Effects of miR-362 in Regulating the Proliferation, Invasion and Apoptosis of Gastric Cancer by Inhibiting the Expression of Tumor-Promoting Factor PRDM2.

Purpose - The high expression of positive regulatory domain zinc finger protein2 (PRDM2) is an important factor in inducing the formation and progression of gastric cancer. The current study was performed to explore the effect of micro-RNA-362 (miR-362) targeting PRDM2 on the proliferation and apoptosis of gastric cancer cells. Methods - The expression of miR-362 in gastric adenocarcinoma and normal gastric mucosa was detected by real-time fluorescence quantitative PCR (qPCR), and the expression of PRDM2 in gastric adenocarcinoma was detected by im-munohistochemical method. Gastric cancer cell line MGC-803 and human normal gastric mucosal epithelial cell line (GES-1) were selected for study. Blank control group, empty vector transfection group, miR-362 transfection group, and miR-362 and PRDM2 co-transfection group were established. CCK-8 assay was utilized to detect cell activity, flow cytometry was used to detect cell cycle and apoptosis, and invasion capability of cells was observed through transwell experiment. The expression of Cyclin D1 and Caspases-3 was detected by western blot method. Results - The expression of miR-362 in gastric adenocarcinoma was significantly lower than that in normal gastric mucosa. The expression of miR-362 in gastric adenocarcinoma was significantly different in the maximum diameter, depth of invasion, and different TNM stages. The expression of miR-362 was linked to prognosis. In addition, there was a negative correlation between miR-362 and PRDM2. The expression of miR-362 in gastric cancer cell line MGC-803 was significantly lower than that in GES-1. Compared with the blank control group and empty vector transfection group, the proliferation level of gastric cancer cells in miR-362 transfection group was remarkably decreased, the cells in S phase and G2/M phase were significantly decreased, the capability for invasion was evidently decreased, while the apoptosis was significantly increased. The expression of Caspases-3 increased significantly and the expression of Cyclin D1 decreased significantly. MiR-362 and PRDM2 co-transfection groups could reverse the abovementioned expression levels. Conclusion - MiR-362 can regulate the proliferation, invasion and apoptosis of gastric cancer by inhibiting the expression of tumor-promoting factor PRDM2. The expression of miR-362 in gastric adenocarcinoma is significantly decreased, which can regulate the formation and development of gastric adenocarcinoma by promoting the expression of PRDM2. Moreover, low expression of miR-362 in gastric adenocarcinoma is an important risk factor for tumor progression and poor prognosis.

Pharmacokinetics and Tissue Distribution of Anwuligan in Rats after Intravenous and Intragastric Administration by Liquid Chromatography-Mass Spectrometry.

Anwuligan, a natural 2,3-dibenzylbutane lignan from the nutmeg mace of Myristica fragans, has been proved to possess a broad range of pharmacological effects. A rapid, simple, and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been established and successfully applied to the study of pharmacokinetics and tissue distribution of anwuligan after intravenous or intragastric administration. Sample preparation was carried out through a liquid-liquid extraction method with ethyl acetate as the extraction reagent. Arctigenin was used as the internal standard (IS). A gradient program was employed with a mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile. The mass spectrometer was operated in a positive ionization mode with multiple reaction monitoring. The transitions for quantification were m/z 329.0→205.0 for anwuligan and m/z 373.0→137.0 for IS, respectively. Calibration curves were linear over the ranges of 0.5-2000 ng/mL for both plasma samples and tissue samples (r > 0.996). The absolute bioavailability is 16.2%, which represented the existing of the obvious first-pass effect. An enterohepatic circulation was found after the intragastric administration. Anwuligan could be distributed rapidly and widely in different tissues and maintained a high concentration in the liver. The developed and validated LC-MS/MS method and the pharmacokinetic study of anwuligan would provide reference for the future investigation of the preclinical safety of anwuligan as a candidate drug.

The Combination of Diffusion- and T2-Weighted Imaging in Predicting Deep Myometrial Invasion of Endometrial Cancer: A Systematic Review and Meta-Analysis.

