Gene Expression Levels In Patients Research Articles

Assessment of suitable reference genes for RT\u2013qPCR studies in chronic rhinosinusitis

Reverse transcription–quantitative polymerase chain reaction is a valuable and reliable method for gene quantification. Target gene expression is usually quantified by normalization using reference genes (RGs), and accurate normalization is critical for producing reliable data. However, stable RGs in nasal polyps and sinonasal tissues from patients with chronic rhinosinusitis (CRS) have not been well investigated. Here, we used a two-stage study design to identify stable RGs. We assessed the stability of 15 commonly used candidate RGs using five programs—geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. Ribosomal protein lateral stalk subunit P1 (RPLP1) and ribosomal protein lateral stalk subunit P0 (RPLP0) were the two most stable RGs in the first stage of the study, and these results were validated in the second stage. The commonly used RGs β-actin (ACTB) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were unstable according to all of the algorithms used. The findings were further validated via relative quantification of IL-5, CCL11, IFN-γ, and IL-17A using the stable and unstable RGs. The relative expression levels varied greatly according to normalization with the selected RGs. Appropriate selection of stable RGs will allow more accurate determination of target gene expression levels in patients with CRS.

Altered Expression of TXNIP in the peripheral leukocytes of patients with coronary atherosclerotic heart disease.

Background:Coronary atherosclerotic heart disease (CAD) is mainly caused by atherosclerosis, an inflammatory disease characterized by plaque formation in arteries. Reactive oxygen species caused structural damage and dysfunction of arterial endothelial cells. Thioredoxin-interacting protein (TXNIP) is the endogenous inhibitor and regulator of thioredoxin, a major cellular antioxidant and antiapoptotic system. In order to explore the role of TXNIP in the occurrence and development of CAD, we detected the TXNIP expression and discussed its molecular mechanisms in CAD.Methods:The mRNA levels of TXNIP gene in peripheral leucocytes were detected in CAD and healthy controls (CTR) by quantitative real-time polymerase chain reaction. And TXNIP proteins were detected by western blotting.Results:TXNIP gene expression levels in patients with unstable angina pectoris (UAP, n = 96) were significantly increased compared with those of CTR (n = 192, P < .05). However, the situation is different in acute myocardial infarction (n = 96, P > .05). Logistic regression analysis showed that TXNIP levels were significantly positive correlated with UAP (OR = 1.728, P < .05).Conclusions:TXNIP gene expression in the peripheral leucocytes was increased in patients with UAP, indicating that TXNIP in circulating leucocytes may be involved in the pathogenesis of UAP.

Increased platelet alpha2B-adrenergic receptor gene expression in well-controlled hypertensives: the effect of arterial stiffness

Catecholamines play a major role in atherothrombotic mechanisms in essential hypertension. Alpha2B-adrenergic receptors (α2B-ARs) are implicated in the pathophysiology of platelet aggregation. In this study, we evaluated platelet α2B-AR gene expression levels in patients with well-controlled essential hypertension compared with normal individuals and investigated their association with increased arterial stiffness. Fifty-nine patients with well-controlled essential hypertension (34 men, mean age 65 ± 9 years) and 26 normotensives (19 men, mean age 64 ± 8 years) were included in the study. For each patient, carotid-femoral pulse wave velocity (PWV) and carotid-radial PWV were evaluated. In addition, blood samples were obtained and platelets were isolated. The α2B-AR gene expression levels in platelets were examined by real-time polymerase chain reaction for each participant. Well-controlled hypertensive patients showed significantly higher gene expression levels of α2B-Rs in platelets compared with normotensives (34.7 ± 29.5 vs 17.6 ± 12.5, respectively, P = .005). Interestingly, we found that carotid-femoral PWV and carotid-radial PWV were positively correlated with platelet α2B-R gene expression levels (r = 0.59, P < .001, and r = 0.39, P = .002, respectively).Platelet α2B-R gene expression levels are increased in patients with well-controlled essential hypertension compared with normotensives and are correlated with increased PWV in those patients. Our data indicate an association of arterial stiffness and platelet α2B-Rs gene expression and indicate the need for further research.

