Dianliang Zhang

Relationship between Matrix Metalloproteinase 2 and Lung Cancer Progression

AbstractBackground and objective: Matrix metalloproteinases (MMPs) play an important role in several steps of cancer development. MMP2 and MMP9 have previously been implicated in lymphatic and vascular invasion of lung cancer; however, the expression and prognostic significance of MMP2 and MMP9 is not fully clarified. This study was designed to investigate the significance of MMP2 and MMP9 in lung cancer tissue or serum, and their correlation with lung cancer prognosis. Methods: Immunohistochemical analysis was performed to determine MMP2 and MMP9 staining in human nonsmall cell lung cancer (NSCLC). Serum MMP2 and MMP9 protein levels in patients after surgery were measured using the ELISA method. The correlation between MMP2 and MMP9 serum levels and clinicopathological features of NSCLC were analyzed by survival analysis. We also performed reverse transcriptase (RT)-PCR assays to detect messenger RNA (mRNA) expression to further confirm the activity of MMP2 and MMP9 in human lung cancer. Results: Increased MMP2 immunostaining and MMP2 serum level correlated with advanced tumor stage and the presence of distant metastasis (Pearson’s χ2 test and ANOVA, p < 0.05). However, for MMP9, only serum level showed a correlation with advanced tumor stage. No significant correlation was observed between MMP2 or MMP9 immunostaining expression and tumor histologic features (Pearson’s χ2 test, p = 0.061 and p = 0.087, respectively). A high densitometry value of MMP2 and MMP9 PCR products (i.e. mRNA expression level) was related to poor differentiation grade, distant metastasis, and small cell carcinoma histologic type (ANOVA, p < 0.05). Conclusions: Our results suggest that MMP2 is a more sensitive predictor than MMP9 of lung cancer progression, metastasis, and survival. Serum MMP2 levels may be a valuable prognosis variable and could help to stratify lung cancer patients into low- and high-risk groups.

Nuclear Factor-κB Inhibition by Parthenolide Potentiates the Efficacy of Taxol in Non–Small Cell Lung Cancer In vitro and In vivo

In this study, we have examined the molecular events induced by parthenolide, a sesquiterpene lactone, and explored possible mechanisms of resistance and sensitization of tumor cells to Taxol. We showed that parthenolide could antagonize Taxol-mediated nuclear factor-κB (NF-κB) nuclear translocation and activation and Bcl-xl up-regulation by selectively targeting I-κB kinase activity. In A549 cells, inhibition of nuclear factor-κB by parthenolide resulted in activation of the mitochondrial death pathway to promote cytochrome c release and caspase 3 and 9 activation. In contrast, Taxol alone induced apoptosis via a pathway independent of mitochondria cytochrome c cascade. In addition, depletion of Bcl-xl rescued the apoptotic response to Taxol. Moreover, treatment with parthenolide increased the efficacy of the Taxol-induced inhibition of A549 tumor xenografts in mice. This study elucidated the cellular responses induced by parthenolide that decrease the threshold of mitochodria-dependent apoptosis in the treatment of non–small cell lung cancer cells. (Mol Cancer Res 2009;7(7):1139–49)

Potential biomarkers involving IKK/RelA signal in early stage non-small cell lung cancer

