Expression of thrombospondin in BPH, pin, and prostate cancer

230

229 EXPRESSION PATTERN OF THE ADAMTS HUMAN PROSTATE CANCER CELL LINES Chandrasekharan ‘University Xlniversity Kingdom

PROTEINASES

IN

AT

NUCLEOTIDE

S.‘. Hamdy F.C.‘, Holen I.‘, Cross N.‘, Buttle D.J.’

of Sheffield. Dept. of Urology, of Sheffield. Division of Genomic

INSTABILITY

Sheffield, Medicine,

SELECTED

REPEATS

MICROSATELLITE

IN PROSTATE

TETRA-

CANCER

Arrou~i A.R.‘, Catto J.W.‘. Rchman I.l_ Feeley K.‘, Meuth M.‘. Hamdy F.C.’

United Kingdom, Shefftield, United

‘Royal

Hallamshire

Kingdom.

INTRODUCTION & OBJECTIVES: Cancer invasion requires the breakdown of basement membranes at an early stage. as well as angiogenesis. which is csscntial for tumour growth and haematogenous spread. The ADAMTS enzymes, named because of their structure and function (A Disintegrin And Metalloproteinase with ThromboSpondin motifs) modulate tumour growth, inhibit angiogenesis and are thought to bc involved in metastases formation. Our objective was to study the differential expression of a selected subgroup ofthese enzymes in human prostate cancer cell lines.

RESULTS: ADAMTS -5. 9 and 15 are only expressed by the hormone independent prostate cancer cell-lines PC-3 and DUl45, while ADAMTS-I and 4 arc expressed by both as well as the androgen-sensitive LNCaP cells. We did not find ADAMTS-8 cxprcssion in any of these cell lines. CONCLUSIONS: To date, there have been no prior published studies investigating ADAMTS enzyme expression in prostate cancer. The differential expression of these enzymes in vitro suggests that whilst angiogenesis inhibiting ADAMTS-1 and 4 may limit aggressive behaviour of the cells a well know feature of LNCaP cells, over-expression of ADAMTS -5, 9 and 15 mzay contribute to the hormone resistant phenotype. Further studies arc in progr-css quantifymg enryme levels. and modulating their expression in vitro. The findings and their implications wtll be discussed.

Academic

of Shcfflcld.

Urology

lnatltute

Unit,

for Cancer

Shcftield, Studies.

INTRODUCTlOlr; role

& OBJECTIVES:

in human

repetitive

carcinogenesis.

nucleotides

Microsatcllitc

that

instability

Cicnetlc instability

Microsatcllites can

be used

are

to study

(MSI), as charactcrised

regions

genomic

by hereditary

colon cancer. is caused by a lhilure of the mismatch form of MSI has recently

plays an important

ubiquitous

D~rcvl N.‘, Bjartell A.B. Abrahamsson P.’

AND SEROTONIN AND IN PROSTATE

, Nilsson F. . Hansson _I.‘. (iadalcanu L’.‘. Hamdy 1:.

‘Malmii University Ilospital. Urology, Malmd. Swcdcn, Malmd Univcrslty Hospitnl. Pathology. Malmii. Sweden. University of Neucaslle. Clrology, Sheffield. tinitctl Kingdom INTRODUCTION & OBJECTIVES: Prostate cancer (PC‘s) cells often develop androgen-independent growth after androgen suppression therapy. The number of serotonin (5-HT) releasing ncuroendocrine (NE) cells is greater III prostate carcinoma compared to normal prostatic tissue, and correlates with tumour progression, loss of androgen dependence and poor prognoals. 5-HT is a biogenic amine I-elated to prostate growth and development. The objective of this study was to investigate the presence and significance of S-HT receptor (5.HTR) subtype I HI benign and malignant prostatic tissue, metastascs and PCs cell lines. MATERIAL & METHODS: Prostate tissue specimens and metastases were obtained from 35 patients (mean age 74 years) undergoing TIJRP, lamincctomy or pelvic lymph node dIssectIon, respectively. Cultured tumour cells (LNCaP, PC-3 and DUl45), benign prostatic stromal cells (hPCPs) and PC-3 cells implanted in athymic nude mice were also included in the study. Western blotting and i~nmunohistochcmistry were performed to detect 5-IITR IA, IB and ID protein expression and RT-PCR was utdized to evaluate ImRNA expression. The effect of 5HTlA antagonists on PCs cell lines was studied by BrdU ELlSAproliferation aasaya. RESULTS: lmmunohistochemistry rev&d subtype 5-HTRIA and IB to bc localized in the cytoplasm of carcinomatous cells but only in few BPH cells, whereas 5.HTRID was confined to vascular endothelial cells. An increase of number and unmunstaining reactivity was noted in tumours of high Gleason grade (4-5). We demonstrated the expression of 5-HTRI A in vitro in androgen-independent PC-3 and DU145 cell lines and in vivo in PC-3 cells implanted in nude mice. In contrast, barely detectable levels of 5-HTRIA were found in the androgen-sensitive LNCaP cell line and stromal hPCPs cells. Furthermore, 5.HTRIA mRNA was demonstrated in PC-3 and LNCaP PCs cell lines by RT-PCR. Using BrdU ELISA proliferation assay, 5HTRIA (NAN-190) showed growth inhibition of PC-3, DUl45 and LNCaP PCs cell lines, but not in benign prostatic stromal cells.