OBJECTIVE: The aim of this study was to explore the diagnostic difference among the combination of diffusion-weighted imaging (DWI) and T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), and dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) in predicting deep myometrial invasion of endometrial cancer. METHODS: A structured search was conducted to identify published studies between January 2005 and April 2014, which assessed depth of myometrial invasion in endometrial cancer by using DCE-MRI or DWI or DWI-T2WI. RESULTS: A total of 15 studies were included. Significant difference was found between DWI-T2WI and DWI in pooled specificity, and also in comparison between DCE-MRI and DWI-T2WI (P < 0.05). In summary, receiver operating characteristic analysis, area under the curve for DWI-T2WI, DWI, and DCE-MRI were 0.94, 0.90, and 0.93, respectively. CONCLUSIONS: Diffusion-weighted imaging-T2WI can improve diagnostic performance in comparison with DWI alone. Meanwhile, DWI-T2WI performs better than DCE-MRI in predicting myometrial invasion of endometrial cancer. It may be an alternative for DCE-MRI in presurgical staging of endometrial cancer.

Fluorodeoxyglucose positron emission tomography and chemotherapy-related tumor marker expression in non-small cell lung cancer.

BACKGROUND: The chemotherapy resistance of non-small cell lung cancer (NSCLC) remains a clinic challenge and is closely associated with several biomarkers including epidermal growth factor receptor (EGFR) ( Drugs 72(Suppl 1):28-36, 012.), p53 ( Med Sci Monit 11(6):HY11-HY20, 2005.) and excision repair cross complementing gene 1 (ERCC1) ( J Thorac Oncol 8(5):582-586, 2013.). Fluorodeoxyglucose positron emission tomography (FDG-PET) is the best non-invasive surrogate for tumor biology with the maximal standardized uptake values (SUVmax) being the most important paradigm. However, there are limited data correlating FDG-PET with the chemotherapy resistant tumor markers. The purpose of this study was to determine the correlation of chemotherapy related tumor marker expression with FDG-PET SUVmax in NSCLC. METHODS: FDG-PET SUVmax was calculated in chemotherapy naïve patients with NSCLC (n=62) and immunohistochemical analysis was performed for EGFR, p53 or ERCC1 on the intraoperative NSCLC tissues. Each tumor marker was assessed independently by two pathologists using common grading criteria. The SUVmax difference based on the histologic characteristics, gender, differentiation, grading and age as well as correlation analysis among these parameters were performed. Multiple stepwise regression analysis was further performed to determine the primary predictor for SUVmax and the receiver operating characteristics (ROC) curve analysis was performed to detect the optimized sensitivity and specificity for SUVmax in suggesting chemotherapy resistant tumor markers. RESULTS: The significant tumor type (P=0.045), differentiation (P=0.021), p53 (P=0.000) or ERCC1 (P=0.033) positivity dependent differences of SUVmax values were observed. The tumor differentiation is significantly correlated with SUVmax (R=-0.327), tumor size (R=-0.286), grading (R=-0.499), gender (R=0.286) as well as the expression levels for p53 (R= -0.605) and ERCC1 (R=-0.644). The expression level of p53 is significantly correlated with SUVmax (R=0.508) and grading (R=0.321). Furthermore, multiple stepwise regression analysis revealed that p53 expression was the primary predictor for SUVmax. When the cut-off value of SUVmax was set at 5.15 in the ROC curve analysis, the sensitivity and specificity of SUVmax in suggesting p53 positive NSCLC were 79.5% and 47.8%, respectively. CONCLUSION: The current study suggests that SUVmax of primary tumor on FDG-PET might be a simple and good non-invasive method for predicting p53-related chemotherapy resistance in NSCLC when we set the cu-off value of SUVmax at 5.15.

Progress in Pretreatment and Analysis of Cephalosporins: An Update Since 2005.

Cephalosporins, one of the classic ß-lactam antibiotics, has been widely concerned by global population as the most commonly used broad-spectrum antibiotic. Serious health hazards are possible to be posed by quality control problems caused by its unstable structure, as well as food and environmental pollution introduced by improper use. Given the reasons above, the sensitive and valid methods for monitor and determination of cephalosporins in different matrices are urgently required. This review aims to provide a comprehensive overview on the current status of the pretreatment and analysis methods of cephalosporins in bulk drug, pharmaceutics, animal-derived foodstuffs including eggs, milk, meat, environment samples, and biological samples. Pretreatment methods including simple steps (protein precipitation, centrifugation, filtration), liquid-liquid extraction, solid phase extraction, QuECHERS, and detection methods covering LC, LC-MS/MS, voltammetric sensor, capillary electrophoresis spectroscopy, biological methods from January 2005 to October 2018 are updated, elaborated and compared here. Moreover, advanced materials and prospects for development are discussed. HighlightsDetermination of cephalosporins in different newly found matrix are represented.Comparisons between different mass analyzers and progress in HRMS methods are in detailed.Optimization of experimental conditions are discussed.Newly emerged eco-friendly methods are introduced.

