1: Br J Dermatol. 2004 Sep;151(3):546-56. 

BCL2 and JUNB abnormalities in primary cutaneous lymphomas.

Mao X, Orchard G, Lillington DM, Child FJ, Vonderheid EC, Nowell PC, Bagot M,
Bensussan A, Russell-Jones R, Young BD, Whittaker SJ.

Skin Tumour Unit, St John's Institute of Dermatology, St Thomas' Hospital,
London SE1 7EH, UK. mxmayo@yahoo.co.uk

BACKGROUND: BCL2 is upregulated in nodal and extranodal B-cell non-Hodgkin's
lymphomas, with a consequent antiapoptotic effect. However, loss of BCL2 has
also been noted in some malignancies, suggesting a different molecular
pathogenesis. OBJECTIVES: To investigate genomic and protein expression status
of BCL2 and to compare the results with that of JUNB in primary cutaneous
lymphomas (PCLs). METHODS: We analysed gene copy number of BCL2 and JUNB in 88
DNA samples from 80 patients with PCL consisting of Sezary syndrome/mycosis
fungoides (SS/MF), primary cutaneous B-cell lymphoma (PCBCL) and primary
cutaneous CD30+ anaplastic large cell lymphoma (C-ALCL) by the use of real-time
polymerase chain reaction (PCR) and immunohistochemistry (IHC). Real-time PCR
and IHC findings were subsequently compared with the results of additional
fluorescent in situ hybridization (FISH) analysis of 23 cases of SS and
Affymetrix cDNA expression microarray study of two primary cutaneous T-cell
lymphoma (CTCL) cell lines. RESULTS: Real-time PCR analysis showed loss of BCL2
gene copy number in 22 of 80 PCL cases (28%), including 17 of 42 SS/MF, three of
13 C-ALCL and two of 33 PCBCL samples, and gain of BCL2 in four PCBCL samples.
Gain of JUNB was identified in 18 of 71 PCL cases (25%), including nine of 35
SS/MF, seven of 13 C-ALCL and two of 31 PCBCL samples. IHC analysis revealed
absent nuclear expression of BCL2 protein in 47 of 73 PCL cases, comprising 28
of 36 SS/MF, eight of eight C-ALCL and 11 of 29 PCBCL cases. In contrast, BCL2
protein expression was detected in 26 of 73 PCL cases, consisting of 18 of 29
PCBCL and eight of 36 SS/MF cases. JUNB protein expression was present in tumour
cells from 30 of 33 of SS/MF and eight of eight C-ALCL, and was absent in tumour
cells from 18 of 27 PCBCL cases. A comparison between BCL2 and JUNB revealed
loss of BCL2 and gain of JUNB in five of 35 SS/MF samples, and expression of
JUNB protein and absent BCL2 expression in 25 SS/MF and eight of eight C-ALCL
cases. In contrast, expression of BCL2 and absent JUNB expression were detected
in 67% of PCBCL cases. Additional FISH analysis revealed deletion of BCL2 in 19
of 23 SS cases (83%), including eight cases with BCL2 loss shown by real-time
PCR. Furthermore, Affymetrix expression microarray demonstrated decreased
expression of proapoptotic and antiapoptotic genes involved in BCL2 signalling
pathways such as BOK, BIM, HRK, RASA1 and STAT2 in two CTCL cell lines with BCL2
loss and absent BCL2 expression. Increased expression of JUNB was also
identified in the MF cell line. CONCLUSIONS: These findings provide a
comprehensive assessment of BCL2 and JUNB status in PCL, and suggest that there
is a selection pressure in a subset of CTCL cases for tumour cells showing BCL2
loss and upregulation of JUNB primarily through chromosomal deletion and
amplification, respectively.

PMID: 15377339 [PubMed - indexed for MEDLINE]


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2: J Neurochem. 2004 Jul;90(2):297-308. 

Regulation of RANTES/CCL5 expression in human astrocytes by interleukin-1 and
interferon-beta.

Kim MO, Suh HS, Brosnan CF, Lee SC.

Department of Pathology, Albert Einstein College of Medicine, Bronx, New York
10461, USA.

