School of Dental Medicine, University of Zagreb, Croatian Dental Society - Croatian Medical Association
Abstract
Nearomatski visokosteranski naftalan (NAVS) posebna je frakcija hrvatske nafte, iznimno bogata steranima, iz koje je odstranjen aromatski sastav. Steranima se pripisuje bioaktivnost slična steroidnim hormonima, modulatorima tumorskoga rasta. Potaknuti rezultatima prijašnjih in vitro i in vivo studija o inhibitornom efektu NAVS-a na rast planocelularnog karcinoma, proveli smo studiju o njegovu učinku na oralni planocelularni karcinom (OPCC) u miševa.
Ciljevi: 1) ispitati valjanost jednostavnog eksperimentalnog modela OPCC-a; 2) ispitati mogući antiproliferativni učinak NAVS-a na postavljenome modelu praćenjem tumorskoga rasta; 3) ispitati antineoangiogeni učinak NAVS-a kao objašnjenje mogućeg antiproliferativnog učinka, te za tu prigodu procijeniti mogućnost križne reaktivnosti anti-humanih imunohistokemijskih markera s mišjim tkivom. Suspenzija od 100 μl s 105 SCC VII stanica inokulirana je intraoralnim putem pod bukalnu sluznicu u 48 singeničnih C3H miševa. Sedam dana nakon inokulacije životinje su podijeljene u 6 jednakih skupina, te se miševima, ovisno o skupini, intratumorski injiciralo po 100 μl sljedećih tvari: parafinsko ulje (PO) kao negativna kontrola, NAVS (u jednoj skupini 7 dana, a u drugoj skupini 14 dana nakon inokulacije tumora), 1, 25 dihidroksiergotamin (1,25-D3) kao pozitivna kontrola, te kombinacije NAVS s 1,25-D3 i PO s 1,25-D3. Rast tumora praćen je tjednim mjerenjem s pomoću kalipera. Životinje su žrtvovane 1., 2., 3. i 4. tjedna nakon aplikacije ispitivanoga sredstva. Histološki pripravci eksplantiranih tumora bojeni su hematoksilin-eozinom te imunohistokemijski s anti-CD34 protutijelima radi procjene tumorske neoangiogeneze. U usporedbi s PO skupinom, tumorski rast i angiogeneza bili su sniženi u 1,25-D3 i NAVS skupinama. NAVS vjerojatno smanjuje rast OPCC-a inhibicijom vaskularne proliferacije potrebne za tumorski rast.Nonaromatic naphthalane (NAVS) is a specific fraction of Croatian oil, extremely rich in steranes from which the aromatic system is removed. Steranes are attributed with bioactivity similar to steroid hormones, modulators of tumour growth. Encouraged by the results of earlier in vitro and in vivo studies on the inhibitory effect of NAVS on the growth of planocellular carcinoma, we carried out a study on its effect on oral planocellular carcinoma (OPCC) in mice.
Aims: 1) To test the validity of the simple experimental model OPCC, 2) to test possible antiproliferative effect of NAVS on the above model by monitoring tumour growth, 3) to test the antineoangiogenic effect of NAVS to explain the possible antiproliferative effect, and to estimate the possibility of crisis reactivity of anti-human immunohistochemical markers with mice tissue. A suspension of 100 μl s 105 SCC VII cell was inoculated intraorally under the buccal mucous membrane in 48 syngeneic C3H mice. Seven days after inoculation the animals were divided in six equal groups and the mice, depending on the group, were intratumorously injected with 100 μl of the following substances: paraffin oil (PO) as a negative control, NAVS (in one group 7 days, and in the second group 14 days, after inoculation of the tumour), 1.25 dihydroxyergotamine (1.25-D3) as a positive control, and a combination of NAVS with 1.25 D3 and PO with 1.25- D3. Tumour growth was monitored weekly by measuring with callipers. The animals were sacrificed 1, 2, 3 and 4 week after application of the tested substance. Histological specimens of explanted tumours were stained with hematoxylin-eozine, and immunohistochemically with anti-CD34 antibodies for estimation of tumour neoangiogenesis. Compared with the PO group, tumour growth and angiogenesis were decreased in the 1.25-D3 and NAVS groups. NAVS probably reduced growth by OPCC inhibition of vascular proliferation, needed for tumour growth