A meta-analysis of the accuracy of prostate cancer studies which use magnetic resonance spectroscopy as a diagnostic tool.

OBJECTIVE: We aimed to do a meta-analysis of the existing literature to assess the accuracy of prostate cancer studies which use magnetic resonance spectroscopy (MRS) as a diagnostic tool. MATERIALS AND METHODS: Prospectively, independent, blind studies were selected from the Cochrane library, Pubmed, and other network databases. The criteria for inclusion and exclusion in this study referenced the criteria of diagnostic research published by the Cochrane center. The statistical analysis was adopted by using Meta-Test version 6.0. Using the homogeneity test, a statistical effect model was chosen to calculate different pooled weighted values of sensitivity, specificity, and the corresponding 95% confidence intervals (95% CI). The summary receiver operating characteristic (SROC) curves method was used to assess the results. RESULTS: We chose two cut-off values (0.75 and 0.86) as the diagnostic criteria for discriminating between benign and malignant. In the first diagnostic criterion, the pooled weighted sensitivity, specificity, and corresponding 95% CI (expressed as area under curve [AUC]) were 0.82 (0.73, 0.89), 0.68 (0.58, 0.76), and 83.4% (74.97, 91.83). In the second criterion, the pooled weighted sensitivity, specificity, and corresponding 95% CI were 0.64 (0.55, 0.72), 0.86 (0.79, 0.91) and 82.7% (68.73, 96.68). CONCLUSION: As a new method in the diagnostic of prostate cancer, MRS has a better applied value compared to other common modalities. Ultimately, large scale RCT (randomized controlled trial) randomized controlled trial studies are necessary to assess its clinical value.

Technetium-99m-hexakis-2-methoxyisobutylisonitrile scintigraphy and multidrug resistance-related protein expression in human primary lung cancer.

OBJECTIVE: The occurrence of multidrug resistance (MDR) is a major cause of resistance to chemotherapeutic agents in patients with lung cancer, in part owing to the overexpression of MDR-related proteins. Technetium-99m-hexakis-2-methoxyisobutylisonitrile ((99m)Tc-MIBI) has been shown to be a substrate for some MDR-related proteins. The aim of this study is to evaluate the role of (99m)Tc-MIBI scintigraphy for functional imaging of MDR-related protein phenotypes. METHODS: To determine the correlation between (99m)Tc-MIBI scintigraphy and the expression level of P-glycoprotein (Pgp), multidrug-resistance protein (MRP), and glutathione-S-transferase Pi (GSTpi), 26 patients (17 men and 9 women, median age 57.5 years) with primary lung cancer were investigated. Following intravenous administration of 925 MBq (99m)Tc-MIBI, single-photon emission computed tomography (SPECT) and computed tomography (CT) were performed at 15 min and 2 h. On the basis of the fused images, tumor to background (T/B) ratio of both early and delayed images, and washout rate (WR%) of (99m)Tc-MIBI were calculated. The immunohistochemical staining of Pgp, MRP, and GSTpi was performed, and the expression level was semiquantitated using a pathoimage analysis system. The imaging results were compared with the status of Pgp, MRP, and GSTpi expression. RESULTS: The WR% of (99m)Tc-MIBI showed a significant positive correlation with Pgp expression (r = 0.560, P = 0.003), as no correlation was observed between WR% and MRP or GSTpi (r = 0.354, P = 0.076; r = 0.324, P = 0.106). Neither early T/B nor delayed T/B correlated with the expression level of Pgp, MRP, and GSTpi. WR%, Pgp, and GSTpi expression showed significant differences between squamous cell carcinoma (group A) and adenocarcinoma (group B). There was no significant difference among Pgp, MRP, and GSTpi expression levels in any cases (P > 0.05). CONCLUSIONS: Our data confirmed that (99m)Tc-MIBI scintigraphy is useful for determining the MDR caused by Pgp in patients with primary lung cancer.

