Propolis ve etken maddeleri olan kafeik asit fenetil ester (CAPE) ve sinamik asitin, insan T hücreli akut lenfoblastik lösemi hücre dizisi (CCRF-CEM)\'de sitotoksik ve apoptotik etkinliğinin değerlendirilmesi
Propolis bal arıları tarafından kovanda üretilen doğal bir üründür. Çeşitli bitkilerin yaprak, gövde ve tomurcuklarından işçi arılar tarafından toplanıp kovanda biriktirilen propolisin antiseptik, antibakteriyal, antiinflamatuar, immünmodülatör, antioksidan, antimutajenik ve sitotoksik etkileri bilimsel çalışmalarda kanıtlanmıştır. Ancak, akut lenfoblastik lösemi hücre dizisi olan CCRF-CEM ile yapılmış çalışma bulunmamaktadır. Bu çalışmada; propolisin CCRF-CEM hücre dizisinde sitotoksik ve apoptotik özellik gösterip göstermediğinin araştırılması hedeflenmiştir. Sitotoksisitenin değerlendirilmesi için Tripan mavisi yöntemi ve XTT yöntemleri kullanılmış, apoptozisin değerlendirilmesi için ise, apoptozis esnasında açığa çıkan oligonükleotidlerin saptanması esasına dayalı Eliza yöntemi ve apoptotik cisimciklerin flüoresan mikroskopta görüntülenmesini sağlayan Akridin Oranj - Etidyum Bromid boyama tekniği kullanılmıştır. Propolisin sitotoksik etkisinin saptandıktan sonra, propolisin etken maddelerinden Sinamik asit ve CAPE (Kafeik asit fenetil ester)\' in sitotoksik özellikleri aynı yöntemlerle değerlendirilmiştir. Sinamik asit herhangi bir sitotoksik etki göstermez iken CAPE\' nin doz ve zamana bağımlı olarak sitotoksik etki gösterdiği ve IC5o dozunun 1 uM olduğu tespit edilmiştir. Akridin Oranj - Etidyum Bromid boyama yöntemleri ile CAPE\' nin aynı hücre dizisinde apoptozise neden olduğu bulunmuştur. CAPE, normal hücreler üzerinde düşük toksisiteye sahip, yeni antimetastatik ve antianjiyojenik ajan olarak değer taşıdığından bu sonuçlar akut lenfoblastik lösemi tedavisinde propolis ve özellikle CAPE\' nin kullanılabilmesine yönelik yeni bir pencere açmıştır.
The evaluation of cyctotoxic and apoptotic effect of propolis and its extracts caffeic acid phenethyl ester and cinnamic acid in human acute T cell lymphoblastic leukemia cell line (CCRF-CEM)
Propolis is one of the nature- based products, which is produced by honeybees in hives. It is collected by honey bees from leaves, trunk and bud of several different plants. Propolis has antiseptic, antibacterial, anti-inflammatory, immu-nomodulatory, antioxidant, antibiotic, antimutagenic and cytotoxic effects which are proven in scientific studies. There has been no study in CCRF-CEM cell line with propolis and its components in the literature. The aim of this study is to investigate the cytotoxic and apoptotic effects of propolis and its components in CCRF-CEM cell line. Trypan blue dye exclusion test and XTT methods are used to evaluate the cytotoxicity and ELISA based on detection of oligonucleotids which can be seen during apoptosis. Acridine orange / Ethidium Bromide dye technique based on detection of apoptotic bodies with Fluorescent microscope were used to evaluate apoptosis. After detection of cyto-toxic effect of propolis, the cytotoxic effect of. Cinnamic acid and CAPE which are the effective components of propolis, were evaluated with the same methods. While cinnamic acid didn\'t have any cytotoxic effect, CAPE had dose and time dependent cytotoxicity and had 1 /jM IC50 in CCRF-CEM cell line. ELISA and Acridine Orange / Ethidium Bromide dye methods revealed that CAPE induced apoptosis in the same cell line. Since CAPE is a new chemotherapeutic and antitumoral agent with very less toxic effects on normal tissues, these results opened a new horizon for use of propolis in the treatment of acute lymphoblastic leukemia.
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