Clinical and prognostic significance of Krüppel-like transcription factor 6 expression in 67 patients with cutaneous malignant melanoma in China.

Krüppel-like transcription factor 6 (KLF6) is a ubiquitous tumor suppressor gene involved in regulating cell growth, proliferation, differentiation and angiogenesis. The objective of the study was to investigate the clinical and prognostic significance of KLF6 expression in cutaneous malignant melanoma (CMM) patients. A total of CMM 67 patients were enrolled in this study. The specimens were evaluated by immunohistochemistry to detect KLF6. The positive KLF6 expression in CMM tissues was significantly lower than in normal skin tissues (p < 0.01).The presence of KLF6 in CMM was correlated with ulceration (p < 0.01), lymph node metastasis (p < 0.01) and clinical stage (p < 0.01). The overall 5-year survival rate of the 67 patients was 13.4%. The 5-year survival rate of patients with negative KLF6 expression was correlated with KLF6 expression (p < 0.01), ulceration (p < 0.01), lymph node metastasis (p < 0.01), clinical stage (p < 0.01) and operation type (p < 0.01). Ulceration and clinical staging were independent relevant factors. In conclusion, the presence of KLF6 in CMM was correlated with ulceration, lymph node metastasis, clinical stage and poor prognosis.

NPS - 2143 (hydrochloride) inhibits melanoma cancer cell proliferation and induces autophagy and apoptosis.

Melanoma is a common and aggressive skin cancer caused by the oncogenic transformation of melanocytes. NPS-2143 (hydrochloride) is a calcification drug that acts as an antagonist of the calcium-sensing receptor (CaSR) and consequently stimulates the release of parathyroid hormone. In the present work, we treated cells from the human melanoma cell line M14 to investigate the effects of NPS-2143 on melanoma cells and elucidate their underlying mechanisms. We observed that NPS-2143 inhibits the survival and proliferation of M14 cells and suppresses the migration and proliferation of M14 cells by inducing apoptosis. The Bax/Bcl­2 ratio in M14 cells was enhanced by the NPS-2143 treatment, suggesting that the mitochondrial apoptotic pathway was activated. The expression and phosphorylation of proteins involved in the PI3K signaling pathway were altered by NPS-2143 treatment. Our data show that NPS-2143 impacts the viability and induces the apoptosis of melanoma M14 cells through its impact on the PI3K signaling pathway. It suggests that NPS-2143 could represent a promising candidate for melanoma treatment.

Knock-down of filaggrin influences the mitogen-activated protein kinases signaling pathway in normal human epidermal keratinocytes.

BACKGROUND: Filaggrin is an essential structural protein of the stratum corneum binding to the keratin intermediate filaments to form a dense protein-lipid matrix. However, the function of filaggrin in epidermal terminal differentiation is not completely understood. AIM: To evaluate the effects of filaggrin on normal human epidermal keratinocytes (NHEKs) and to investigate the relevant mechanisms. METHODS: Short hairpin RNA (shRNA) technology was used to knock-down filaggrin in normal human epidermal keratinocytes (NHEKs). Western blot and real-time quantitative PCR (qRT-PCR) were performed to detect expression of filaggrin, differentiation-related proteins and MAPK-related proteins. RESULTS: Filaggrin was successfully knocked down in NHEKs (99% efficiency). We found that the lack of filaggrin significantly decreased the expression of some differentiation-related proteins, including Cytokeratin 5 protein, Cytokeratin 14 protein, ST14 protein and SPRR3 protein (P<0.05). In addition, filaggrin knock-down significantly decreased expression of p-p38, p-ERK1/2, p-JNK, p-Akt, and p-NF-κB in NHEKs. CONCLUSION: Our study shows that filaggrin regulates epidermal terminal differentiation and impairs MAPK signaling pathway in normal human epidermal keratinocytes.

MAPK Pathway Involved in Epidermal Terminal Differentiation of Normal Human Epidermal Keratinocytes.

OBJECTIVE: To investigate the effect of mitogen-activated protein kinase (MAPK) signaling pathway in epidermal terminal differentiation. METHODS: The MAPK pathways (p38, ERK1/2, JNK) were inhibited by SB203580, PD98059, and SP600125 in normal human epidermal keratinocytes (NHEKs), respectively. Western blotting assays were performed to detect expression of filaggrin and differentiation-related proteins. The mRNA expressions of differentiation-related proteins were detected by real-time quantitative PCR (qRT-PCR). RESULTS: Inhibition of MAPK pathway by SB203580, PD98059, and SP600125 resulted in significant reduction of filaggrin expression in NHEKs. Inhibition of the p38 MAPK pathway decreased the expression of differentiation-related proteins (cytokeratin 5, cytokeratin 14, ST14, and SPRR3), Akt, and NF-κB. Inhibition of JNK also suppressed expression of cytokeratin 14, SPRR3, Akt, and NF-κB. However, inhibition of ERK1/2 merely decreased expression of SPRR3 and Akt. CONCLUSION: MAPK pathways regulates epidermal terminal differentiation in NHEKs. The p38 signaling pathway plays an especially important role.

