Circulating lung cancer biomarkers: From translational research to clinical practice.

Fundamental studies on biomarkers as well as developed assays for their detection can provide valuable information facilitating clinical decisions. For patients with lung cancer, there are established circulating biomarkers such as serum progastrin-releasing peptide (ProGRP), neuron-specific enolase (NSE), squamous cell carcinoma antigen (SCC-Ag), carcinoembryonic antigen (CEA), and cytokeratin-19 fragment (CYFRA21-1). There are also molecular biomarkers for targeted therapy such as epidermal growth factor receptor (EGFR) gene, anaplastic lymphoma kinase (ALK) gene, KRAS gene, and BRAF gene. However, there is still an unmet need for biomarkers that can be used for early detection and predict treatment response and survival. In this review, we describe the lung cancer biomarkers that are currently being used in clinical practice. We also discuss emerging preclinical and clinical studies on new biomarkers such as omics-based biomarkers for their potential clinical use to detect, predict, or monitor subtypes of lung cancer. Additionally, between-method differences in tumor markers warrant further development and improvement of the standardization and harmonization for each assay.

The prognostic role of platelet-to-lymphocyte ratio in patients with acute heart failure: A cohort study.

Identification of rapid, inexpensive, and reliable prognostic factors can improve survival estimation and guide healthcare in patients with acute heart failure (AHF). In this study, we aimed to determine the prognostic value of the platelet-to-lymphocyte ratio (PLR) in patients with AHF. A total of 443 patients from two hospitals met the inclusion criteria from January 2010 to December 2017. Univariate and multivariate Cox analyses were performed to determine the association of PLR with survival. All-cause mortality was analysed using the Kaplan-Meier method. The 6-month survival rate for patients according to PLR quartiles (<110.63, 110.63-139.23, 139.23-177.17, and >177.17) were 90.09%, 76.79%, 50.07%, and 37.27%, respectively (p < 0.001). Univariate analysis identified high PLR (>110.63), old age (≥73 years), smoking habit, low estimated glomerular filtration rate (<57), and high platelet count (≥198 × 109/l) as poor prognostic factors for survival. In the multivariate analysis, after adjusting for confounding factors, the third (hazard ratio [HR] = 3.118, 95% confidence interval [CI] = 1.668-5.386, p < 0.001) and fourth (HR = 2.437, 95% CI = 1.302-3.653, p < 0.001) quartiles of PLR were identified as independent prognostic factors in patients with AHF. A higher PLR was associated with poor clinical outcomes in patients with AHF and might be a novel marker in AHF management.

Notch1 promotes the pericyte-myofibroblast transition in idiopathic pulmonary fibrosis through the PDGFR/ROCK1 signal pathway.

The goals of this study were to investigate the role of the Notch1/PDGFRß/ROCK1 signaling pathway in the pathogenesis of pulmonary fibrosis and to explore the possibility of treating fibrosis by targeting Notch1. Lung tissues from patients with pulmonary fibrosis were examined for the expression of Notch1/PDGFRß/ROCK1 using RT-qPCR, western blotting, and immunostaining. Cultured mouse lung pericytes were transfected with Notch1-overexpressed vectors or shRNA targeting PDGFRß/ROCK1 to examine cell behaviors, including proliferation, cell cycle arrest, and differentiation toward myofibroblasts. Finally, a mouse pulmonary fibrosis model was prepared, and a Notch1 inhibitor was administered to observe tissue morphology and pericyte cell behaviors. Human pulmonary fibrotic tissues presented with overexpression of Notch1, PDGFRß, and ROCK1, in addition to a prominent transition of pericytes into myofibroblasts. In cultured mouse lung pericytes, overexpression of Notch1 led to the accelerated proliferation and differentiation of cells, and it also increased the expression of the PDGFRß and ROCK1 proteins. The knockdown of PDGFRß/ROCK1 in pericytes remarkably suppressed pericyte proliferation and differentiation. As further substantiation, the administration of a Notch1 inhibitor in a mouse model of lung fibrosis inhibited the PDGFRß/ROCK1 pathway, suppressed pericyte proliferation and differentiation, and alleviated the severity of fibrosis. Our results showed that the Notch1 signaling pathway was aberrantly activated in pulmonary fibrosis, and this pathway may facilitate disease progression via mediating pericyte proliferation and differentiation. The inhibition of the Notch1 pathway may provide one promising treatment strategy for pulmonary fibrosis.

Integrated analysis of DNA methylation profiling and gene expression profiling identifies novel markers in lung cancer in Xuanwei, China.

BACKGROUND: Aberrant DNA methylation occurs frequently in cancer. The aim of this study was to identify novel methylation markers in lung cancer in Xuanwei, China, through integrated genome-wide DNA methylation and gene expression studies. METHODS: Differentially methylated regions (DMRs) and differentially expressed genes (DEGs) were detected on 10 paired lung cancer tissues and noncancerous lung tissues by methylated DNA immunoprecipitation combined with microarray (MeDIP-chip) and gene expression microarray analyses, respectively. Integrated analysis of DMRs and DEGs was performed to screen out candidate methylation-related genes. Both methylation and expression changes of the candidate genes were further validated and analyzed. RESULTS: Compared with normal lung tissues, lung cancer tissues expressed a total of 6,899 DMRs, including 5,788 hypermethylated regions and 1,111 hypomethylated regions. Integrated analysis of DMRs and DEGs identified 45 tumor-specific candidate genes: 38 genes whose DMRs were hypermethylated and expression was downregulated, and 7 genes whose DMRs were hypomethylated and expression was upregulated. The methylation and expression validation results identified 4 candidate genes (STXBP6, BCL6B, FZD10, and HSPB6) that were significantly hypermethylated and downregulated in most of the tumor tissues compared with the noncancerous lung tissues. CONCLUSIONS: This integrated analysis of genome-wide DNA methylation and gene expression in lung cancer in Xuanwei revealed several genes regulated by promoter methylation that have not been described in lung cancer before. These results provide new insight into the carcinogenesis of lung cancer in Xuanwei and represent promising new diagnostic and therapeutic targets.