In the CNS, astrocytes are significant sources of RANTES/CCL5 (regulated upon
activation, normal T cell expressed and secreted), a CC-chemokine with important
biological function. Astrocyte RANTES/CCL5 has been shown to be induced by
interleukin-1 (IL-1), with interferon-gamma (IFNgamma) as a primer, but whether
type I interferons play any role in the expression of RANTES/CCL5 is not known.
In this report, we studied the detailed mechanism of RANTES/CCL5 induction in
primary human astrocytes activated with IL-1 and IFNbeta. Ribonuclease
protection assay and ELISA showed that IFNbeta, although not effective alone,
increased IL-1-induced RANTES/CCL5 expression, but did not antagonize IFNgamma.
IL-1 or IL-1/IFNbeta-induced RANTES/CCL5 expression was inhibited by the
super-repressor IkappaBalpha or inhibitors of p38 or c-Jun N-terminal kinase
(JNK) MAPKs (mitogen-activated protein kinases), but not by extracellular signal
regulated kinases (ERK) inhibitors. IFNbeta enhanced IL-1-induced
phosphorylation of p38 MAPK, but was not effective alone. Transfection with
mutated RANTES/CCL5 promoter-reporter constructs revealed that kappaB,
interferon-stimulated response element (ISRE) and CAATT-enhancer binding
protein-beta (C/EBPbeta) sites all contributed to IL-1/IFNbeta-induced
RANTES/CCL5 transcription. IFNbeta synergized with IL-1 to induce nuclear
accumulation of C/EBPbeta protein. They also synergized to form nuclear ISRE
complexes with Stat1, Stat2 and interferon regulatory factor-1 (IRF-1) proteins.
Together, our results demonstrate that IFNbeta plays a positive regulatory role
in the expression of RANTES/CCL5 in human astrocytes through several distinct
mechanisms.

PMID: 15228586 [PubMed - indexed for MEDLINE]


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3: Leuk Lymphoma. 2002 Jun;43(6):1289-95. 

Gene expression profiling in chronic myeloid leukemia patients treated with
hydroxyurea.

Bruchova H, Borovanova T, Klamova H, Brdicka R.

Institute of Hematology and Blood Transfusion, Department of Molecular Genetics,
Prague, Czech Republic.

Using array technology that allows the simultaneous detection of gene expression
of hundreds of genes, four patients with chronic myeloid leukemia (CML) were
investigated at diagnosis and after starting administration of hydroxyurea. To
detect the gene expression of peripheral blood mononuclears and granulocytes
Human Cancer cDNA Array (CLONTECH) with 588 gene probes was used. Gene
expression mononuclear and granulocyte profiles of patients at diagnosis were
compared with the control profiles. The significant expression changes observed
in most patients seemed to be important. Increased expression of c-jun
N-terminal kinase 2 (JNK2), integrin alpha E, MMP-8, MMP-9 was detected in both
fractions of most patients. In some samples PCNA, HDGF, MAPK p38, CD59 increased
expressions were found. Significant down-regulation of expression in patients
was detected in genes CDK4 inhibitor A, PURA, notch1 in mononuclears; STAT2,
STAT5, RAR-alpha, MCL-1, junB, caspase 4 in granulocytes; CDK6, GADD153, ERBB-3,
cadherin 5 in both fractions. Expression profiles detected in patients at
diagnosis did not differ markedly from those after one-week treatment with
hydroxyurea. Only in a few genes were significant changes after hydroxyurea
administration observed and inter-individual expression differences were rather
common.

PMID: 12152998 [PubMed - indexed for MEDLINE]


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4: J Immunol. 1994 Nov 15;153(10):4573-82. 

E1A repression of IL-6-induced gene activation by blocking the assembly of IL-6
response element binding complexes.

Takeda T, Nakajima K, Kojima H, Hirano T.

Division of Molecular Oncology, Osaka University Medical School, Japan.

Some viral products interfere with host antiviral defense mechanisms. Adenovirus
E1A represses IFN signal transduction pathways which induces gene activation and
an antiviral state. Both IFN and IL-6 activate Jak/Tyk protein tyrosine kinases
and the STAT (signal transducer and activator of transcription) family proteins.
We showed that 12S E1A repressed IL-6 signals activating the junB promoter and
the two IL-6 response elements (REs), JRE-IL6 and type II IL-6 RE (also called
acute phase response element), required for IL-6-induced activation of the junB
promoter and the type II acute phase reactant genes, respectively, in
hepatocytes. Conserved region 1 of the 12S E1A was responsible for the
repression. Target molecules of the repression by E1A appeared to be
IL-6-inducible DNA-binding proteins acting on the IL-6 REs. In a rat 3Y1 cell
line stably expressing E1A, the levels of IL-6-induced IL-6 RE binding proteins
were severely reduced compared with those in a parental 3Y1 cell line. Moreover,
we found that the levels of the STAT family proteins including Stat1-alpha
(p91), Stat1-beta (p84), Stat2 (p113), and Stat3 were decreased by the stable
expression of adenovirus E1A. The E1A-induced reduction in the amount of
DNA-binding proteins seemed to be partly responsible for the decreased
transcriptional activity of the IL-6 RE-driven gene expression in response to
IL-6. This repression mechanism may be applicable to the E1A repression of
IFN-gamma-induced gene activation.

PMID: 7963529 [PubMed - indexed for MEDLINE]


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