasis of ESCCs.

Background RhoE, an atypical Rho protein, is differently deregulated in some solid tumors, and there are conflicting data describing the role of RhoE in tumor cell migration and invasion. This study aimed to investigate RhoE expression in human colorectal cancer and its relationship with clinicopathological features and prognosis. Methods Colorectal cancer and adjacent normal tissues from 202 patients were examined by immunohistochemistry. Staining evaluation results were analyzed statistically in relation to various clinicopathological parameters, disease-free survival, and overall survival. RhoE expression was also investigated by immunohistochemistry in 80 node metastases and the corresponding primary lesions, and by Western blot test in six cancer and adjacent normal tissues. The relationship between RhoE and invasion was examined by transwell assay and Western blot test. Results The positive rate for RhoE in colorectal cancer was significantly higher than that of normal colorectal tissues. In colorectal cancer, RhoE expression was significantly correlated with depth of invasion, lymph node metastasis, and distant metastasis. Consistently, overexpression of RhoE in SW620 cells up-regulated vimentin, down-regulated E-cadherin, increased the expression of matrix metalloproteinase (MMP) 9, and enhanced cell invasion in vitro; in contrast, silencing of RhoE by a specific siRNA caused opposite effects. Most importantly, disease-free and overall survivals were significantly poorer for patients with RhoE-positive tumors than for those with RhoE-negative tumors. Conclusions These findings emphasize the positive role of RhoE in invasion and metastasis in human colorectal cancer. It could also serve as an independent prognostic marker in addition to the tumor, node, metastasis staging system.

Background: The epidermal growth factor receptor (EGFR) plays important roles in the development of gastric cancer. This study aims to analyze the prognostic value of EGFR in patients with gastric cancer.Methods: A meta-analysis is performed by searching Cochrane Library, PubMed, EMBASE and Science Direct databases from Jan 1970 to May 2013. Data are extracted from studies evaluating the survival of gastric cancer patients with either positive or negative EGFR expression. Pooled hazard ratios (HRs) and 95% confidence intervals (Cls) are calculated.Results: Totally 1600 cases of gastric cancer patients from five studies are subjected to final analysis. The HR of post-operational survival of patients with positive EGFR expression is 1.16 (95% CI: 0.94-1.43) as compared with those with negative expression, indicating that positive EGFR expression does not significantly predict the poor survival of gastric cancer.Conclusions: EGFR expression is not an independent predictor for the survival of gastric cancer patients. (C) 2013 Elsevier B.V. All rights reserved.

AIM:To investigate the role of interleukin (IL)-17 in small bowel allograft rejection.METHODS:We detected the expression of helper T cell 17 (Th17) cells in biopsy specimens from 3 cases of living small bowel transplantation in our department through immunofluorescence stain. We then established a rat heterotopic small bowel transplantation model. The rats were sacrificed on the 1(st), 2(nd), 3(rd), 5(th), and 7(th) d after small bowel transplantation. The degrees of transplantation rejection in rat intestine graft were examined through hematoxylin eosin (HE) stain, and the expression of Th17 cells in rat intestine graft were detected through immunofluorescence stain. In addition, the recipient rats undergoing intestinal transplantation were administrated with mouse-anti-rat IL-17 monoclonal antibody (mAb), and the survival of rats was analyzed. The recipient rats which received mouse-anti-rat IL-17 mAb treatment were sacrificed on the 1(st), 2(nd), 3(rd), 5(th), and 7(th) d after small bowel transplantation. The degrees of transplantation rejection and the expression of Th17 cells in rat intestine graft were detected through HE and immunofluorescence stain. The expression of IL-17, IL-1β, tumor necroses factor receptor-α (TNF-α), IL-6, and IL-8 in the intestine graft or serum were also detected.RESULTS:The expressions of Th17 cells ran parallel with the degree of acute rejection in human intestine grafts. The intestine graft rejection of rats was aggravated with prolonged duration after intestinal transplantation, and the expressions of Th17 cells were also correlated with the degree of acute rejection in rat intestine grafts. Administration of mouse-anti-rat IL-17 mAb prolonged the survival of rats after small bowel transplantation (P < 0.001). Furthermore, we found that the administration of mouse-anti-rat IL-17 mAb significantly decreased the intensity of CD4+IL-17+ Th17 cells in intestine grafts on the 2(nd), 3(rd), 5(th), and the 7(th) d (97.22 ± 4.05 vs 12.45 ± 2.02 on the 7(th) d, P < 0.0001), and suppressed the severity of acute rejection. The expression of IL-17 in the intestine graft declined after mouse-anti-rat IL-17 mAb administration on the 2(nd), 3(rd), 5(th), and the 7(th) d (0.88 ± 0.03 vs 0.35 ± 0.02 on the 7(th) d, P < 0.0001). We also detected the IL-17 serum level and found that the IL-17 level reduced from the 1(st) d to the 7(th) d (6.52 ± 0.18 ng/mL vs 2.04 ± 0.15 ng/mL on the 7(th) d, P < 0.0001). No significant difference in the level of IL-17 mRNA in the intestine graft was identified between the two groups. The levels of IL-1β, TNF-α, IL-6, and IL-8 mRNA in the intestine graft after the administration of mouse-anti-rat IL-17 mAb were also tested. We found that on the 3(rd), 5(th), and 7(th) d after intestinal transplantation, administration of mouse-anti-rat IL-17 mAb significantly inhibited the levels of IL-1β (12.11 ± 1.16 vs 1.27 ± 0.15 on the 7(th) d, P < 0.001), TNF-α (27.37 ± 2.60 vs 1.06 ± 0.26 on the 7(th) d, P < 0.001), IL-6 (21.43 ± 1.79 vs 1.90 ± 0.32 on the 7(th) d, P < 0.001), and IL-8 (20.44 ± 1.44 vs 1.34 ± 0.20 on the 7(th) d, P < 0.001) mRNA in the intestine graft.CONCLUSION:IL-17 may act as a promising and potent target for inhibiting acute rejection after small bowel transplantation.

