High EGFR copy number predicts benefits from tyrosine kinase inhibitor treatment for non-small cell lung cancer patients with wild-type EGFR.

BACKGROUND: This study was designed to determine whether advanced non-small-cell lung cancer (NSCLC) patients with high copy number of epidermal growth factor receptor (EGFR) can benefit from treatment with EGFR-tyrosine kinase inhibitors (TKIs). METHODS: EGFR gene copy number was assessed by fluorescence in situ hybridization (FISH) and EGFR mutations was tested using Luminex xTAG technology in 502 TKI-treated NSCLC patients. The association between both biomarkers and clinical benefit from EGFR-TKI were analyzed. RESULTS: EGFR FISH+and EGFR mutations were significantly associated with higher response rates (37.2% and 43.7%, respectively), superior progression-free survival (PFS) (FISH+, 11.2 months; hazard ratio [HR], 0.51; 95% CI, 0.42 to 0.62; p<0.001; mutation+, 11.7 months; HR, 0.37; 95% CI, 0.31 to 0.45; p<0.001) and overall survival (OS) (FISH+, 30.2 months; HR, 0.51; 95% CI, 0.40 to 0.65; p<0.001; mutation+, 30.2 months; HR, 0.45; 95% CI, 0.36 to 0.58; p<0.001). In patients with wild-type EGFR, EGFR FISH+correlated with longer PFS than EGFR FISH- status (4.4 months vs. 2.0 months; HR, 0.56; 95% CI, 0.41 to 0.75; p<0.001), so did amplification (5.0 months vs. 2.0 months; HR, 0.43; 95% CI, 0.24 to 0.76; p=0.003). However, FISH+had no association with improved PFS in EGFR-mutated patients (HR, 0.77; 95% CI, 0.57 to 1.03; p=0.076). CONCLUSIONS: A combined analysis of EGFR FISH and mutation is an effective predictor of EGFR-TKI therapy. Specifically, a high EGFR copy number may predict benefit from TKIs treatment for NSCLC patients with wild-type EGFR.

Reduced expression of Krüppel-like factor 17 is related to tumor growth and poor prognosis in lung adenocarcinoma.

Krüppel-like factor 17 (KLF17), a new member of the Krüppel-like factors (KLFs), has been reported to be a negative regulator of epithelial-mesenchymal transition (EMT) and metastasis in breast cancer. However, the biological role and clinical significance of KLF17 in lung adenocarcinoma has been less clear. In the present study, we showed that KLF17 expression was decreased in lung adenocarcinoma. Reduced expression of KLF17 was correlated significantly with a short survival time in patients with lung adenocarcinoma (P<0.0001). Moreover, KLF17 expression was an independent prognostic indicator for patients with lung adenocarcinoma. KLF17 expression level was correlated with the tumor stage (P=0.016) and tumor size (P=0.001) in lung adenocarcinoma. Overexpression of KLF17 inhibited cell growth in A549 and PC-9 cell lines. In conclusion, it is possible that KLF17 inhibits tumor growth in lung adenocarcinoma. The reduced expression of KLF17 is a valuable prognostic indicator for patients with lung adenocarcinoma, and KLF17 could be a novel target for treatment of lung adenocarcinoma.

Targeting the CD134-CD134L interaction using anti-CD134 and/or rhCD134 fusion protein as a possible strategy to prevent lupus nephritis.

Lupus nephritis (LN) is characterized by an increased upregulation of Th1. This study was undertaken to evaluate the role of CD134 in cytokine production in peripheral blood mononuclear cells (PBMCs) from subjects with LN. Percentages of IFN-gamma- (Th1), IL-4-, and IL-10- (Th2) producing cells within the PBMC CD4+ T cell population of LN subjects were found to be higher than those of healthy subjects. Stimulation of PBMC from LN subjects with anti-CD3 epsilon mAb/rIL-2 resulted in further increases in cytokine production. Stimulation in the presence of anti-CD134 mAb resulted in reduced IL-4 and IL-10 production; however, it also resulted in increased IFN-gamma production. Stimulation in the presence of the fusion protein rhCD134:Fc resulted in decreased production of all three cytokines. The possibilities that anti-CD134 therapy may control the extent of IL-4- and IL-10-mediated damage in active LN and that rhCD134:Fc therapy may prevent occurrence of LN are discussed.

