PURPOSE: It has become clear that, together with proliferation, deregulation of apoptosis plays a pivotal role in tumorigenesis, and the somatic mutations of apoptosis-related genes have been reported in human cancers. PUMA, a pro- apoptotic member of Bcl-2 family, mediates p53-deependent and -independent apoptosis. The aim of this study was to explore whether alteration of PUMA protein expression is a characteristic of human lung cancers. MATERIALS AND METHODS: To explore the possibility that the genetic alterations of PUMA might be involved in the development of human cancers, we analyzed the entire coding region and all splice sites of human PUMA gene in 100 human non-small cell lung cancers (NSCLCs) by polymerase chain reaction (PCR)-based single-strand conformation polymorphism (SSCP). RESULTS: The PCR-SSCP analysis detected no mutation in the entire coding regions and all splice sites of human PUMA gene in the 100 NSCLCs. CONCLUSION: The data presented here suggested that PUMA gene mutation may not contribute to the pathogenesis of human NSCLCs.
Apoptosis ; Carcinogenesis ; Carcinoma, Non-Small-Cell Lung ; Clinical Coding ; Humans ; Lung Neoplasms* ; Lung* ; Polymerase Chain Reaction ; Puma*

OBJECTIVES: Previous research has found that greater income inequality is related to problematic alcohol use across a variety of geographical areas in the USA and New York City (NYC). Those studies used self-reported data to assess alcohol use. This study examined the relationship between within-neighborhood income inequality and alcohol-related emergency department (ED) visits.METHODS: The study outcome was the alcohol-related ED visit rate per 10,000 persons between 2010 and 2014, using data obtained from the New York Statewide Planning and Research Cooperative System. The main predictor of interest was income inequality, measured using the Gini coefficient from the American Community Survey (2010-2014) at the public use microdata area (PUMA) level (n=55) in NYC. Variables associated with alcohol-related ED visits in bivariate analyses were considered for inclusion in a multivariable model.RESULTS: There were 420,568 alcohol-related ED visits associated with a valid NYC address between 2010 and 2014. The overall annualized NYC alcohol-related ED visit rate was 100.7 visits per 10,000 persons. The median alcohol ED visit rate for NYC PUMAs was 88.0 visits per 10,000 persons (interquartile range [IQR], 64.5 to 133.5), and the median Gini coefficient was 0.48 (IQR, 0.45 to 0.51). In the multivariable model, a higher neighborhood Gini coefficient, a lower median age, and a lower percentage of male residents were independently associated with the alcohol-related ED visit rate.CONCLUSIONS: This study found that higher neighborhood income inequality was associated with higher neighborhood alcohol-related ED visit rates. The precise mechanism of this relationship is not understood, and further investigation is warranted to determine temporality and to assess whether the results are generalizable to other locales.
Alcohol Drinking ; Emergencies ; Emergency Service, Hospital ; Humans ; Male ; Puma ; Residence Characteristics ; Socioeconomic Factors ; Surveys and Questionnaires ; Urban Health

BACKGROUND: Several lines of evidence have indicated that the deregulation of apoptosis is involved in the mechanisms of cancer development, and somatic mutations of the apoptosisrelated genes have been reported in human cancers. Members of the bcl-2 family proteins regulate the intrinsic apoptosis pathway mainly in the mitochondria. The aim of this study was to explore whether the somatic mutation of the proapoptotic bcl-2 family genes, one of the mechanisms that prolong the survival of cancer cells, occurred in colorectal carcinomas. METHODS: In the current study, to detect the somatic mutations in the DNA sequences encoding the bcl-2 homology 3 (BH3) domain of the human bak, bid, bik, bim, PUMA, bcl-rambo, bcl-G, and bmf genes in 98 colon adenocarcinomas, we used polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), and DNA sequencing. RESULTS: The SSCP analysis detected no evidence of somatic mutations of the genes in the coding regions of the BH3 domain in the cancers. CONCLUSIONS: The data presented here indicate that the proapoptotic bcl-2 family genes, bak, bid, bik, bim, PUMA, bcl-rambo, bcl-G and bmf may not be somatically mutated in human colorectal carcinomas, and suggest that the colorectal cancers may not utilize mutational events of these proapoptotic bcl-2 family genes in the mechanisms for evading apoptosis.
Adenocarcinoma ; Apoptosis ; Base Sequence ; Clinical Coding ; Colon* ; Colonic Neoplasms ; Colorectal Neoplasms ; Humans ; Mitochondria ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Puma ; Sequence Analysis, DNA