Heavy alcohol drinking downregulates ALDH2 gene expression but heavy smoking up-regulates SOD2 gene expression in head and neck squamous cell carcinoma

BackgroundThis study aims to determine the relationship between expression levels of ALDH2 and SOD2 genes and clinical parameters such as alcohol drinking, tobacco smoking, primary site of HNSCC, and human papilloma virus (HPV) state.MethodsGene expression data were obtained from gene expression omnibus (GEO accession number: GSE65858). Clinical data (N = 270) including survival result, gender, age, TNM stage, primary site of HNSCC, HPV status, alcohol drinking, and tobacco smoking habit were analyzed according to gene expression pattern.ResultsALDH2 gene was expressed in low levels in patients with heavy alcohol consumption. It was expressed in high (p = 0.01) levels in patients with no or light alcohol consumption. ALDH2 gene was also expressed in low levels in patients with oral cavity cancers or hypopharynx cancers. However, ALDH2 gene was expressed in high (p = 0.03) levels in patients with oropharyngeal cancers or laryngeal cancers. HPV-positive patients were found to have high (p = 0.02) expression levels of ALDH2. SOD2 gene was expressed in high (p = 0.005) levels in patients who had greater mean pack-year of tobacco smoking. Based on log rank test, the group of patients with high expression of ALDH2 showed better (p = 0.002) clinical results than those with low expression of ALDH2. Difference of survival results between ALDH2 high-expressed group and ALDH2 low-expressed group was validated in another cohort (GSE39368, N = 138).ConclusionsHeavy alcohol drinking downregulates ALDH2 gene expression level. Heavy smoking up-regulates SOD2 gene expression level in patients with head and neck squamous cell carcinoma. The group of patients with low expression levels of ALDH2 showed significantly poorer survival results compared to those with high expression levels of ALDH2.

P2579Differential microRNA gene expression levels in patients with acute myocarditis

P2579Differential microRNA gene expression levels in patients with acute myocarditis

Frequency of MCP-1 (rs1024611) and CCR2 (rs1799864) gene polymorphisms and its effect on gene expression level in patients with AgP

ObjectiveTo identify the genetic risk markers of aggressive periodontitis (AgP), researchers focus on genetic components that regulate the immune response. Therefore the purpose of this study was to investigate genetic impact of monocyte chemoattractant protein (MCP)-1–2518A/G and CC chemokine receptor 2 (CCR2) −190G/A gene polymorphisms on AgP susceptibility and the effect of this polymorphism on MCP-1 gene expression in patients with AgP. Material and methodsA total of 215 subjects, 108 AgP and 107 periodontally healthy (H) were recruited in this cross-sectional study (NCT02817568). Gene polymorphisms of MCP-1–2518A/G and CCR2–190G/A were analyzed by a standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. MCP-1 messenger (m) RNA expression was measured using quantitative real-time (RT)-PCR in peripheral blood leukocytes from 26 AgP and 16H controls. Threshold cycles (Ct) values were obtained from the RT-PCR analysis based on SYBR Green detection and data was normalized via ΔCt. ResultsThere were no differences between AgP and H groups with regard to MCP-1 and CCR2 genotype distribution and allele frequencies (p>0.05). In contrast, the MCP-1 mRNA expression levels were higher in homozygous “AA” control subjects than having G+ genotype and AA homozygous AgP patients. ConclusionsIt can be concluded that MCP-1 and CCR2 polymorphisms are not associated with AgP in Turkish population. Although in AgP patients, there was AA genotype with MCP-1 mRNA expression it can be speculated that gene expression levels in peripheral blood may not reflect the cytokine/chemokine levels of local tissues.