[Relativity of nuclear factor-kappaB (P65/Rel-A) and angiotensin II type 1 receptor expression in early stage of lesions of adriamycin nephrosis in young rats and the effects of intervention].

OBJECTIVE: To investigate the trend and potential pathogenic role of nuclear factor (NF)-kappaB P(65)/Rel-A mRNA and angiotensin-II (AngII) receptor type 1 (AT(1)) proteins expression, and the relativity between them in early stage of renal tubulointerstitial lesions in young rats with adriamycin nephrosis and the interfering effects of treatment with angiotensin converting enzyme inhibitor (ACEI) benazepril and ACEI combined with AngII type 1 receptor antagonist (AT(1)RA) Losartan. METHODS: Male young Wistar rats with adriamycin nephrosis were used as experimental models. At different time points (weeks 1, 2, and 3 in early nephritic phase, the urinary protein and blood biochemical parameters were measured, and P(65)/Rel-A mRNA was detected; AT(1) protein expression was determined by in situ hybridization and immunohistochemical methods. The relativity between them was evaluated. RESULTS: In the early phase of tubulointerstitial lesions, following adriamycin injection and proteinuria aggravated progressively, at week 3, the proteinuria level had reached heavy proteinuria (123.2 +/- 7.7 mg/24 h). The serum parameters reflecting renal function were elevated. The inflammatory cells infiltrated into renal tissues, especially in tubulointerstitial regions, were increased markedly. Swelling of tubular epithelial cells, broadened tubulointerstitial areas, and protein casts in tubule were observed. In situ hybridization and immunochemical staining showed that AT(1) protein was expressed in tubular epithelial cell cytoplasm and on nuclear membranes (AT(1): 1st week 19.8 +/- 1.1%, 2nd week 25.0 +/- 2.6%, 3rd week 37.1 +/- 1.0% (control: 10.3 +/- 0.8%, 10.4 +/- 1.6%, 10.2 +/- 1.5%); and P(65)/Rel-A mRNA expression in the same locations was upregulated. P(65)/Rel-A translocation from cytoplasm into nucleus increased markedly simultaneously. The positive signal of hybridization dominated in cytoplasm gradually became dominant in the nuclei as the pathological changes progressed. The semiquantitative expression of P(65)/Rel-A was 24.0 +/- 3.3% at week 1, 34.2 +/- 2.4% at week 2, 39.9 +/- 6.4% at week 3, while the values of controls were 8.5 +/- 0.4%, 8.7 +/- 1.0%, and 8.4 +/- 0.9%, respectively. There was a positive correlation between AT(1) and P(65)/Rel-A expression in localization and time phase (r = 0.857, P < 0.01). However, the tendency of those factor's expression was all decreased in each treated group, the semiquantitative results were AT(1): 14.6 +/- 2.1%, 13.7 +/- 2.3%, 11.4 +/- 1.1%; P(65)/Rel-A: 18.5 +/- 3.4%, 22.8 +/- 1.6%, 26.7 +/- 4.9% at 1, 2, 3 weeks in ACEI treated group; AT(1): 12.4 +/- 1.5%, 11.1 +/- 1.0%, 10.3 +/- 0.8%; P(65)/Rel-A: 17.9 +/- 5.0%, 21.3 +/- 6.0%, 22.5 +/- 2.5% in AT(1)RA (Losartan) group, respectively. The significant difference were observed between all groups in different time points (P < 0.05). CONCLUSIONS: The present study suggested that NF-kappaB (P(65)/Rel-A) mRNA expression and its activity was enhanced significantly that synchronized with aggravating injures in tubulointerstitial lesions initial period induced by proteinuria-loading in nephrotic young rats. This tendency was related with AngII and its receptors system that may accelerate lesions progressing in many renal diseases.

Activation of intestinal arginine transport by protein kinase C is mediated by mitogen-activated protein kinases.

L-Arginine uptake by the small intestine can play a pivotal role in regulating nitric oxide synthesis and immune functions in catabolic states. We previously showed that protein kinase C (PKC) activation stimulates intestinal brush-border membrane arginine transport. However, the signaling pathways implicated in this activation have not been studied. The purpose of this study was to investigate the intracellular signal transduction pathways involved in the protein kinase C stimulation of arginine transport across the apical membrane of intestinal epithelial Caco-2 cells. [3H]-L-arginine transport activity, Northern blot analysis of mRNA levels of the intestinal arginine transporter CAT1, and Western blot analysis of the mitogen-activated protein (MAP) kinases phospho-p44/42 activity and phospho-MEK1/2 were measured in cultured Caco-2 cells treated with phorbol ester (phorbol 12-myristate 13-acetate, TPA; 0 to 0.5 micromol/L), and the MEK1 inhibitor PD 98059 (0 to 50 micromol/L). Phorbol ester stimulated intestinal arginine transport activity. Arginine transporter gene CAT1 mRNA, phospho-p44/42, and phospho-MEK1/2 levels were stimulated in phorbol ester-treated cells, compared with the control group. Phorbol ester stimulation of arginine transport activity and transporter CAT1 mRNA levels was blocked by PD 98059. These data suggest that phorbol ester stimulates arginine transport in Caco-2 cells via signaling pathways that lead to increased transcription and/or stabilization of CAT1 mRNA. Protein kinase C and MAP kinases MEK1/2 and p44/42 are key intracellular regulators involved in this signal transduction cascade.