Krüppel-like factor 17 inhibits urokinase plasminogen activator gene expression to suppress cell invasion through the Src/p38/ MAPK signaling pathway in human lung adenocarcionma.

Krüppel-like factor 17 (KLF17) has been reported to be involved in invasion and metastasis suppression in lung cancer, but the molecular mechanisms underlying the anti-invasion and anti-metastasis roles of KLF17 in lung cancer are not fully illustrated. Here, we showed that KLF17 inhibited the invasion of A549 and H322 cells; the anti-invasion effect of KLF17 was associated with the suppression of urokinase plasminogen activator (uPA/PLAU) expression. KLF17 can bind with the promoter of uPA and inhibit its expression. Enforced expression of uPA abrogated the anti-invasion effect of KLF17 in A549 and H322 cells. In addition, immunohistochemistry staining showed that the protein expression of KLF17 was negatively correlated with that of uPA in archived samples from patients with lymph node metastasis of lung adenocarcinoma (rho = -0.62, P = 0.01). The mutually exclusive expression of KLF17 with uPA could predict lymph node metastasis for lung adenocarcinoma (AUC = 0.758, P = 0.005). Enforced expression of KLF17 inhibited the expression of phosphorylated Src and phosphorylated p38/MAPK in A549 and H322 cells. The invasiveness of the cells were suppressed by treating with sb203580 (p38/MAPK inhibitor) or HY-13805 (PP2, Src inhibitor). furthermore, p38/MAPK inhibition could block the KLF17-induced reduction of p-p38/MAPK and uPA, and Src inhibition enhanced the KLF17-induced suppression of p-Src and uPA in A549 and H322 cells. In conclusion, our study indicated that KLF17 suppressed the uPA-mediated invasion of lung adenocarcinoma. The Src and p38/MAPK signaling pathways were suggested as mediators of KLF17-induced uPA inhibition, thus providing evidence that KLF17 might be a potential anti-invasion candidate for lung adenocarcinoma.

[Analysis of virulence-related properties of P. gingivalis clinical isolates].

PURPOSE: To analyze the relation of virulence properties and pathogenicity of Porphyromonas gingivalis (P.gingivals) isolated from Chinese patients. METHODS: Based on the previous analysis of virulence properties, investigations of pathogenicity properties, including the adhesion to host cells, gingipain activities, and resistance to host immune reaction were performed to analyze the diversity of pathogenic properties of these strains. SAS 8.02 software package was used for statistical analyses. RESULTS: Less-virulent type strain showed higher adherence ability to host red blood cells and weaker resistance to serum killing, while virulent type strain showed the opposite reactions. However, the clinic isolates presented just the opposite relationship between virulence properties and pathogenicity. In addition, there was no obvious correlation between gingipain activities and virulence properties of P. gingivalis strains. CONCLUSIONS: The results suggested that there is no obvious linkage between the P. gingivalis virulence properties and pathogenicity diversity.

[Using DGGE to analyze bacteria community structure changes in subgingival plaque after periodontal initial therapy].

PURPOSE: To analyze bacteria community structure changes in subgingival plaque after initial therapy, and to provide experimental evidences for clinical decision. METHODS: Six patients with chronic periodontitis were chosen. Subgingival plaque samples, as well as clinical indexes, were collected at baseline and six weeks after initial therapy. The generated two different 16S rDNA fragments of subgingival plaque samples were separated by denaturing gel, creating bands patterns representative of community structure. Subsequent cluster analysis was made. RESULTS: The clinical indexes were improved significantly. The diversity of population of clinical subgingival samples was not significantly different between baseline and 6 weeks after initial therapy. Through cluster analysis, it was confirmed that same patient got similar subgingival plaque between baseline and 6 weeks after treatment. CONCLUSIONS: Same patient tend to get similar subgingival plaque between baseline and after initial therapy. DGGE can detect the microbial composition of subgingival plaque.