The utility of measuring the apparent diffusion coefficient for peritumoral zone in assessing infiltration depth of endometrial cancer.

BACKGROUND: The invasion depth of endometrial cancer is one of the most important prognosis factors. The aim of the current study was to investigate the diagnostic value of the apparent diffusion coefficient (ADC) of the peritumoral zone for assessing the infiltration depth of endometrial cancer. METHODS: An institutional review board approved this prospective study, and all study participants provided informed consent. A total of 58 patients (mean age 54 ± 8.3 years, range 34-69 years) with endometrial cancer were prospectively enrolled. Two radiologists assessed all preoperative magnetic resonance images with T1, T2, and diffusion-weighted imaging, and determined the location of the deepest invasion of the tumor. The peritumoral zone was defined as a 5-mm-thick zone surrounding and adjacent to the cancerous endometrium. The mean ADC (ADCm) values of the tumor and the peritumoral zone were measured. Sensitivity, specificity, positive and negative predictive values, and the area under the receiver operating characteristic curve (Az) were calculated for visual inspection, and an ADC cutoff value for the peri-endometrial zone was determined for predicting the myometrial invasion depth. RESULTS: The ADCm values of tumors and peritumoral zones were 0.83 × 10- 3 mm2/sec and 1.06 × 10- 3 mm2/sec, respectively. There was no significant difference between the ADCm values of the tumors in the superficial and deep myometrial invasion groups (P > 0.05). However, the ADCm value at the peritumoral zone in the deep myometrial invasion group (1.23 × 10- 3 mm2/sec) significantly differed from that in the superficial myometrial invasion group (0.99 × 10- 3 mm2/sec) (p = 0.005). In assessments of deep myometrial invasion, the sensitivity, specificity, negative predictive value, and positive predictive value were 0.58, 0.93, 0.84, and 0.77, respectively, for the ADCm cutoff value of the peritumoral zone, and 0.71, 0.80, 0.87, and 0.60. respectively, for visual inspection. The accuracy of myometrial invasion depth assessment using the ADCm cutoff value and visual inspection were 83 and 78%, respectively. The Az for both was 0.76. CONCLUSION: ADCm at the peritumoral zone can predict deep myometrial invasion of endometrial cancer. This value can therefore enhance confidence in preoperative endometrial cancer evaluation, and when tailoring surgical approaches.

Intercellular imaging by a polyarginine derived cell penetrating peptide labeled magnetic resonance contrast agent, diethylenetriamine pentaacetic acid gadolinium.

BACKGROUND: The cellular plasma membrane represents a natural barrier to many exogenous molecules including magnetic resonance (MR) contrast agent. Cell penetrating peptide (CPP) is used to internalize proteins, peptides, and radionuclide. This study was undertaken to assess the value of a new intracellular MR contrast medium, CPP labeled diethylenetriamine pentaacetic acid gadolinium (Gd-DTPA) in molecular imaging in vitro. METHODS: Fluorescein-5-isothiocyanate (FITC) and Gd-DTPA respectively labeled with CPP (FITC-CPP, Gd-DTPA-CPP) were synthesized by the solid-phase method. Human hepatic cancer cell line-HepG2 was respectively stained by FITC-CPP and FITC to observe the uptake and intracellular distribution. HepG2 was respectively incubated with 100 nmol/ml Gd-DTPA-CPP for 0, 10, 30, 60 minutes, and imaged by MR for studying the relationship between the incubation time and T(1)WI signal. The cytotoxicity to NIH3T3 fibroblasts cells was measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide reduction assay (MTT). RESULTS: The molecular weights of CPP labeled imaging agents, which were determined by MALDI mass spectrometry (FITC-CPP MW = 2163.34, Gd-DTPA-CPP MW = 2285.99), were similar to the calculated molecular weights. Confocal microscopy suggested HepG2 translocated FITC-CPP in cytoplasm and nucleus independent with the incubation temperature. MR images showed HepG2 uptaken Gd-DTPA-CPP had a higher T(1) weighted imaging (T(1)WI) signal, and that the T(1)WI signal intensity was increasing in a time-dependent manner (r = 0.972, P = 0.001), while the signal intensity between the cells incubated by Gd-DTPA for 60 minutes and the controlled cells was not significantly different (P = 0.225). By MTT, all concentrations from 50 nmol/ml to 200 nmol/ml had no significant (F = 0.006, P = 1.000) effect on cell viability of mouse NIH3T3 fibroblasts, compared with the control group. CONCLUSIONS: The newly constructed CPP based on polyarginine can translocate cells by carrying FITC and MR contrast agent Gd-DTPA, and the intracellular concentrations are readily detectable by MR imaging, suggesting a new way for MR molecular imaging.

