Imunohistohemijska analiza i prognostički značaj markera unutrašnjeg puta apoptoze u adenoidnom cističnom karcinomu pljuvačnih žlezda

Abstract

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 aktivira 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, which 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...