Analysis of premaligment cervical lesions by in-situ hybridization and feulgen-densitometry

Biol. Cell (1991), IN V I T R O ~ O F &HYCOTOXlN : DF,STBOXZN££ ON lnJHAN L E I ~ C C E L L S A N D i l l J ~ L~flPllOC~. CDIER Fran¢o£se u, VAGOPhil£ppe nm eC BUREAU;ean Paul ~m. • Station de Recherches de Pathologie Comparde, 30380 S~-Chr~stol-les-Ales. U~LaboraCo~re de B~olog£e Cellula~ re e¢ ImmunoKdn~¢£que, F a c u l ~ de H~dectne, 30900 HZHES. The interesC £n myco¢oxtns Is rel~Ced to ~helr an~¢v£ral and anC¢¢umoral e f f e c t s . In a previous study, we repotLed thaC a mycotox£n £solaCed from ~he hyphomycete He~ar~lzlum anlsopl£ae (Hetsch) Sorok (Des~ruxin E : DE) ~nhlbx~ed the proltteraCton of mete leukemic c e l l s (P388) " l n v£¢ro" (1). Zn t h i s tnSt£al study, the an~£~umor e f f e c t o f DE was detem£ned ~8 hours a f t e r exposure co the ~ox£n on ~rowth, vlab£1~Cy o~ c e l l s and perturbaC£oas of c e l l cycle us£ng flow cyCometry. The same techntque was used in the presenC s~udy ~o deCerm£ne the ac~£vt¢y o f DE on non sC£mulaCed lymphocy~es and on tumoral human c e l l s . Doses of DE (0.6, 0.06, 0.006 ~8/ml) reduced human leukem:tc c e l l s r a t i o (Number of l / v l n 8 c e l l s 48 hours a f t e r DE exposure I Number of ~v~n~ c e l l s at the be~lnnln8 of ~he exper£ment) by 72%, ~6~ and 28% respec¢£velly. Zn the mouse leukemic c e l l s 6.6, 0.66 and 0.06 MK/ml i~ produced 87%, 82% and 18% decrease in c e l l ra¢£o. Furthermore, an £ncrease in ~he number of c e l l s In Go/1 phase was observed only [n humarl and mouse leukemic c e l l s (cytos~at£c efrec~ ). In nonst£mulated human lymphocytes, the d£Fferen~ doses of DE did not produce any s t s n l r ¢ ; a n t chanBe on c e l l ra¢£o. Thus, our work su~esCs thaC ~he effec~ oC DE was o~serred In p r o l i f e r a t i v e c e l l s only. No e f f e c t was ~ound ~n nomsl c e l l s . (1) ODZER F, VAGO P , , OUZOT J . M . , e t BUREAU J . P . C . R . Acad Sc¢, 3)5, 575-578.

DEVAUCHELLE G. P a r t s , 1987,

F L O W C Y T O M E T R I C EVALUATION OF C E L L P R O L I F E R A T I O N IN H Y P E R P A i t A T l t Y R O I D I S M S VAGOPbi.emel, NGUYEN-TRONG Aaob#2, GODLEWSK! Gui~em?-, M ~ T Y - D O U B L E Cbristiane3, BUREAU Je,n-Paul I

It~lbolwtOl~d 'IbOolo~e,Facult~de Af~JlJe, 34000 tT[oM~IIt'~, 2..¢R,w~ de Ctdm~'e Digem'm e¢ 3Lubomtooe d ;4nxorme Px~o/o~qoe, C I ~ U C~'meeu, JOgOO ,Vimes Flow cymmelry (FCM) is widely used as a rapid method to analyse D N A ¢onumt of a great number of cellson a cellby cell basis.Such an anel~is allowsmewuremant of cellproportionsin the G0/OI, S, and G 2 + M phases of cellcycle, D N A conl~t mud).sisof purulJ~yroid~ recovered by fine needle aspimion in pieces of perlyroldectom~ was pm'ormed for 22 (14 womm and 8 men) pments (15 secondary, hyperperxhyroidism in p~ems with dn'on~c renal f~ure and 7 primary byperpersr~yroidisms). The FCM cell cycle mudvsis has emphasized the s,gmfi~--mt (p
was observed in one preset with a pmlhyroid oxyphiSc ceils l~meery adenomL In two ~ s.~e~ndm'y,hyperpm'alayroidisms, propomon of cells in S and G.~M phases was higher in me primary lesion compared to the mean ~alue of ~eur 8roup (85 • and 8.3 q$).

The study of these cases emphasizes the risk of recurrence of secondary h)'perperslbyroidism with increased proportion of cells in 5_ and G2+M phases with reference to their group mean value,

Therefore, flow cywmemc DNA content unal},'s~s ms~ht assist the surgeon to define the U'emmentof byperpm'aUtycoid pab~Rs,

44a

ANALYSIS OF PREMALIGMENT CERVICAL LESIONS BY IN-SITU HYBRIDIZATION AND FEULGENDENSITOMETRY. SEGERS Pascale. HAESEN Sonja. CASTELAIN Phillipe, VAN HUMMELEN Paul and KIRSCH-VOLDERS Micheline. Laboratorium voor antropo&ene~ica, Vrije Universiteil Brus~ei, Pleinlaan 2, 1050 BRUSSEL, Belgium. Cervical screening is routinely done by morfological examination of epithelium cells on smears, after Papanicolaou staining. The labo- ratory of Antropogcnetics. VUB, Brussels developed recently a new screening method where nuclear DNA content is quantified by means of densitometric image analysis after Feulgen staining on the same slides. It was shown that increasing levels of aneuploidy and polyploidy are correlated with the stage of malignancy. As is the case for many solid tumors (!) chromosome ! seems to play a key role in the development of cervical cancer (2). in an attempt to understand the etiology of these processes and in search of an early marker for premalignant cells in cervical cancer, a nonradioactive in situ hybridization was appliect for the scoring of numerical aberrations of chromosome !. Cervical smears, treated after fixation witk a mucus dissolving agent, underwent hybridization with a biotinylated DNA probe, specific for the centxomeric region of the Iq am1. Visualization is tamed out via FITC-conjugated antibodies, which, after counterstaining of the nuclear DNA with propidium iodide, makes .,,conng of ~pots for chromosoom I possible. Lymfocyte cultures from heahy donors were analysed as controls for the validation of the technique. The preliminary results obtained on smears from controls, and from patients with mild (CIN I) or severe dysplasia (CIN ill) indicate that hypersomy of chrornosooro 1 increases with the malignancy of the lesions. It is postulated that a combined DNA-content/chromosome spot analysis with computer devised image analysis could provide a useful complementar technology for early detection of premaligmem cervical lesions.

(I) ATKIN N.. Cancer Renericscyto.~enencs.21. 279-285.1986. (2) S R E E K A N T A I A H C. C, ncer.62. i3|7-1324. 1988.