[Characteristics and in vitro imaging study of matrix metalloproteinase-2 targeting activable cell-penetrating peptide].

OBJECTIVE: To study invasively imaging MMP2-positive tumor cell by paramagnetic Gadolinium or fluorescein carried by a activable cell penetrating peptides. METHODS: To label Fluorescein-5-isothiocyanate (FITC) and gadopentetate with the activable cell-penetrating peptides by a solid-phase synthesis method. Identification by TOF mass spectrum (TOF-MS). Isoelectric point of the activable cell penetrating peptides is determined by disc electrophoresis. T1 relacion of gadopentetate labeled with the activable cell-penetrating peptides (B) in water were determined on 400 MHZ NMR. Human lung cancer cell lines: A549 were respectively stained by FITC labeled with ACPPs (A) or FITC alone. MMP2 expression and activity were determined by zymography. T1WI signal of A549 incubated with 120 nmol/ml B or Diethylenetriaminepentaacetic Acid-Gadolinium (Gd-DTPA) for different times were obtained by 1.5T MRI. The location of B in A549 was detected by Transmission Electron Microscopy. Visualization analysis and half-quantitative analysis were used to determine the signal characteristics. The ANOVA analysis and the paired samples t test were performed by SPSS 13.0. RESULTS: MALDI TOF-MS molecular weigh of A and B respectively is 3789.74 and 3911.93. Isoelectric point is 11.005T1 Relacion of 0.5 mmol/L Gd-DTPA and B at 17 degrees C respectively is (0.052 +/- 0.01) sec and (0.050 +/- 0.001) sec. Fluorescein uptake assays showed that A translocated into A549 but would be inhibited by MMP2 antibody. Zymography showed both active-MMP2 (67000) and pro-MMP2 (72000) expressed byA549. MR imaging showed A549 incubating with B had a high T(1) signal, and the signal of A549 incubating with Gd-DTPA is similar with that of the control group. Furthermore, ANOVA suggested that the T(1) signal intensity of A549 incubating with B was effected by incubating time (F = 267.569, P < 0.001) and increasing in a time-dependent fashion at the observed time. There is no difference between the T(1) signal intensity of A549 incubating with Gd-DTPA and the control group (P > 0.05). TEM showed A in cytoplasm and nucleus. CONCLUSION: The study in vitro suggests that the MMP-2 activable cell-penetrating peptides bearing contrast media can detect the MMP2-positive tumor cell.

Combined subjective and quantitative analysis of magnetic resonance images could improve the diagnostic performance of deep myometrial invasion in endometrial cancer.

PURPOSE: To evaluate whether the combination of subjective magnetic resonance imaging (MRI) and quantitative analysis by using the exponential ADC (EADC) value of the peri-endometrial zone can improve the diagnostic performance of deep myometrial invasion in endometrial cancer patients. MATERIALS AND METHODS: We prospectively evaluated 111 patients with either cervical cancer (normal endometria group) or endometrial cancer (endometrial cancer group). Two radiologists assessed all preoperative MR images with T1, T2, and diffusion-weighted imaging. The EADC value of the peri-endometrial zone was measured. Sensitivity, specificity, positive and negative predictive values, and the area under the receiver operating characteristic curve (Az) were calculated for Subjective MRI, an EADC cutoff value of the peri-endometrial zone and the combination of the two methods in assessing the prediction of deep myometrial invasion. RESULTS: Specificity for EADC cutoff of the peri-endometrial zone was higher (0.93) than for Subjective MRI (0.80), as were the positive predictive values (EADC, 0.79; visual, 0.60). Sensitivity for the combined test was higher (0.88) than for Subjective MRI (0.71) and the EADC cutoff value (0.65), as were the negative predictive values (the combined test, 0.94; vs. EADC, 0.79; vs. Subjective MRI, 0.60). There were no differences in Az between the three methods (P>0.05), but the combined test had the highest Az. CONCLUSIONS: Combined with conventional Subjective MRI, calculating EADC value of the peri-endometrial zone could improve the accuracy of preoperative assessment of deep myometrial invasion in endometrial cancer patients, and maybe helpful in tailoring a surgical approach for intervention.

