An RGD containing peptide from HIV-1 Tat-(65-80) modulates protooncogene expression in human bronchoalveolar carcinoma cell line, A549.

被引:24
作者
ElSolh, A
Kumar, NM
Nair, MPN
Schwartz, SA
LwebugaMukasa, JS
机构
[1] SUNY BUFFALO,SCH MED & BIOMED SCI,BUFFALO GEN HOSP,DEPT INTERNAL MED,DIV PULM & CRIT CARE,BUFFALO,NY 14203
[2] SUNY BUFFALO,SCH MED & BIOMED SCI,BUFFALO GEN HOSP,DEPT INTERNAL MED,DIV ALLERGY & IMMUNOL,BUFFALO,NY 14203
[3] SUNY BUFFALO,SCH MED & BIOMED SCI,BUFFALO GEN HOSP,DEPT INTERNAL MED,LUNG BIOL RES PROGRAM,BUFFALO,NY 14203
关键词
D O I
10.3109/08820139709022692
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Tat (transactivator of transcription) is essential for HIV-1 replication in vivo and in vitro. Tat-(65-80), an RGD containing domain, has been shown to regulate proliferative function of a variety of cell lines, including a human adenocarcinoma cell line, A549. The exact cellular and molecular mechanisms by which these effects are mediated, remain unknown. To evaluate the hypothesis that Tat-(65-80) modulates the expression of immediate early genes (IEG) c-jun, c-myc, c-fos and the tumor suppressor gene p53, serum starved A549 cells were incubated with Tat-(65-80) or heat-inactivated Tat-(65-80) at 10 ng/ml. Total cellular RNA was Isolated from the cells at various time points (0-24 hours). In each case, 5 mu g of RNA was reverse transcribed in 20 mu 1 of reaction volume. Equal amounts of cDNA were subjected to polymerase chain reaction (PCR) and analyzed by electrophoresis. The photographic negatives of the ethidium bromide stained gels were quantitated by densitometric scanning and normalized to corresponding beta-actin PCR products. Treatment with Tat-(65-80) showed a twofold induction of c-jun at 0.5 h. Peak expression occurred at 60 minutes and remained above baseline at 24 hours (h). c-myc was increased at 0.5 h, reached a twofold increase at 2 h and remained above baseline at 24 h. c-Sos increased seven fold at 0.5 h and declined subsequently to baseline at 8 h, p-53 gene was reduced fivefold at 0.5 h and remained downregulated thereafter. These results show that Tat-(65-80) can modulate growth related genes in human lung epithelial cells.
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页码:351 / 370
页数:20
相关论文
共 38 条
[1]   THE TAT PROTEIN OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1, A GROWTH-FACTOR FOR AIDS KAPOSI-SARCOMA AND CYTOKINE-ACTIVATED VASCULAR CELLS, INDUCES ADHESION OF THE SAME CELL-TYPES BY USING INTEGRIN RECEPTORS RECOGNIZING THE RGD AMINO-ACID-SEQUENCE [J].
BARILLARI, G ;
GENDELMAN, R ;
GALLO, RC ;
ENSOLI, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (17) :7941-7945
[2]   IDENTIFICATION OF AN ARG-GLY-ASP (RGD) CELL-ADHESION SITE IN HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 TRANSACTIVATION PROTEIN, TAT [J].
BRAKE, DA ;
DEBOUCK, C ;
BIESECKER, G .
JOURNAL OF CELL BIOLOGY, 1990, 111 (03) :1275-1281
[3]   THE C-FOS PROTEIN INTERACTS WITH C-JUN/AP-1 TO STIMULATE TRANSCRIPTION OF AP-1 RESPONSIVE GENES [J].
CHIU, R ;
BOYLE, WJ ;
MEEK, J ;
SMEAL, T ;
HUNTER, T ;
KARIN, M .
CELL, 1988, 54 (04) :541-552
[4]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[5]   IDENTIFICATION AND NUCLEOTIDE-SEQUENCE OF A HUMAN LOCUS HOMOLOGOUS TO THE V-MYC ONCOGENE OF AVIAN MYELOCYTOMATOSIS VIRUS MC29 [J].
COLBY, WW ;
CHEN, EY ;
SMITH, DH ;
LEVINSON, AD .
NATURE, 1983, 301 (5902) :722-725
[6]   THE TRANSACTIVATOR GENE OF THE HUMAN T-CELL LYMPHOTROPIC VIRUS TYPE-III IS REQUIRED FOR REPLICATION [J].
DAYTON, AI ;
SODROSKI, JG ;
ROSEN, CA ;
GOH, WC ;
HASELTINE, WA .
CELL, 1986, 44 (06) :941-947
[7]  
DENIS M, 1994, J IMMUNOL, V153, P2072
[8]   TAT PROTEIN OF HIV-1 STIMULATES GROWTH OF CELLS DERIVED FROM KAPOSIS-SARCOMA LESIONS OF AIDS PATIENTS [J].
ENSOLI, B ;
BARILLARI, G ;
SALAHUDDIN, SZ ;
GALLO, RC ;
WONGSTAAL, F .
NATURE, 1990, 345 (6270) :84-86
[9]   RELEASE, UPTAKE, AND EFFECTS OF EXTRACELLULAR HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 TAT PROTEIN ON CELL-GROWTH AND VIRAL TRANSACTIVATION [J].
ENSOLI, B ;
BUONAGURO, L ;
BARILLARI, G ;
FIORELLI, V ;
GENDELMAN, R ;
MORGAN, RA ;
WINGFIELD, P ;
GALLO, RC .
JOURNAL OF VIROLOGY, 1993, 67 (01) :277-287
[10]   THE ROLE OF TAT IN THE HUMAN-IMMUNODEFICIENCY-VIRUS LIFE-CYCLE INDICATES A PRIMARY EFFECT ON TRANSCRIPTIONAL ELONGATION [J].
FEINBERG, MB ;
BALTIMORE, D ;
FRANKEL, AD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (09) :4045-4049