[Virulent properties of Porphyromonas gingivalis clinical strains].

PURPOSE: To investigate the virulent properties of P. gingivalis clinical strains. METHODS: By using mouse abscess model (MAM), six P. gingivalis clinical strains, L2, L3, L4, L5, L11 and L12 were subcutaneously inoculated into the central back of BALB/C mice. The clinical signs of the mice were observed and the virulent properties of the clinical strains were analyzed by comparison with type strains of W83 and ATCC 33277. SPSS11.5 software package was used for statistical analysis. RESULTS: According to the criteria established in previous reports, L3, ATCC33277, and W83 produced a localized abscess at the site of injection and were categorized as noninvasive. L2, L5, L11, and L12-induced lesions spread to distant organs and may produce severe systemic reactions and these strains were classified as invasive. CONCLUSIONS: Clinical strains from Chinese patients showed similarities to type strains in MAM. The virulent properties of P. gingivalis clinical strains are quite different from each other.

[The effects of recombinant porcine amelogenin on human gingival epithelial cells].

PURPOSE: To investigate the potential effect of recombinant 25kDa porcine amelogenin (rPAm) on attachment, proliferation and migration of primarily cultured human gingival epithelial cells (HGEC). METHODS: The second passage of HGECs were exposed to different concentrations of rPAm (0, 5, 10, 20µg/mL, respectively). Proliferation and attachment activities was measured by using cell counting method. Cellular migration was assayed by using an in vitro wound healing model. The data was quantified by the analysis of GraphPad Prism software. RESULTS: rPAm inhibited HGEC attachment in the adhesion assay, the effect was depended on time and rPAm dose. rPAm suppressed the growth rate of HGEC, that was also dose and time dependent. rPAm inhibited the migration ability of HGEC, the concentration of 20µg/mL group had the most significant effect. CONCLUSIONS: rPAm significantly inhibit the growth rate, cell adhesion and migration of HGEC, and the effect was dose- and time- dependent.

Effect of anti-CD134L mAb and CTLA4Ig on ConA-induced proliferation, Th cytokine secretion, and anti-dsDNA antibody production in spleen cells from lupus-prone BXSB mice.

We sought to evaluate the effects of combined downregulation of CD134 and cytotoxic T lymphocyte-associated antigen 4 (CTLA4) on the autoimmune process of lupus. Concanavalin A (ConA)-induced proliferation, T helper cell cytokine secretion, and anti-double stranded DNA (dsDNA) antibody production were measured in cultures of splenic lymphocytes derived from lupus-prone BXSB mice. Splenocytes from six prednisone-treated and six untreated male lupus-prone BXSB mice, as well as from six syngeneically normal C57BL/6 male mice, were stimulated with ConA. BXSB splenocytes from untreated mice were exposed to anti-CD134L mAb, CTLA4 linked to the Fc portion of IgG1 (CTLA4Ig), or both. The magnitude of splenocyte proliferation and the levels of IFN-gamma, IL-6, and anti-dsDNA antibody were: (1) significantly higher in cultures of ConA-stimulated control and other cells than in unstimulated cells, (2) similar in cultures of normal and BXSB cells treated with anti-CD134 and CTLA4Ig or prednisone and (3) significantly reduced in cultures of ConA-stimulated and unstimulated cells treated with anti-CD134L and CTLA4Ig or prednisone compared with cells treated with CD134L or CTLA4Ig alone. Like corticosteroids, anti-CD134L mAb or CTLA4Ig can inhibit T- and B-cell activation by blocking the CD134-CD134L or CD28/CTLA4-B7 co-stimulatory pathway. The combined immune intervention described herein may prove useful for the treatment of autoimmune diseases such as systemic lupus erythematosus.