The clinical relevance of nuclear factor κB (NF‐κB) and its regulatory molecules on prognosis of patient with early stages of non‐small cell lung cancer (NSCLC), remains unclear. Therefore, we conducted biomarker analyses with survival in patients with stages I and II NSCLC. Tumor samples were collected from 88 patients with early‐stage NSCLC (stages I, II). A minimum follow‐up period of 5 years was required. RelA, phosphorylated IκB (pIκBα), pIKKα/β were detected by immunostaining. NF‐κB DNA binding activity was assessed by electrophoretic mobility shift assay. Association of clinical and pathologic variables (e.g. sex, age, pathologic stage) with relevant molecules was determined by Pearsons χ2 test or Fishers exact test. Survival analysis based on single expression of RelA, pIκBα, pIKKα/β as well as composite expressions were evaluated using Cox proportional hazards regression models, and log rank test followed Kaplan‐Meier estimates. RelA, pIκBα, pIKKα/β were observed as increased expression in NSCLC tissues compared with adjacent normal tissues and normal lung tissues. These molecules were associated with tumor‐node‐metastasis stages, T stages and histological status, respectively. Among the molecules analyzed, RelA and pIκBα‐positive were statistically significant predictors of patient death in the entire patient population adjusted by age, gender and smoking status; furthermore both RelA and pIκBα‐positive was the strongest prognostic indicators of poor prognosis by univariate and multivariate analyses. Borderline positive correlations were observed between RelA and pIκBα or pIKKα/β expression. In this cohort of early‐stage NSCLC patients, molecular markers, especially composite application of multiple biomarkers (both nuclear RelA and cytoplasmic pIκB‐α expression) that independently predict overall survival have been identified. (Cancer Sci 2008; 99: 582–589)

Combined Prognostic Value of Both RelA and IκB-α Expression in Human Non–Small Cell Lung Cancer

BackgroundWe sought to investigate the prognostic significance of nuclear factor (NF)-κB activity, especially nuclear RelA and IκB-α expression patterns, in non–small cell lung cancer (NSCLC).MethodsA total of 116 patients with pathologically confirmed stage I to II NSCLC were included. Immunohistochemical analysis and electrophoretic mobility shift assays of NF-κB were performed to determine RelA and phosphorylated IκB-α staining, and DNA binding activity of NF-κB in human NSCLC. Downstream genes, including VEGF and IL-8, were also assessed. The prognostic significance of a single expression of RelA, phosphorylated IκB-α, and b-composite expressions was evaluated by Cox proportional hazard regression models and by Kaplan-Meier survival analyses. Correlation between RelA/IκB-α expression status and clinicopathological features of NSCLC was also analyzed.ResultsNF-κB DNA binding activity, VEGF, and IL-8 showed correlation with nuclear RelA and cytoplasmic pIκB-α expression. Expression of nuclear RelA/NF-κB showed an increase in NSCLC tissue compared with adjacent normal tissue and normal lung tissue. There was a positive correlation between NF-κB activation (nuclear translocation of RelA) and tumor clinicopathological features such as tumor grade, including T stages, N stages, and tumor, node, metastasis system stages, smoking status, and age. Positive correlation was observed between nuclear RelA and cytoplasmic pIκB-α. Both nuclear RelA and cytoplasmic pIκB-α were associated with poor prognosis by univariate and multivariate analyses.ConclusionsNuclear RelA and cytoplasmic pIκB-α expression are associated with a poorer prognosis in NSCLC patients. In particular, composite application of these two biomarkers might be of greater value than application of a single marker to identify patients at high risk, even at an early clinical stage.

The altered tight junctions: an important gateway of bacterial translocation in cachexia patients with advanced gastric cancer.

UNLABELLED Tight junctions (TJs) are the structural basis for the intestinal epithelium barrier. Increased intestinal permeability caused by variations in TJ proteins may result in bacterial translocation (BT). There is increasing evidence that BT might contribute to the occurrence and development of cancer cachexia, but the details are not known. Aims, we undertook further investigations into the pathway of BT in cancer cachexia. RESULTS BT-positive patients had a higher level of claudins-2 (CL-2, P=0.035) and a lower level of occludin (P=0.038) and Zonula occluden-1 (P=0.01) than BT-negative patients. Moreover, the levels of IL-6, TNF-α, and IFN-γ in BT-positive cachexia patients were higher compared with BT-negative cachexia patients (P<0.001, P=0.01, P<0.001) and BT-positive noncachexia patients (P<0.001, P=0.025, P<0.001). In the BT-positive cachexia patients, the local concentration of IL-6, TNF-α, and IFN-γ, in the middle colic vein, was higher than in the peripheral venous (P=0.04, P=0.03, P=0.038). In addition, endotoxin was detected within the small intestinal wall, and the concentration of endotoxin decreased from the mucosal side to the serosal side gradually in BT-positive patients. This study suggests that the altered TJs could be an important gateway of BT in gastric cancer cachexia and local cytokines could play a more important role than systemic cytokines in the process.