BACKGROUND:The incidence of esophagogastric junctional adenocarcinoma is increasing, and the surgery is associated with high mortality and morbidity rates. This study aims to evaluate whether three-field minimally invasive surgery promotes outcome as compared with three-incision open surgery. METHODS:From January 1, 2009, to March 1, 2012, 114 consecutive patients with Siewert type I esophagogastric junctional adenocarcinoma were involved in this retrospective study. Patients were randomly assigned by a computer-generated randomization sequence to receive either three-incision open esophagectomy or minimally invasive esophagectomy. Details concerning patients and tumor characteristics, surgical procedures, and postoperative outcomes were collected and compared. RESULTS:Totally, 59 patients were involved in the open esophagectomy group and 55 in the minimally invasive esophagectomy group. The incidence of pulmonary morbidity (9.09% versus 28.81%) and vocal cord paralysis (0% versus 15.25%) in the minimally invasive group was significantly less than that in the open esophagectomy group. Furthermore, the postoperative life quality in the minimally invasive group was better than that in the open group. Survival at 2 years was 83.6% for the minimally invasive group (46 of 55 patients) and 79.7% for the open esophagectomy group (47 of 59 patients). CONCLUSIONS:Minimally invasive esophagectomy could lead to a significant improvement of the short-term benefits for patients with Siewert type I esophagogastric junctional adenocarcinoma.

MicroRNAs (miRNAs) are small non-coding RNA molecules that function as negative regulators of gene expression. Common genetic variants (single nucleotide polymorphisms, SNPs) in miRNA genes may alter their expression or maturation resulting in varied functional consequences. Until now, several studies had evaluated the association between the polymorphisms in the hsa-miR-196a2 rs11614913 and cancer risk in diverse populations and in multiple types of cancer, with contradictory outcomes. Therefore, here we performed a meta-analysis to address the association between this polymorphism and cancer risk. A total of nine studies involving 6,540 cases and 7,562 controls were retrieved based on PubMed. Our analysis demonstrated that hsa-miR-196a2 rs11614913 CC genotype significantly increased the cancer risk in homozygote comparison model compared to TT genotype (OR=1.18; 95% CI, 1.01-1.68). Moreover, significant association of this polymorphism with breast cancer was found based on homozygote comparison model (OR=1.30; 95% CI, 1.01-1.26) and dominant model (OR=1.11; 95% CI, 1.01-1.23). In addition, hsa-miR-196a2 rs11614913 CC genotype was significantly associated with cancer risk in Chinese and Indian (OR=1.21; 95% CI, 1.05-1.40), but not in Caucasians (OR=1.03; 95% CI, 0.89-1.19). Taken together, our results indicate that the polymorphism of hsa-miR-196a2 rs11614913 is associated with cancer susceptibility, especially with breast cancer and in Chinese and Indian populations.