Testosterone treatment increases androgen receptor and aromatase gene expression in myotubes from patients with PCOS and controls, but does not induce insulin resistance

Polycystic ovary syndrome (PCOS) is associated with insulin resistance and increased risk of type 2 diabetes. Skeletal muscle is the major site of insulin mediated glucose disposal and the skeletal muscle tissue is capable to synthesize, convert and degrade androgens. Insulin sensitivity is conserved in cultured myotubes (in vitro) from patients with PCOS, but the effect of testosterone on this insulin sensitivity is unknown. We investigated the effect of 7days testosterone treatment (100nmol/l) on glucose transport and gene expression levels of hormone receptors and enzymes involved in the synthesis and conversion of testosterone (HSD17B1, HSD17B2, CYP19A1, SRD5A1-2, AR, ER-α, HSD17B6 and AKR1-3) in myotubes from ten patients with PCOS and ten matched controls.Testosterone treatment significantly increased aromatase and androgen receptor gene expression levels in patients and controls. Glucose transport in myotubes was comparable in patients with PCOS vs. controls and was unchanged by testosterone treatment (p=0.21 PCOS vs. controls). These results suggest that testosterone treatment of myotubes increases the aromatase and androgen receptor gene expression without affecting insulin sensitivity and if testosterone is implicated in muscular insulin resistance in PCOS, this is by and indirect mechanism.

The in vitro effect of oxidized LDL and PHA on proliferation and gene expression of regulatory T cells in patients with atherosclerosis.

Atherosclerosis is a chronic inflammatory condition that affects the arterial wall. Oxidized low-density lipoprotein (ox-LDL) seems to have an important role in atherosclerotic plaque formation.This study was performed to investigate the effects of ox-LDL as well as PHA on proliferation and gene expression of peripheral blood mononuclear cells (PBMCs) in patients with atherosclerosis compared to healthy controls. Proliferation of PBMCs was assessed by BrdU assay, while gene expression was assessed by real-time PCR. Both PHA and ox-LDL significantly induced proliferation of PBMCs of patients and controls. PBMCs from controls showed significantly higher proliferation when stimulated with ox-LDL compared to patients. Expression of TGF-β was significantly lower in PBMCs from patients compared to healthy controls (p<0.001). Following simulation with PHA, TGF-β and Foxp3 gene expression levels in patients and controls were significantly decreased (p<0.001). Expression of Foxp3 in PBMCs treated with ox-LDL was significantly decreased in patients and controls.Decreased expression of TGF-β and Foxp3 genes after ox-LDL stimulation may be due to more sensitivity of Treg cells than effector T cells to ox-LDL. Presence of ox-LDL within atheroma could be associated with the diminished population of Treg cells in the atherosclerotic patients.

S100A9 and S100A8 gene expression levels in patients with psoriasis

S100A9 and S100A8 gene expression levels in patients with psoriasis

The Expression of GATA-3 and FoxP3 Genes and the Frequency of Tregs Are Correlated with Gvhd Following Allogeneic HSCT

