Imunohistohemijska analiza i prognostički značaj markera unutrašnjeg puta apoptoze u adenoidnom cističnom karcinomu pljuvačnih žlezda
Immunohistochemical Analysis and Prognostic Value of Markers of the Intrinsic Pathway of Apoptosis in Adenoid Cystic Carcinoma of the Salivary Glands
Author
Dožić, Branko S.Mentor
Boričić, IvanCommittee members
Basta-Jovanović, GordanaTatić, Svetislav

Vukadinović, Miroslav
Metadata
Show full item recordAbstract
Adenoidni cistični karcinom (ACC), je jedan od najčešćih malignih tumora pljuvačnih
žlezda. Učestvuje sa oko 10-15% kod salivarnih neoplazmi. Karakteriše ga spor rast, visoka
incidenca ka invaziji nerva, retko regionalno metastaziranje, česti lokalni recidivi i spor razvoj
progresivnih i relativno indolentnih udaljenih metastaza. Najznačajniji faktori za prognozu ovog
tumora su klinički stadijum i histološki izgled tumora, prisustvo ili odsustvo tumorskog tkiva na
linijama resekcije, invazija nerva.
Apoptoza je regulatorni mehanizam tkivne homeostaze. Poremećaj regulacije apoptoze je
relativno česta karakteristika maligne ćelije. U zavisnosti od stimulansa, putevi aktivacije
apoptoze mogu biti: receptorski (spoljašnji) i mitohondrijski (unutrašnji) put. U unutrašnjem
putu apoptoze značajnu ulogu igraju Apaf-1, inicijatorne i efektorne kaspaze. Apaf-1 je
multidomenski protein, za koji se vezuje citohrom c i prokaspaza 9 što dovodi do formiranja
apoptozoma. Ovaj kompleks proteolitički ak...tivira inicijatornu kaspazu-9. U završnoj fazi
apoptoze, aktivirana kaspaza 9 dovodi do aktivacije efektorne kaspaze 3, čijim ulaskom u jedro
započinje degradaciona faza apoptoze.
Cilj U ovoj studiji cilj je bio ispitati imunohistohemijsku ekspresiju Apaf-1, kaspaze 9 i 3 u
ACC pljuvačnih žlezda, i rezultate korelirati sa kliničko-patološkim parametrima kako bi se
utvrdilo koji je od njih nezavistan prognostički parametar.
Materijal i metode Istraživanje je koncipirano kao studija preseka, koja obuhvata 50 pacijenata
sa ACC pljuvačnih žlezda, muškog i ženskog pola, prosečne starosti 58 godina. U ovoj studiji
koristili smo tehniku tkivnog mikroniza (TMA kalupi). Preseci sa TMA kalupa, debljine 5μm,
bojeni su streptavidin-biotin imunohistohemijskom tehnikom pomoću primarnih antitela
specifičnih za: Apaf-1, kaspazu 9 i kaspazu 3. Obojeni tkivni preseci su analizirani svetlosnim
mikroskopom (Olympus tip BH-2). Na osnovu prikupljenih podataka formirana je baza podataka
u –SPSS 18.0 koja je korišćena za dalju statističku obradu. Statistička analiza podataka
obuhvatala je metode deskriptivne i analitičke (inferencijalne) statistike...
Adenoid cystic carcinoma (ACC), is one of the most common malignant salivary
gland tumors. It participates about 10-15% in salivary neoplasms. It is characterized by a slow
growth, high incidence of the nerve invasion, rare regional metastasis, frequent local recurrence
and slow development of progressive and relatively indolent distant metastases. The most
important factors for the prognosis of this tumor are the clinical stage and histological
appearance of the tumor, presence or absence of tumor tissue on the lines of resection, nerve
invasion.
Apoptosis is the regulatory mechanism of tissue homeostasis. Dysregulation of apoptosis
is a relatively common feature of cancer cells. Depending on the stimuli, activation of apoptotic
pathways may include: receptor (extrinsic) and mitochondrial (intrinsic) pathways. In the
intrinsic apoptotic pathway, Apaf-1, initiator and effector caspases play an important role.
Apaf-1 is a multi domain protein, that binds cytochrome c and procaspase 9, w...hich leads to the
formation of apoptosome. This complex proteolytically activates the initiator caspase-9. In the
final stage of apoptosis the activated caspase 9 leads to activation of the effector caspase 3,
whose entry into the nucleus initiates the degradation phase of apoptosis.
Objective The aim of this study was to examine the immunohistochemical expression of Apaf-
1, caspases 9 and 3 in the ACC of the salivary glands, and to correlate the results with
clinicopathological parameters in order to determine which of them is an independent prognostic
factor.
Materials and Methods The study was designed as a study of sections, which included 50
patients with ACC of the salivary glands, male and female, average age 58 years. In this study,
we used the technique of tissue microarray (TMA moulds). Sections from the TMA mould, 5μm
thick, were stained with the streptavidin-biotin immunohistochemical technique using primary
antibodies specific for: Apaf-1, caspase 9 and caspase 3. Stained tissue sections were analyzed
by the light microscope (Olympus type BH-2). Based on the data collected, the database was
created in -SPSS 18.0, which was used for a further statistical analysis. The statistical data
analysis included methods of descriptive and analytical (inferential) statistics...