SUVmax of 18F-FDG PET/CT correlates to expression of major chemotherapy-related tumor markers and serum tumor markers in gastric adenocarcinoma patients.

The expression of P53 was previously found by us significantly correlated with maximal standardized uptake value (SUVmax) in non-small cell lung cancer (NSCLC) patients. Hence, the aim of this study was to clarify the relationship between SUVmax and the status of the chemotherapy-related tumor marker expression or serum tumor markers in gastric adenocarcinoma patients. Sixty-four gastric adenocarcinoma patients who underwent 18F-FDG PET/CT prior to treatment were enrolled in this study. Immunohistochemistry was performed to detect changes of Her-2, P53 and Survivin in lesions, and electrochemiluminescence (ECL) method was used to quantify expression of serum CA72-4, CA19-9 and CEA of these patients. Then, the relationships between these parameters above were assessed by Spearman correlation analysis. Also, receiver-operating characteristic (ROC) curve was performed to determine the best cut-off value of SUVmax for suggesting chemotherapy resistant tumor markers. Besides, we identified a linear correlation to estimate the equations between SUVmax and the serum tumor markers. Our results showed that higher SUVmax was detected in patients with positive expression of Her-2 and P53, compared with negative groups. The Spearman correlation analysis showed that SUVmax was associated with Her-2 or P53 with the moderate relevant Pearson correlation coefficient. ROC curve analysis showed that the sensitivity and specificity of SUVmax for suggesting Her-2 or P53-positive, when the cut-off value of SUVmax was set at 3.25 or 5.45, respectively. Moreover, the relationship between SUVmax and serum tumor markers were analyzed by linear correlation analysis, and serum CA72-4 and CA19-9 could be used as independent parameters to establish an equation for SUVmax by the linear regression models. These results suggested that SUVmax of 18F-FDG PET/CT could be used to predict and evaluate Her-2 or P53 related chemotherapy resistance of gastric adenocarcinoma patients. However, before PET/CT scanning, serum tumor markers could be used to calculate the SUVmax approximately.

Prognostic role of pretreatment neutrophil to lymphocyte ratio in breast cancer patients: A meta-analysis.

BACKGROUND: Inflammation and cancer are closely related to each other. As a parameter that can reflect inflammation and host immune reaction, elevated blood neutrophil to lymphocyte ratio (NLR) has been confirmed to be correlated with poor prognosis in a variety of cancers. However, this remains controversial in breast cancer. Thus, we performed this updated meta-analysis to further clarify whether high NLR could be a predictor of survival in breast cancer patients. METHODS: We searched on PubMed Database and Cochrane Library. Overall survival (OS), disease-free survival (DFS), and cancer-specific survival were used as outcome events, and hazard ratio (HR) was chosen as the parameter to evaluate the correlation. RESULT: Eighteen eligible studies were involved in this meta-analysis. The synthesized analysis demonstrated that elevated NLR was associated with poor DFS [HR = 1.72, 95% confidence interval (95% CI) = 1.30-2.27], OS (HR = 1.87, 95% CI = 1.41-2.48), and cancer-specific survival (HR = 2.09, 95% CI = 1.04-4.21). The correlation was stronger in triple-negative breast cancer (TNBC) (OS: HR = 2.58, 95% CI = 1.63-4.06; DFS: HR = 3.51, 95% CI = 1.97-6.24). CONCLUSION: Higher NLR was correlated to poor prognosis of breast cancer patients. As a clinical parameter that we can easily obtain, NLR might be a potential predictor in patients' survival to assist with physicians' treatment decisions.

High tumor-associated macrophages infiltration is associated with poor prognosis and may contribute to the phenomenon of epithelial-mesenchymal transition in gastric cancer.