AbstractAbstract 5181 ObjectiveHematopoietic stem cell transplantation (HSCT) is a common therapeutic option for leukemia, but its use is limited by mortality due to graft-versus-host-disease (GVHD). Naturally occurring regulatory T cells (Tregs) are mediators of immunologic tolerance that attenuate GVHD in experimental models. Tregs were first defined by a CD4+CD25+ phenotype, but subsequent studies identified forkhead box protein 3 (FoxP3) as a highly specific marker in both mouse and human T cells with regulatory function. GATA-3 is a member of the GATA family of transcription factors, and is sufficient to induce a Th2 phenotype. GATA-3 could act as an inhibitor of FoxP3 expression in early T cell differentiation was found in transgenic mice model recently. But whether GATA-3 expression could correlate with GVHD incidence, or whether GATA-3 could influence Tregs recovery and contribute to the development of GVHD are still not clear. So we investigated the frequency of Tregs in the early period after HSCT and at the time of GVHD onset, and quantitatively measured the expression of GATA-3 and FoxP3 at GVHD onset for analyzing the correlation to the development of GVHD. MethodsSixty-seven leukemia patients underwent allogenic (allo-) HSCT were enrolled in this investigation, including 45 males and 22 females (median age: 32.3 years, range 21≂f45). Fourteen healthy individuals (7 males and 7 females, median age: 30.3 years, range 26≂f40) served as controls. Frequencies of T lymphocyte subsets were determined by flow cytometry. The expression levels of human FoxP3 and GATA-3 genes were determined by real-time quantitative PCR with SYBR Green I technique. ResultsThe frequencies of CD4+ CD25+ T cells were lower at week 4 post-HSCT, but significant higher after week 4 and lower again after week 8 (P=0.000). The frequencies of FoxP3+ T cells were also lower at week 4 and remained low levels within 12 weeks after HSCT. As early as 4 to 8 weeks after HSCT, the frequencies of CD4+ FoxP3+ T cells were lower than that of week 4 (P=0.048), but the frequencies of CD25+ FoxP3+ T cells were higher than that of week 4 (P=0.022).Frequencies of CD4+ CD25+ cells in patients at GVHD onset were similar in patients without GVHD (P=0.262) or healthy individuals (P=0.166). But the frequencies of FoxP3+ cells, FoxP3+ CD25+ cells and FoxP3+ CD4+ cells in patients at GVHD onset were significantly lower than that in patients without GVHD (P=0.000, 0.004, and 0.001). The FoxP3 gene expression levels in patients at GVHD onset were significantly lower than that in patients without GVHD (P=0.044), but the GATA-3 gene expression levels were significantly higher in patients at GVHD onset compared to patients without GVHD (P=0.005). The FoxP3 gene expression levels in patients at aGVHD onset were significantly higher than that in patients at cGVHD onset (P=0.038), but the GATA-3 gene expression levels were significantly lower in patients at aGVHD onset compared to patients at cGVHD onset (P=0.004).A highly significant correlation between FoxP3 and GATA-3 genes expression levels existed in healthy individuals (Spearman correlation coefficient r=-0.893, P=0.007). Correlations between FoxP3 and GATA-3 genes expression levels were also significant in patients at aGVHD onset (Pearson correlation coefficient r=0.749, P=0.020) or cGVHD onset (Pearson correlation coefficient r=0.538, P=0.071) or in GVHD (Spearman correlation coefficient r=0.370, P=0.090). But there is no significant correlation between FoxP3 and GATA-3 genes expression levels in patients without GVHD (Pearson correlation coefficient r=-0.141, P=0.821). ConclusionsGATA-3 and FoxP3 genes expression and the frequency of Tregs are correlated with GVHD following allogeneic HSCT. GATA-3 can influence the expression of FoxP3 in the development of GVHD.Supported by National Natural Science Foundation of China(30971300), Science and Technology Planning Project of Guangdong Province of China (2009A030200007) and China Postdoctoral Science Foundation (200902332, 20080440776). Disclosures:No relevant conflicts of interest to declare.

P122 Protein Enhances Intracellular Calcium Increase to Acetylcholine

Phospholipase C-δ1 activity is enhanced in patients with coronary artery spasm, and a p122 protein was recently cloned to potentiate phospholipase C-δ1 activity. To investigate the role of p122 in enhanced vasomotility, we examined p122 expression in the cultured skin fibroblasts obtained from patients with and without coronary spasm, intracellular Ca(2+) concentration ([Ca(2+)]i) [corrected] at baseline and after stimulation with acetylcholine in the cells transfected with p122, and promoter in genomic DNA. [corrected] p122 protein and gene expression levels in patients with coronary spasm (n=11) were enhanced compared with levels in control subjects (n=9) (P<0.01 for both). [Ca(2+)](i) at baseline and the peak increase in [Ca(2+)](i) in response to acetylcholine were both 2 times higher in cells transfected with p122 than in those without p122. Conversely, knockdown of p122 resulted in diminished [Ca(2+)](i) response. In the p122 promoter analysis, the -228G/A and -1466C/T variants revealed the increase in luciferase activity. Although the -1466C/T variant was similar between 144 patients with coronary spasm and 148 controls, the -228G/A variant was more frequent in male patients than in male controls (P<0.05). The p122 protein is upregulated in patients with coronary spasm, causing increased [Ca(2+)](i) to acetylcholine, and thereby seems to be related to enhanced coronary vasomotility.