Plant-derived triterpenoids commonly possesses biological properties such as anti-inflammatory, anti-microbial, anti-viral and anti-cancer. Luvunga scandens is one of the plant that produced triterpenoids. The aims of the study was to analyze cell cycle profile and to determine the expression of p53 unregulated modulator of apoptosis (PUMA), caspase-8 and caspase-9 genes at mRNA level in MCF-7 cell line treated with two triterpenoids, flindissol (1) and 3-oxotirucalla-7,24-dien-21-oic-acid (2) isolated from L. scandens. The compounds were tested for cell cycle analysis using flow cytometer and mRNA expression level using quantitative RT-PCR. The number of MCF-7 cells population which distributed in Sub G1 phase after treated with compound 1 and 2 were 7.7 and 9.3% respectively. The evaluation of the expression of genes showed that both compounds exhibited high level of expression of PUMA, caspase-8 and caspase-9 as normalized to β-actin via activation of those genes. In summary, the isolated compounds of L. scandens plant showed promising anticancer properties in MCF-7 cell lines.
Apoptosis ; Caspase 8 ; Caspase 9 ; Cell Cycle Checkpoints* ; Cell Cycle* ; Flow Cytometry ; G1 Phase ; Gene Expression* ; MCF-7 Cells* ; Plants ; Puma ; RNA, Messenger

PURPOSE: Tacrolimus (FK506) has been widely used as an immunosuppressant in organ transplanted recipients to suppress organ rejection phenomenon. We investigated the role of oxidative stress and heme oxygense-1 by FK506 on human Jurkat T cells. METHODS: The cells viability was examined by DAPI stain, enzyme activity of caspase family proteins, and western blotting for Baks, PUMA, iNOS, HO-1. Cells were cultured in the absence or presence of CoPPIX or ZnPPIX and the fluorescence intensity was analyzed using a flow cytometry. RESULTS: Treatment with FK506 increased the generation of reactive oxygen species (ROS), including hydrogen peroxide and superoxide anion, and NO in Jurkat cells in a dose-dependent manner. Immunohistochemistry and Western blot analysis data revealed the hemoxygenase-1 (HO-1) was induced by the addition of FK506 in Jurkat cells. Induction of CoPP, HO-1 inducer, resulted in decreased intracellular H2O2 and NO concentrations. Instead ZnPP, an HO-1 competitive inhibitor did it reversely. In addition, ZnPP regulates iNOS protein synthesis by inhibition of HO-1. CONCLUSION: Increase of HO-1 expression would induce to decrease the intracellular H2O2 and NO concentrations. Also, HO-1 would regulate iNOS protein synthesis. Consequently, we can expect the regulation of HO-1 expression with concomitants use of FK506 to suppress organ rejection phenomenon by enhancing apoptosis.
Apoptosis ; Blotting, Western ; Flow Cytometry ; Fluorescence ; Heme ; Heme Oxygenase-1 ; Humans ; Hydrogen Peroxide ; Immunohistochemistry ; Indoles ; Jurkat Cells ; Lymphocytes ; Oxidative Stress ; Proteins ; Puma ; Reactive Oxygen Species ; Rejection (Psychology) ; Superoxides ; T-Lymphocytes ; Tacrolimus ; Transplants

PURPOSE: c-Met is an attractive potential target for novel therapeutic inhibition of human cancer, and c-Met tyrosine kinase inhibitors (TKIs) are effective growth inhibitors of various malignancies. However, their mechanisms in anticancer effects are not clear. In the present study, we investigated the possibility that blocking c-Met signaling induces p53-mediated growth inhibition in lung cancer. MATERIALS AND METHODS: The growth inhibitory effects of c-Met TKI (SU11274) on lung cancer cells and a xenograft model were assessed using the MTT assay, flow cytometry, and terminal deoxyribonucleotide transferase-mediated nick-end labeling staining. The role of p53 protein in the sensitivity of c-Met TKI (SU11274) was examined by Western blot analysis and immunohistochemistry. RESULTS: SU11274 significantly induced apoptosis in A549 cells with wild-type p53, compared with that in Calu-1 cells with null-type p53. SU11274 increased p53 protein by enhancing the stability of p53 protein. Increased p53 protein by SU11274 induced up-regulation of Bax and PUMA expression and down-regulation of Bcl-2 expression, subsequently activating caspase 3. In p53 knock-out and knock-in systems, we confirmed that SU11274 caused apoptosis through the p53-mediated apoptotic pathway. Likewise, in the A549 xenograft model, SU11274 effectively shrank tumor volume and induced apoptosis via increased p53 protein expression. Blocking c-Met signaling increased the level of p53 protein. CONCLUSION: Our finding suggested that p53 plays an important role in SU11274-induced apoptosis, and p53 status seems to be related to the sensitivity to SU11274 in lung cancer.
Apoptosis ; Blotting, Western ; Caspase 3 ; Down-Regulation ; Flow Cytometry ; Growth Inhibitors ; Humans ; Indoles ; Lung ; Lung Neoplasms ; Molecular Targeted Therapy ; Piperazines ; Protein-Tyrosine Kinases ; Puma ; Sulfonamides ; Transplantation, Heterologous ; Tumor Burden ; Tumor Suppressor Protein p53 ; Up-Regulation