The impact of dual-source parallel radiofrequency transmission with patient-adaptive shimming on the cardiac magnetic resonance in children at 3.0 T.

The cardiac magnetic resonance (CMR) of children at 3.0 T presents a unique set of technical challenges because of their small cardiac anatomical structures, fast heart rates, and the limited ability to keep motionless and hold breathe, which could cause problems associated with field inhomogeneity and degrade the image quality. The aim of our study was to evaluate the effect of dual-source parallel radiofrequency (RF) transmission on the B1 homogeneity and image quality in children with CMR at 3.0 T. The study was approved by the institutional ethics committee and written informed consent was obtained. A total of 30 free-breathing children and 30 breath-hold children performed CMR examinations with dual-source and single-source RF transmission. The B1 homogeneity, contrast ratio (CR) of cine images, and off-resonance artifacts in cine images between dual-source and single-source RF transmission were assessed in free-breathing and breath-hold groups, respectively. In both free-breathing and breath-hold groups, higher mean percentage of flip angle (free-breathing group: 104.2 ±â€Š4.6 vs 95.5 ±â€Š6.3, P < .001; breath-hold group: 101.5 ±â€Š5.1 vs 92.5 ±â€Š6.3, P < .001) and lower coefficient of variation (free-breathing group: 0.06 ±â€Š0.02 vs 0.09 ±â€Š0.03, P < .001; breath-hold group: 0.07 ±â€Š0.03 vs 0.10 ±â€Š0.04, P = .005) were found with dual-source than with single-source RF transmission. Both the CRs in the horizontal long axis (HLA) and short axis of cine images with dual-source RF transmission was improved (P < .05 for all). The scores of off-resonance artifacts in the HLA with dual-source RF transmission were higher in both free-breathing and breath-hold groups (P < .05 for all), with substantial interreader agreement (kappa values from 0.68 to 0.74). Compared with conventional single-source, dual-source parallel RF transmission could significantly improve the B1 homogeneity and image quality for CMR in children at 3.0 T. This technology could be taken into account in CMR for children with cardiac diseases.

Design, Synthesis, and Anti-HIV Evaluation of Novel Triazine Derivatives Targeting the Entrance Channel of the NNRTI Binding Pocket.

A novel series of triazine derivatives targeting the entrance channel of the HIV-1 non-nucleoside reverse transcriptase inhibitor binding pocket (NNIBP) were designed and synthesized on the basis of our previous work. The results of a cell-based antiviral screening assay indicated that most compounds showed good-to-moderate activity against wild-type HIV-1 with EC50 values within the concentration range of 0.0078-0.16 µm (compound DCS-a4, EC50  = 7.8 nm). Some compounds displayed submicromolar activity against the K103N/Y181C resistant mutant strain (such as compound DCS-a4, EC50  = 0.65 µm). Molecular modeling studies confirmed that the new compounds could bind into the NNIBP similarly as the lead compound, and the newly introduced flexible heterocycles could occupy the entrance channel effectively. In addition, the preliminary structure-activity relationship and the RT inhibitory assay are presented in this study.

Expression of hepatocyte growth factor and c-Met in non-small-cell lung cancer and association with lymphangiogenesis.

Previous experimental studies have demonstrated that hepatocyte growth factor (HGF) and its receptor c­Met serve an important function in lymphangiogenesis, but their biological functions in malignant tumors have remained elusive. The present study aimed to investigate the expression patterns of HGF­α and c­Met and their association with vascular endothelial growth factor (VEGF)­C, lymphatic vessel density and lymph node metastasis in non­small­cell lung cancer (NSCLC). In the present study, the lymphatic microvessel density (LMVD) and the expression levels of HGF­α, its receptor c­Met and VEGF­C were determined in 113 human NSCLC tissues and 113 normal lung tissue samples, using immunohistochemical staining. As a result, it was determined that the expression levels of HGF­α, c­Met and VEGF­C were significantly higher in NSCLC tissues than those in normal lung tissues (HGF­α, 67.3 vs. 20.4%, P<0.001; c­Met, 74.3 vs. 23.0%, P<0.001; and VEGF­C, 65.5 vs. 23.9%, P<0.001). HGF­α expression was observed to be significantly associated with that of VEGF­C (r=0.234, P=0.012) or c­Met (r=0.648, P<0.001). In addition, there was a positive correlation between the expression levels of VEGF­C and c­Met (r=0.224, P=0.017). In NSCLC tissues, the expression of HGF­α, c­Met or VEGF­C was significantly correlated with the LMVD (P=0.045, 0.002 and 0.001, respectively), and lymph node metastasis was more common in HGF­α, c­Met or VEGF­C­positive groups (P=0.020, 0.020 and 0.009, respectively). In addition, the HGF­α or VEGF­C­positive groups presented shorter survival time periods. In conclusion, the expression of HGF­α or c­Met was closely correlated with VEGF­C, LMVD and metastases of lymph nodes, indicating that HGF­α, c­Met and VEGF­C may perform important and collaborative actions in lymphangiogenesis and lymphatic metastasis of primary NSCLC.