The investigation of CD36 scavenger receptor and MSR1 scavenger receptor gene expression level in patients with atherosclerosis

The investigation of CD36 scavenger receptor and MSR1 scavenger receptor gene expression level in patients with atherosclerosis

Human Multidrug Resistance-1 Gene Expression Levels in Graves-Basedow Disease

Multidrug resistance 1 gene is responsible for the resistance of a large variety of drugs in human cells. We tried to evaluate this in the present study in thyroid stimulant hormone receptor antibody positive subjects. In study enrolled 23 female and 10 male subjects. Hyperthyroid subjects were treated with PTU and remission was assured between 6-12 months. Blood samples were collected before the start of this treatment. Permission for this study was taken from the patients and the local ethical committee. Serum F-T3, F-T4 levels in Graves subjects were markedly high, whereas TSH levels were markedly low than normal range. We also found that with increased age of the Graves' subjects, MDR-1 gene expression decreased. There was also a direct correlation between blood MDR-1 gene expression and TSH-R Ab levels in patients with Graves's disease. We observed that the duration of being euthyroid was lengthened with the elevation of MDR-1 gene expression. There was a direct correlation between blood MDR-1 gene expression levels and ultrasonografic size of thyroid gland. As a result, raised blood MDR-1 gene expression levels in patients with Graves-Basedow disease may be associated with the activity of the disease and the resistance to its treatment. The more blood MDR-1 expression increases the more the duration of being euthyroid increases.

Molecular profiling in patients (pts) with upper gastrointestinal (UGI) cancers correlated with clinical outcome

e22042 Background: Recent efforts have expanded our understanding of the molecular pathogenesis of UGI cancers and how oncogenes and tumor suppressor genes play a role in both carcinogenic and metastatic processes. These markers may serve as prognostic and/or predictive factors for recurrence following resection and resistance to radiotherapy and chemotherapy. Gene expression levels of thymidylate synthase (TS), thymidylate phosphorylase (TP), epidermal growth factor receptor (EGFR) and excision repair cross complementing (ERCC-1) have been shown to be associated with outcomes in lung, gastric and colon cancer. We evaluated TS, ERCC-1, EGFR and TP gene expression levels in patients with UGI cancers. Methods: This was a retrospective study of 80 pts with UGI cancers evaluated at USC who underwent molecular profiling. The primary objective was to determine a correlation between TS, TP, ERCC1 and EGFR and correlate with clinical outcome. Characteristics of these 80 pts: (16 females, 64 males ); median age 61 years (range 34–85); tumor types evaluated - 32 (40%) esophageal, 24(30%) gastric, 24(30%) GE junction; stages- 53 % IV, 21% III, 12% II, 2% I were evaluated for intratumoral gene expression of TS, TP, ERCC-1, &amp; EGFR by real time quantitative PCR using Taqman technology from microdisected paraffin-embedded tumor sections. Results: High TS expression was associated with shorter OS (12.8 months vs. 23.7 months p=0.036). A significant correlation was found between TP &amp; ERCC-1 (p=0.0078, r=0.37); TP &amp; TS (p=0.0128, r=0.35); TP &amp; EGFR (p=0.0065, r=0.39); TS &amp; ERCC-1 (p=0.0004, r=0.39); ERCC-1 &amp; EGFR (p=0.025, r=0.30). No statistically significant relationship was found between TS &amp; EGFR (p=0.06,r=0.25). There was no correlation of ERCC-1, TP, &amp; EGFR with OS that reached statistical significance. Conclusions: TS mRNA levels were shown to be associated with OS in UGI tumors, consistent with data reported in colon cancer. TS gene expression was significantly associated with expression levels of ERCC-1. In addition, ERCC1 was associated with EGFR. These data show for the first time that molecular pathways of cytotoxic agents are linked to the EGFR pathway suggesting that sensitivity to fluoropyrimidines, oxaliplatin, and EGFR inhibitors may be associated. No significant financial relationships to disclose.