of

instability, non-polyposis

repair system. A genetically

been described

m which

alteration5 are present at sclccted trt- and tctranuclcotidc

microsatclllte

repeats (LMAST). We

evaluated this form of MS1 in prostate cancer. \%ATERIAL &tamed

& METHODS:

from 20 specimens

DNA extraction tetra-nucleotide

Iumoural

and normal

of radical prostatectomy

and PCR wnh tluorcacently repeats, microsatcllitc

proslale

wcrc

by Iilicl-odiascctlon.

adlaccnt

After

labelled priniers

analysis was pcrformcd.

for 3 different The criteria used

for F.MAST(+) was the presence of new or altered band in the tumoural DNA. RESULTS:

All tumours were successfully

tetra-nuclcotide

studied. Instability

was seen at the

loci in 5/2O (25%) tumours. One tumour showed instability

at

more than one loci (11~2). CONCLUSIONS:

Microsatellite

repcats studied occurs in a quarter role in the parhogenesia

instability

at the >electcd

of prostatecancer,.

terra-nuclcotide

It may pla) an important

ol‘a subset of prostate cancer.

232

231 EXPRESSION OF SEROTONIN RECEPTORS ACTIVITY IN HUMAN PROSTATE CANCER CANCER CEI.1. LINES

United Sheffield,

United Kingdom

distinct .MATERIAL & METHODS: Three prostate cancer cell lmes were used, nndrogen independent PC-3 and DlJ145 lines. and the androgen dependent LNCaP line. Cells were grown to confluence using standard medium, then RNA extracted and cxpresston studied by RT-PC‘R using custom deslgned primers for ADAMTS-I. 4, 5, 8. 9 and 15. PCR products were purified and sequenced to ensure that the appropriate product was Idcntltird.

IIospital,

Ylniversity

EXPRESSION

OF THROMBOSPONDIN

IN BPH, PIN, AND PROSTATE

CANCER L’Ylbo H.. Damber J. Inst. of Surgical Sclencea, Urology. Gdtcborg, INTRODUCTION

& OBJECTIVES:

be of great importance mvolved

Swcdcn

rumour

angiopcnesls

IS considered

lo

for tumour growth. Several factors are identified as bemg

in the process.

Thrombospondin

(TSP) has shown to be a negative

I-cgulator of angiogenesis

in many human tllmourb. The aim ol’thia study was to

evaluate

of thrombospondin

the expression

(BPII) and prostate \IATERlAL u,lng

& METHODS:

Immunohlstochemistry

ti\\uc spccimcnc.

m benign

prostatic

hyperplasla

cancer.

The cxprcssion

was a\~~ed

of TSP (TSP.I

In this material there \\ere 32 patient\

and TSP-2) by

and prostate wccr

1117.: pro\tatr

\vith RPH. 7 patients

with PIN and 34 patrrnrs with prostate cancer. RESULTS:

The result from the immunohistochemistry

showjed that 26 of 32

(81%) patients with BPH were positive for TSP-I. All patlents with PIN were positive for TSP-

I. Two out of 34 (6%) cancer patients were positive for TSP-I.

The cancer patients were divided into two groups depending resulted in that 4,50/o were TSP.1 positive in Gleason

on Gleason score

S-7 group and X,3% in

Gleason group 8-10. All 73 patients were negative for TSP-2. CONCLUSIONS:

Thrombospondin,

a known inhibitor angiogenesis.

is almost

CONCLUSIONS: This is the first report that demonstrates an upregulation of 5-H-1.K subtype IA and 1B in PCs, especially in high-grade turnours. Moreover, antagonists to 5.HTRlA inhibit proliferation of PCs cells. We propose an important role of 5-HT

totally absent in prostate cancer while BPH expresses TSP m nearly all samples.

in tamour progression,

with cancer decelopmcnt

especially

in the androgen-independent

European Urology Supplements

state of the disease.

2 (2003) No. 1, pp. 60

This linding is suggestive

for a role of TSP in the angiogenic in the prostate.

shift associated