BACKGROUND: Recent studies show that epithelial-mesenchymal transition (EMT) and tumor-associated macrophages (TAMs) contribute to the progression and poor prognosis of carcinoma through multiple mechanisms. Both inflammation and changing of epithelium have a close relationship with tumorigenesis of gastric cancer. However, the relevance between EMT and TAMs is still unclear in gastric cancer and needs more scientific research. This study is designed to explore the relationship between EMT and TAMs in gastric cancer. MATERIALS AND METHODS: Immunohistochemistry was used to detect the expression of EMT-related proteins and TAM markers in cancer tissues and normal gastric tissues. RESULTS: High levels of EMT and TAMs infiltration are related to aggressive features and independent prognostic factors in gastric cancer, respectively. In addition, expression of the two indicators is associated with expression of transforming growth factor-ß1 (TGF-ß1). Infiltration of TAMs is also associated with EMT-related marker in gastric cancer. CONCLUSION: Our results suggest that high levels of EMT and TAMs infiltration are related to aggressive features and independent prognostic factors in gastric cancer, respectively. A correlation was found between EMT- and TAM-related indicators, which may be associated with TGF-ß signaling pathway. The level of TAMs infiltration plays an important role in gastric cancer, the markers of which can be used as prognostic indicators.

Expression of RAP1B is associated with poor prognosis and promotes an aggressive phenotype in gastric cancer.

Metastasis is the major cause of death among gastric cancer (GC) patients, and altered expression of Ras-related protein RAP1B is associated with cancer development. The present study assessed RAP1B expression ex vivo and the effect of hypoxia­induced RAP1B overexpression on the promotion of the metastatic potential of GC cells in vitro. Immunohistochemistry was used to detect the expression of RAP1B and hypoxia­inducible factor-1α (HIF-1α) in 178 GC tissue specimens. GC cell lines were used to assess the effects of hypoxia and RAP1B knockdown with RAP1B small interfering RNA (siRNA). Tumor cell viability was detected by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, invasion capacity was evaluated by a Transwell assay, and gene expression was determined by qRT-PCR and western blotting. The data showed that expression levels of RAP1B and HIF-1α proteins were high in the GC tissue specimens, and RAP1B expression was significantly associated with tumor-node-metastasis (TNM) stage and tumor size, while HIF-1α expression was significantly associated with TNM stage, Borrmann type and tumor differentiation. Moreover, RAP1B expression was associated with HIF-1α expression (r=0.547, P<0.001). The expression of RAP1B and HIF-1α proteins was associated with a shorter overall survival of patients according to the univariate analysis (log-rank test, P<0.01), and RAP1B expression and TNM stage were independent prognostic predictors in patients using a multivariate analysis (P<0.001). In vitro, hypoxia induced the invasion of GC cells and the expression of RAP1B and HIF-1α (P<0.05); whereas knockdown of RAP1B expression using siRNA inhibited the tumor cell invasion capacity even under hypoxic culture conditions (P<0.05). In conclusion, the protein expression of RAP1B and HIF-1α contributed to GC malignant behavior and poor prognosis. Future studies will evaluate whether targeting RAP1B expression can be used as a novel strategy to control GC or as a biomarker for prognosis prediction.

FT-IR spectroscopic analysis of normal and cancerous tissues of esophagus.

AIM: To investigate the special Fourier transform infrared spectroscopy (FT-IR) spectra in normal and cancerous tissues of esophagus. METHODS: Twenty-seven pairs of normal and cancerous tissues of esophagus were studied by using FT-IR and the special spectra characteristics were analyzed in different tissues. RESULTS: Different spectra were found in normal and cancerous tissues. The peak at 1 550/cm was weak and wide in cancerous tissues but strong and high in normal tissues. The ratio of I 1 647/I 1 550 was 2.0 in normal tissues and 2.36 in cancerous tissues (P<0.05). The ratio of I 1 550/I 1 080 was 4.5 in normal tissues and 3.4 in cancerous tissues (P<0.01). The peak at 1453 /cm was higher than at 1 402/cm in normal tissue and lower than at 1 402/cm in cancerous tissues. CONCLUSION: The results indicate that FTIR may be used in clinical diagnosis.

[Effect of low dose propofol on biological behavior of esophageal squamous cell carcinoma cell line Eca109].

AIM: To detect the effect of low dose propofol on the proliferation, apoptosis, migration, and invasion of esophageal squamous cell carcinoma cell line Eca109. METHODS: The proliferation, apoptosis, migration, and invasion of esophageal squamous cell carcinoma cell line Eca109 were detected by MTT assay, flow cytometry, transwell assay respectively. The effect of low dose propofol on expression of heme oxygenase-1 (HO-1) was confirmed by Real-time quantitative PCR. RESULTS: Low dose propofol could inhibit the proliferation, migration, invasion and promate the apoptosis of esophageal squamous cell carcinoma cell line Eca109. And low dose propofol increased the expression of HO-1 mRNA in a dose-dependment manner. CONCLUSION: Low dose propofol affects the biological behavior of esophageal squamous cell carcinoma cell line Eca109, which has a relationship with increasing the expression of HO-1.