Clinicopathological significance of circadian rhythm‐related gene expression levels in patients with epithelial ovarian cancer

Recent studies implicate circadian genes in the regulation of cell cycle, apoptosis, and cell proliferation at a molecular level. These genesey affect cancer incidence, prognosis, and chemosensitivity. In this study, we measured the expression levels of clock genes and correlated their expression levels with clinicopathological parameters in epithelial ovarian cancer. The expression levels of core clock genes, per1, per2, per3, cry1, cry2, Bmal1, clock, and CKIepsilon were quantified by real-time quantitative Reverse transcription-polymerase chain reaction in 83 ovarian cancer tissues and 11 normal ovarian tissues. The expression levels of per1, per2, cry2, clock, CKIepsilon in ovarian cancers were significantly lower than those in normal ovaries. In contrast, cry1 expression was highest among the eight examined clock genes, followed by per3 and Bmal1. Cry1 expression was much higher in cancer than that in normal ovaries. Localized circadian gene expression was determined in cancer cells by in situ hybridization analysis. Cry1 expression was significantly reduced in mucinous and grade 3 tumors. Bmal1 expression was also significantly reduced in mucinous adenocarcinomas as compared to other histologies. In a multivariate analysis, the combination of low cry1 expression and low Bmal1 expression was an independent prognostic factor, as well as stage and histological subtype. The antiphase expression of cry1 and Bmal1 may be preserved in ovarian cancers. The combination of cry1 and Bmal1 expression might become a possible prognostic marker in epithelial ovarian cancer.

Treatment With Topical Steroids Downregulates IL-5, Eotaxin-1/CCL11, and Eotaxin-3/CCL26 Gene Expression in Eosinophilic Esophagitis

Our aim was to evaluate the changes induced by topical steroid treatment to the esophageal epithelial inflammatory eosinophilic and T-cell infiltrate and to IL-5, eotaxin-1/CCL11, and eotaxin-3/CCL26 esophageal gene expression levels in patients with eosinophilic esophagitis (EE). Esophageal biopsies were taken from eight adult patients at the moment of diagnosis and after 3-month treatment with fluticasone propionate. Eosinophils, CD8, and CD4 T cells were examined by immunohistochemistry. IL-5, eotaxin-1/CCL11, and eotaxin-3/CCL26 gene expression levels were measured by real-time PCR. Eight control samples were also analyzed. A significant decrease in the eosinophil infiltrate and in CD8(+) T-cell density was observed in the esophageal epithelium from the patients upon steroid treatment. IL-5 was not detected in control samples, and expression levels were variably downregulated after treatment in six of the patients. Gene expression of eotaxin-1/CCL11 showed relevant downregulation in four cases and a modest twofold decrease in three of the patients studied. Mean CCL11 expression values upon steroid treatment were similar to control samples (19.4 +/- 28.6 vs 8.42 +/- 5, P= 0.7). Eotaxin-3/CCL26 gene expression levels were significantly increased in EE. Although they were significantly downregulated upon steroid treatment, control expression levels were not reached in any of the cases analyzed (580.9 +/- 943.9 vs 1.45 +/- 1.0, P= 0.001). Our results confirm that eotaxin-3/CCL26 is significantly increased in EE esophageal samples. However, the individual analysis of IL-5, CCL11, and CCL26 expression data suggests that several cytokines and chemokines could participate in the physiopathology of EE in humans.

Genomic regulation of CTLA4 and Multiple Sclerosis

The cytotoxic T lymphocyte antigen 4 gene (CTLA4) is a critical regulator of T-cell activation and it is an important therapeutic target for cancer and autoimmune diseases. Here, we analyzed the genomic regulation of CTLA4 gene expression in order to identify single nucleotide polymorphisms (SNPs) that affect its expression and splicing, and to assess their association with Multiple Sclerosis (MS). We analyzed 152 healthy subjects and 146 patients with MS, of which 52 controls and 51 patients were used for gene expression analysis. We genotyped 17 SNPs in the CTLA4 gene using the SNaPshotTM Multiplex Kit, and in addition gene expression of the soluble (sCTLA4) and full length (flCTLA4) isoforms was quantified by real-time PCR, while protein levels of sCTLA4 were measured by ELISA. We found that the SNPs at − 1577, + 6230, + 10242, + 10717 and + 12310 influence CTLA4 expression and the combination of the − 1577 GG and + 6230 GG genotypes provokes the strongest decrease in CTLA4 gene expression. We found that the SNP at − 658 only acted as a regulatory SNP in patients with MS, suggesting the existence of epigenetic changes due to this disease. We also identified a decrease in CTLA4 gene expression levels in patients receiving chemotherapy, although no association was observed between MS and any of the polymorphisms studied. In conclusion, we have identified several SNPs in the CTLA4 gene and studied their influence on its genetic regulation. The involvement of CTLA4 in the pathogenesis of MS may be subtle and related to the functional changes in its pathway rather than predisposing genetic polymorphisms.