Overexpression of both platelet-derived growth factor-BB and vascular endothelial growth factor-C and its association with lymphangiogenesis in primary human non-small cell lung cancer.

BACKGROUND: Metastatic spread of tumor through lymphatic vasculature is an important adverse prognostic factor in a variety of human cancer and tumor lymphangiogenesis requires the interplay of several growth factors. Platelet-derived growth factor (PDGF)-BB and vascular endothelial growth factor (VEGF)-C are two important molecules involving in tumor metastasis and lymphangiogenesis. Therefore, the aim of this study was to investigate the coexpression of PDGF-BB and VEGF-C in primary human non-small cell lung cancer (NSCLC) and its association with lymphangiogenesis. METHODS: Using immunohistochemical staining, PDGF-BB and VEGF-C expression were detected in 109 primary NSCLC tissues, while the lymphatic micro-vessel density (LMVD) was counted. RESULTS: Of 109 cases, PDGF-BB and VEGF-C overexpression was 66.97% (73/109) and 65.14% (71/109), respectively. 52 (47.7%) had overexpression of both PDGF-BB and VEGF-C (P+V+), 21 (19.3%) overexpression of PDGF-BB but low expression of VEGF-C (P+V-), 19(17.4%) overexpression of VEGF-C but low expression of PDGF-BB (P-V+) and 17(15.6%) low expression of both PDGF-BB and VEGF-C (P-V-). PDGF-BB expression was positively related to that of VEGF-C (r=0.451, p=0.034). LMVD in cases with P+V+was much higher than those with P-V- (p=0.004). In addition, the patients with P+V+were younger and also had larger tumor size, more likely lymph node metastasis and worse histological differentiation than those with P-V-. Moreover, the overall survival (OS) of patients with P+V+was shorter than those with P-V- (p=0.015). CONCLUSION: Coexpression of both PDGF-BB and VEGF-C was associated with lymphangiogenesis and poor prognosis in NSCLC, and might play a critical role in NSCLC progression. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/2261801312571320.

Thrombin promotes airway remodeling via protease-activated receptor-1 and transforming growth factor-ß1 in ovalbumin-allergic rats.

BACKGROUND: Protease-activated receptor-1 (PAR-1) is widely distributed in platelets and involved in coagulation cascade activated by thrombin. In this study, we intend to explore the role of PAR-1 in the process of thrombin-inducing transforming growth factor-ß1 (TGF-ß1) to promote airway remodeling in ovalbumin (OVA)-allergic rats. MATERIALS AND METHODS: A rat model of chronic asthma was set up by systemic sensitization and repeated challenge to OVA. The doses of thrombin, recombinant hirudin, PAR-1 inhibitor ER-112780-06 varied for different groups. We evaluated the bronchoalveolar lavage fluid (BALF) concentration of thrombin in these groups. The protein and gene expression of PAR-1 was assessed and the expression of TGF-ß1 was also detected. RESULTS: The PAR-1 mRNA level and the protein level were higher in the airway of asthmatic rats than those of normal rats, and were significantly increased by thrombin treatment but decreased by thrombin-inhibitor treatment. Airway remodeling was strengthened by thrombin but weakened by thrombin inhibitor and PAR-1 antagonist. Expression of TGF-ß1 protein in asthmatic rats was significantly increased by thrombin treatment and decreased by thrombin-inhibitor treatment and PAR-1 antagonist treatment. CONCLUSION: The expression of PAR-1 is regulated by thrombin that induces the expression of TGF-ß1 to promote airway remodeling via PAR-1 in OVA-allergic rats.

Synthesis and biological activities of a 3'-azido analogue of Doxorubicin against drug-resistant cancer cells.

Doxorubicin (DOX), an anthracycline antibiotic, is one of the most active anticancer chemotherapeutic agents. The clinical use of DOX, however, is limited by the dose-dependant P-glycoprotein (P-gp)-mediated resistance. Herein, a 3'-azido analogue of DOX (ADOX) was prepared from daunorubicin (DNR). ADOX exhibited potent antitumor activities in drug-sensitive (MCF-7 and K562) and drug-resistant cell lines (MCF-7/DNR, K562/DOX), respectively. The drug resistance index (DRI) values of ADOX were much lower than that of DOX. The cytotoxicity experiments of ADOX or DOX against K562/DOX, with or without P-gp inhibitor, indicated that ADOX circumvents resistance by abolishing the P-gp recognition. This conclusion was further supported by drug influx/efflux flow cytometry experiments, as well as by molecular docking of ADOX to P-gp. In vivo animal tests, ADOX exhibited higher activity and less toxicity than DOX. The current data warranted ADOX for additional pre-clinical evaluations for new drug development.