[Differential expression of hsa-miR-126 and hsa-miR-518b in esophageal squamous carcinoma].

OBJECTIVE: To identify the differentially expressed microRNAs (miRNAs) between esophageal squamous carcinoma (ESC) and adjacent non-tumorous tissue (NT). METHODS: The expression levels of the miRNAs were detected in 3 fresh ESC and NT samples by hybridization with miRNAs microarray chip. Real-time quantitative RT-PCR was performed to confirm the results of the microarray analysis. The expressions of hsa-miR-126 and hsa-miR-518b in ESC were validated by real-time quantitative RT-PCR in another independent 15 matched samples. RESULTS: A total of 11 miRNAs exhibited differential expressions in ESC samples as compared to their expressions in the NT samples, including a 1 up-regulated miRNA and 10 down-regulated miRNAs. Compared with normal esophageal samples, the ESC tissues showed up-regulated hsa-miR-126 and down-regulated hsa-miR-518b expression. CONCLUSION: hsa-miR-126 and hsa-miR-518b are differentially expressed in ESC, and they might play important roles in the carcinogenesis and progression of ESC.

[Expression and distribution of Nrf2 in several hepatocellular carcinoma cell lines].

AIM: The aim of this study was to investigate the expression and the distribution of Nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) in several human hepatocellular cancer cell lines (HepG2, Hep3B, SMMC-7721). METHODS: Flow cytometry was used to figure out the percentages of Nrf2-positive cells in these three cell lines. The localization of Nrf2 was estimated by the laser confocal microsopy and the expression levels of Nrf2 in hepatocellular cancer cell lines were detected by Western blot analysis. RESULTS: The percentages of Nrf2 positive cells in HepG2, Hep3B, and SMMC-7721 were 99.39%, 99.94%, and 99.98% through the flow cytometry. The laser confocal microsopy showed that Nrf2 mainly localized in the cytoplasm of HepG2 cells, distributed evenly in the cytoplasm and nucleus of Hep3B cells, and mainly localized in the nucleus of SMMC-7721 cells. Western blot analysis confirmed the result by the laser confocal microsopy. CONCLUSION: The data on the expression and localization of Nrf2 will be helpful for the following research on the role of Nrf2 in the drug resistance of hepatocellular carcinoma to chemotherapy.

[Effects of antidiabetic drug metformin on human breast carcinoma cells with different estrogen receptor expressing in vitro].

AIM: To research different effects of human breast carcinoma cells with different estrogen receptor expressing by antidiabetic drug metformin, and preliminary explore the possible underlying molecular mechanisms. METHODS: Cells were treated with metformin. Growth inhibition rates of the cells were measured by MTT assay. Cell cycle and apoptosis were detected by flow cytometery (FCM). Expressions of HIF-1α mRNA in the cells were measured by RT-PCR. RESULTS: After drug intervention, the cell proliferation were inhibited by metformin, and reinforced with the concentration and reaction time increase (P<0.05). The growth inhibition rates of MCF-7(ER(+);) breast carcinoma cell were higher than MDA-MB-231(ER(-);) breast carcinoma cell at each concentration group (P<0.05). The results of FCM prompted: MCF-7(ER(+);) breast carcinoma cell was arrested in G1 phase by metformin significantly in a dose-dependent increase(P<0.05). While for MDA-MB-231(ER(-);) breast carcinoma cell, only the 20 and 40 mmol/L groups had significant difference compared with the control group(P<0.05); and the percentage of G1 phase arresting were lower than MCF-7(ER(+);) breast carcinoma cell at the same concentration group(P<0.05). The effect of apoptosis for these two kinds of cells via metformin were not obvious(P<0.05). The expressions of HIF-1α mRNA detected by RT-PCR prompted: the expressions of HIF-1α mRNA for these two breast carcinoma cells were in a dose-dependent decrease(P<0.05). CONCLUSION: The effect of metformin for human breast carcinoma cell with estrogen receptor was better than the one without estrogen receptor. Maybe the molecular mechanism had a relationship with HIF-1α up-regulating.