Significance of thymidylate synthase gene expression level in patients with adenocarcinoma of the lung

Thymidylate synthase (TS) catalyzes the methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) and is a key enzyme for DNA synthesis. High expression of TS is thought to be associated with poor prognosis in some kinds of cancers. However, this association has not been clarified for nonsmall cell lung carcinoma. In the current study, the authors investigated the clinicopathologic significance of TS mRNA levels and the correlation with cellular proliferation in patients with lung adenocarcinoma. The expression levels of TS mRNA were measured in 47 lung adenocarcinoma tissues using the Taq-Man real-time reverse transcriptase-polymerase chain reaction (RT-PCR) method and examined the clinicopathologic significance of TS expression. To clarify the correlation between TS expression level and cell proliferation, the Ki-67 labeling index was also examined using immunohistochemical staining. A positive correlation was observed between the expression levels of TS mRNA and stage of disease, lymph node metastasis, or tumor differentiation (P = .015). The higher expression group of TS mRNA showed a significantly poorer prognosis than the group with lower expression (P = .042). Moreover, there was a strong correlation noted between the expression levels of TS mRNA and the Ki-67 labeling index (P = .009). The results of the current study demonstrated that TS may be associated with stage of disease, lymph node metastasis, tumor differentiation, prognosis, and tumor cell proliferation. These results suggest that the expression levels of TS mRNA may be useful for predicting the malignant potential in patients with lung adenocarcinoma.

Tumor Cyclooxygenase-2 Gene Suppresses Local Immune Responses in Patients with Hepatocellular Carcinoma

In several neoplastic diseases including hepatocellular carcinoma (HCC) immunosuppression is correlated with disease stage, progression and outcome. Moreover, recent studies have demonstrated that cyclooxygenase-2 (COX-2) enhances tumor growth in HCCs. The present study analyzed the correlation between local immune responses and COX-2 gene expression levels in patients with primary HCCs. Fresh tissues were obtained from 59 patients who underwent resection of an HCC. The COX-2 gene expression levels were quantified by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and compared with the CD8+ T cell densities detected by immunohistochemistry. COX-2 gene expression was detected in 35 of the 59 tumors. The CD8+ T cell density in COX-2-expressing tumors (6.1 cells/high-power field (HPF), x200 magnification) was suppressed compared with that in non-COX-2-expressing tumors (13.6 cells/HPF, P = 0.009). Tumor COX-2 gene expression was associated with a poorer disease-free survival rate. Elevation of the tumor COX-2 level is correlated with the suppression of local immune responses in HCCs, suggesting that COX-2 plays a role in early tumor recurrence in the residual liver in patients after HCC resection.

Expression of the Multidrug Resistance-associated Protein (mrp ) Gene in Chronic Lymphocytic Leukemia

In order to define more accurately the role of multidrug resistance (MDR)-related protein (mrp ) gene in chronic lymphocytic leukemia (CLL), we addressed the question of its expression pattern in isolated peripheral blood B lymphocytes in seven healthy donors and 28 patients with CLL, with respect to some laboratory and clinical parameters of the disease. For this purpose, we used a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), based on the coamplification of an internal standard not homologous to the DNA target to quantify the mrp transcription level in each studied sample. We report that the level of constitutive mrp gene's expression in peripheral blood lymphocytes is higher in CLL patients than in healthy controls. We found increased mrp gene expression levels in patients with higher white blood cells (WBC) and lymphocytes' counts as well as in more advanced disease stages according to Rai or Binet scale. Finally, mrp gene's expression was higher in patients with progressive CLL, especially in cases refractory to chemotherapy salvage. The results of the present study suggest that expression of mrp gene might be relevant in the pathogenesis of the MDR phenotype in CLL.