Chemosensitization in non‐small cell lung cancer cells by IKK inhibitor occurs via NF‐κB and mitochondrial cytochrome c cascade

In this study, we demonstrated with mechanistic evidence that parthenolide, a sesquiterpene lactone, could antagonize paclitaxel‐mediated NF‐κB nuclear translocation and activation by selectively targeting I‐κB kinase (IKK) activity. We also found that parthenolide could target IKK activity and then inhibit NF‐κB; this promoted cytochrome c release and activation of caspases 3 and 9. Inhibition of caspase activity blocked the activation of caspase cascade, implying that the observed synergy was related to caspases 3 and 9 activation of parthenolide. In contrast, paclitaxel individually induced apoptosis via a pathway independent of the mitochondrial cytochrome c cascade. Finally, exposure to parthenolide resulted in the inhibition of several NF‐κB transcript anti‐apoptotic proteins such as c‐IAP1 and Bcl‐xl. These data strengthen the rationale for using parthenolide to decrease the apoptotic threshold via caspase‐dependent processes for treatment of non‐small cell lung cancer with paclitaxel chemoresistance.

Interleukin-10 Gene Polymorphisms Influence Susceptibility to Cachexia in Patients with Low-Third Gastric Cancer in a Chinese Population

AbstractBackground and Objective: Interleukin (IL)-10 is a pleiotropic cytokine that can both stimulate and suppress the immune response. Previous studies have reported that IL-10 production was significantly elevated in cachectic patients, and it has been confirmed that polymorphisms of the IL10 gene could influence its expression. Therefore, we designed this study to investigate whether polymorphisms of the IL10 gene were associated with cachexia in patients with low-third gastric cancer in a Chinese population. Methods: 190 patients with low-third gastric cancer were included in this study. The serum levels of IL-10 were measured by radioimmunoassay. The single nucleotide polymorphisms (SNPs) at positions -1082A/G, -819T/C, and -592A/C in the IL10 gene promoter were analyzed using polymerase chain reaction (PCR) restriction fragment length polymorphism (PCR-RFLP). Results: The serum levels of IL-10 were significantly higher in patients with cachexia than in those without (Z = −10.66, p<0.001). Single SNP analysis showed that the frequency of the IL10 -1082G allele was increased in patients with cachexia (p = 0.02). The -1082AG and -819CC genotypes were observed to be associated with an increased risk of cachexia. In a logistic regression analysis adjusted for actual weight and carcinoma stage, the -1082AG genotype was associated with an odds ratio (OR) of 2.45 (95% CI 1.21, 4.96; p = 0.01), and the -819CC genotype was associated with an OR of 3.70 (95% CI 1.20, 11.39; p = 0.02) for cachexia. Furthermore, haplotype analysis of the -1082A/G, -819T/C, and -592A/C SNPs revealed that at least five haplotypes (ATA, ACC, GCC, ACA, and ATC) were present in this Chinese population, and the -1082G/-819C/-592C (GCC) haplotype was associated with a significantly increased risk of cachexia as compared with the ATA haplotype (OR = 2.42; 95% CI 1.17, 5.00; p = 0.02). Conclusion: Our results indicate that genetic polymorphisms of IL-10 may influence susceptibility to cachexia in patients with low-third gastric cancer in this Chinese population.

