Changes of cytoskeletons, extracellular matrices and extracellular matrix receptors in cutaneous squamous cell carcinoma

Posler Presenlations

59

273

276

CHANGES OF CYTOSKELETONS. EXTRACELLULAR MATRICES AND EXTRACELLULAR MATRIX RECEPTORS IN CUTANEOUS SQUAMOUS Mlyoko Kuhn. Shmichl Ohno*. and Toshiaki Saida. CELL CARCINOMA. MediCillC. .Shmshu Univcrsily School of Dcpartmcm 0, Dcrm;mllob~. ““wxany, 0, YallI9”arbi Medical ‘Dcportmcm Anawmy. Malwmtw. Y;im;mn,h\. Japan

Tyr-lie-Gly-Ser.Arg (YIGSR) TERMINAL DIFFERENTIATION

Slgndlcantly mcrca%d amount ot F-acun were dcmonsralcd m all cares of indirect Invrsugaion wnh SCC FITC-phalklidin Wl”l”g. hy rhowcd IhaL Lhe number of high molecular ~mrnun~~llu~lr~sccncc uhniquc wught cyiokcraun (68 kD)-posnwc cells were decrased in ail cases of SCC dctwwd m a poorly d~fferenlialcd SCC. The and vtmtnlin pobnwc cells ucrc cxproaamn t11 lominm ,md lommm receptor wcrc increased m nll casts 01 SCC. accompumcd with mcnlascd amount 01 fihronecun in Lhe surroundmg sLroma. Ordmary c,cc,r
274 KI

HIJMAN

SQUAMOUS

IS A WIDfiPREAD

CELL

Japan

Yamagata.

Kerntins.

epithelial

lnrermediate

construct

keratin

and

differentiation. cultured

solution. ex~ractton.

To

the largest

because

by

dimenstonal

and Northern

solution.

Kl

peptide by a

wlh

pep&de in

an

usually

cultured

step

whjch 1s the pair-kerann

wse

wth

peptide may exsist t&elf

KI. only.

wirh

was

versus by

about

not form

KI

rhe buffer.

prwedure.

was not

68 Kdl Usmg

extraction

whtch

soluhilized

HSCs

and may

68M

8&W%

of

KI /KlO

is

be essenbal

to

dunng terminal

has been

a low

salt

buffer %el

electmphoresir. K

I

peptide

has been done

peptide

may

Moreover. urea

absent aqueous

for keratln

analysis. we demonsrrated

to solubiliry

extraction

basic-types.

as a washing

now. kcratin during

School

consist of 20 proteins(KI-

polyacrylamtde

blot

affinity

respect is

(HSC).

X-100.

CULTURED

Ilniversity

requirements

peptide (KI.

carcinomas

of cultured HSCs. Until X-100

keratinwyres

keratin

cell

and

and functional

saltiTrifon

two

it has developed

/reductam Kl

high

techniques

little unusual

eptdermal the Kl

squamous

fallowed

salt/Triton

wasa

date.

contaimng

proteins.

IN

Katagala. Y utaka Hozumi

Yamagata

acidic-

suprabasal

the structural

immunologral m six kinds

of

filament

families.

maintain human

not

par

Yohtaro

of Dermatology.

KZOI and are dwnded into Iwo acharaclenstic

COMl’ONEN7

CARCINOMAS.

Department

Medicine.

high

-

277 PEPrIDE

KERATIN

and Shigeo Kondo.

in

PROMOTES THE J((eizo Yamamura. tab. of Developmental Biology, National Institute for Dental Research. NIH, Bethesda, MD, USA (KY, MN, HKK) and Dept of Dermatology, Kobe Univ. School of Med., Kobe, Japan (KY, MI) Laminin, a major constituent of basement membrane, has various bioloQical activities with several active sites defined at the synthetic peptide level. Whole laminin is reported to have no effects on ker&nocyte terminal differentiation induced by suspension m methylcellulose, whereas fibronectin is reported to Inhibit the differentiation. In order to reveal the functions of respective active sequences m laminin for keratinocyte terminal differentiation. synthetic peptides containing active sites. Tyr-lie-Gly-Ser-Arg (YIGSR), Ser-lie-Lys-Val-AlaVal (SIKVAV). Arg-Gly-Asp (RGD). and related peptides were prepared and assayed. Neonatal human foreskin keratinocytes were cultured in keratinocyte growth medium with or without these synthetic peptides, and antibodies to involucrin and filaggrin were used as markers of differentiation. Whole laminin showed no effect as already reported. Neither SIKVAV, RGD. nor other related peptides tested in this study had effect on keratinocvte differentiation. Onlv the YIGSR oeobde showed the dlfferentl&on promoting effect at ihe concentratidn bf 30 100 pg/ml. These data demonstrate that a specific site on the laminin may at certain stages regulate keratlnocyte differentiation.

Mofovoshi,

he

the KI

peptide

concentra~wn.

extracted

using

removed Namely.

weaireductanl.

however.

an I-2 M

urea

On

the other

hand. KIO

pepude.

expressed

in HSC%

employed.

So. the

keratin

filament

I” cultured

HSCs.

275 CENTIUWEEPROTEINB IS HICELYEXPRESS&D IN PSOPIATICSKIN. Nanping Rnng,*Auanriang ZhanprYongchro ‘ImnKand Shiyin Li, Department of Dsrmatology,Beijing Medical Unircrsitg &nd ~Departmsnt of Cell Biolory, Beijing Normal Unireraity, Baij ing, P. R.China The Ceattomerc~kinetiehcrr is t spceirliscd sttuctnto locstrd at the primary constriction of sukarpotic rhromosmosn and Innetion@ to attach ehromowmel to mitotie Bpindle.It i8 requited for normal qoremant and diaiunction of chromosomes durina mitosis and urk& aetiologp ad it is ebwretrriscd by blpst-prolifrtition of spidermia. In order to oxplnin the pathologic cell kinetie# in we iarsntigattd CENP-BmItNAexprsuion in normal &ad paoriacia, psoriatic skins. Total RNAwere extracted from hyperprolifsratirs lesions of pwtietics and normel akin of hellthy rolunteers and anrlpned by using BNAblotting hybriditad with L ‘nP-lrb~lltd eDNAprobe. The rrcult dsmonatratrd that CENP-BmftM 118 highly rxwsssed in o8ori&tic akin eomnred mith ncrmxl eontrol.hrsorrt, Gi and Y phr8s btlt,sspeei~lI~,eler~ted during 02 phrse.Thetefare, it ia indicated that elsrrtad CENP-Bsxpreasion ia LP important event in the pathophyaiology of paorinais.

PEPTIDE IN LAMININ OF KERATINOCYTES.

278