Serum biomarker screening for the diagnosis of early gastric cancer using SELDI-TOF-MS

In this study, we performed a proteomic analysis of sera from stage I gastric cancer patients using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and established a diagnostic model for the early diagnosis of stage I gastric cancer. Serum samples from 169 gastric cancer patients and 83 age- and gender-matched healthy individuals were analyzed by SELDI-TOF-MS ProteinChip array technology. The SELDI-TOF-MS spectral data were analyzed using the Biomarker Wizard™ and Biomarker Patterns™ software to find differential proteins and develop a classification tree for gastric cancer. A total of 34 mass peaks were identified. Six peaks at a mass-to-charge ratio (m/z) of 2873, 3163, 4526, 5762, 6121 and 7778 were used to construct the diagnostic model. The model effectively distinguished gastric cancer samples from control samples, achieving a sensitivity and specificity of 93.49 and 91.57%, respectively. In addition, we identified 3 of the 6 protein peaks at 2873, 6121 and 7778 m/z, which distinguished between stage I and stage II/III/IV gastric cancer. The model had an accuracy of 88.89% for the identification of stage I gastric cancer. In conclusion, the diagnostic model for the detection of serum proteins by SELDI-TOF-MS ProteinChip array technology correctly distinguishes gastric cancer from healthy samples, and has the ability to screen and distinguish between early gastric cancer from advanced gastric cancer.

TLR and MBL Gene Polymorphisms in Severe Acute Pancreatitis

AbstractBackground and objective: Mannose-binding lectin (MBL) and toll-like receptor (TLR)-4 gene polymorphisms have been implicated in inflammatory episodes in a number of studies. In view of the inflammatory nature of acute pancreatitis, we aimed to determine the predictive value of two point mutations in the promoter region at position −550 (H/L variants) and −221 (X/Y variants) of the MBL2 gene, and the Asp299Gly and 119C>A polymorphisms of the TLR4 gene on the occurrence of severe acute pancreatitis (SAP). Methods: The study included 132 patients with SAP, 106 with mild acute pancreatitis (MAP), and 121 healthy volunteers. Genotypes were determined using restriction fragment length polymorphism analysis of PCR products and by allele-specific PCR. Results: No significant difference in genotype frequency was noted between the patients with acute pancreatitis and controls for any of the gene loci studied. The distributions of the HY/HY, HY/LY, LY/LY, and LY/LX genotypes of MBL2 gene promoter and 119C>A genotype of the TLR4 gene were similar in patients with mild or severe acute pancreatitis. HY/LX genotype frequency was significantly higher in patients with SAP compared with MAP (26% vs 14%; p = 0.028). Conclusion: Results indicate that the MBL2 HY/LX genotype plays an important role in the determination of disease severity to acute pancreatitis.

Bacterial translocation contributes to cachexia and its possible pathway in patients with colon cancer.

Goals and Background: There is increasing evidence that bacterial translocation (BT) might contribute to the occurrence and development of cancer cachexia, but the detailed mechanism remains unknown. Thus, we undertook further investigations into the association of BT with cancer cachexia and the possible pathway. Study: The colon cancer patients enrolled in this study were divided into cachectic and noncachectic. BT was analyzed by polymerase chain reaction and bacterial culture. Intestinal epithelial T-cell subsets and NK cells were evaluated using flow cytometry. Western blotting and immunofluorescence were used to check tight junction (TJ) proteins in intestinal epithelium. Fluorescence in situ hybridization and immunohistochemistry were used to detect the translocated bacteria and endotoxin. Results: Compared with noncachectic patients, cachectic patients had a significantly higher BT ratio (P<0.001). We observed the translocated bacteria in the intestinal mucus layer associated with lower levels of T-cell subsets and NK cells in the intestinal epithelium in BT-positive patients (P<0.05). Endotoxin was detected within the small intestinal wall and the concentration of endotoxin decreased from the mucosal side to serosal side gradually in these patients. These were associated with an altered composition of TJs. Conclusions: This study suggests that BT may contribute to colon cancer in cachectic patients, and TJ could be the gateway to the possible pathway of BT.