Ecem KAYA SEZGİNER, Nuri ÖZMEN, Filiz BAKAR ATEŞ, Emin Emre KURT

Kolşisin’in MCF-7 insan meme adenokarsinoma hücrelerinde hücre döngüsü tutulumu ve MMP-2 mRNA ifade seviyesi üzerine etkisi

Amaç: Kolşisin, trisiklik alkaloid bir ilaç olup klinikte anti-enflamatuvar etkisinden dolayı yaygın olarak kullanılmaktadır. Kolşisin uygun konsantrasyonlarda oral yolla kullanılmakta iken yüksek konsantrasyonlarda toksik etki gösterdiği tespit edilmiştir. Bu nedenle, kanser araştırmalarında düşük doz kolşisinin etki mekanizmaları üzerine çalışmalar yapılmaktadır. Matriks metalloproteinaz-2 (MMP-2), matriks metalloproteinaz enzim ailesinin bir üyesi olup kanserde ifade seviyesinin arttığı ve ekstrasellüler matriksi yıkıma uğratarak kanser hücrelerinin metastazına neden olduğu bildirilmiştir. MMP’lerin bu önemli etkisi nedeni ile kanser hastalığında metastazın önlenmesi amacıyla çeşitli bileşiklerin MMP üzerine inhibitör etkisinin değerlendirilmesi kanser araştırmalarında önemli yer tutmaktadır. Bu çalışmada, kolşisinin, MCF-7 insan meme adenokarsinoma hücresinde hücre döngüsü tutulumu ve metastazda önemli rol oynadığı bilinen MMP-2 protein ifadesi üzerine etkisinin araştırılması amaçlanmıştır. Yöntem: Çalışmada, MCF-7 insan meme adenokarsinoma hücreleri kullanılmıştır (ATCC, HTB22). Hücrelere 0.1, 10 ve 100 μg/ml konsantrasyonlarda kolşisin uygulanmış ve hücre canlılığı analizi MTT testi ile yapılmıştır. Kolşisin’in hücre döngüsü tutulumu üzerine etkisi analiz edilmiş ve G0/G1, S ve G2/M döngüsündeki hücre populasyonları yüzde (%) olarak ifade edilmiştir. Kolşisin’in MMP-2 mRNA ifade seviyesi üzerine etkisi ise gerçek zamanlı PCR (qRT-PCR) yöntemi ile tayin edilmiştir. Bulgular: 10 ve 100 μg/ml konsantrasyonlarda Kolşisin MCF-7 meme kanser hücresinin canlılığını istatistiksel olarak anlamlı derecede inhibe etmiştir. Kolşisin’in tüm konsantrasyonlarda hücre döngüsünü G2/M fazında inhibe ettiği ve MMP-2 geninin ifade seviyesini anlamlı olarak azalttığı saptanmıştır (p

Effects of colchicine on cell cycle arrest and MMP-2 mRNA expression in MCF-7 breast adenocarcinoma cells

Objective: Colchicine is a tricyclic alkaloid drugand it’s been clinically used for a long time because ofits anti-inflammatory effects. While colchicine has beensafely used in oral applications, it’s been demonstratedthat colchicine is toxic at higher concentrations. So,the mechanism of effects of low dose colchicine hasbeen studied in cancer related researches. Matrixmetalloproteinase-2 (MMP-2) is a member of matrixmetalloproteinase enzyme family and it’s beendemonstrated that MMP-2 expression is increased incancer and it causes a metastasis of cancer cells throughdegradation of extracellular matrix. Because of thisimportant effect of MMPs, the studies related to developMMP inhibitor compounds have great importance incancer investigations. At present study, it’s aimed toinvestigate the effects of colchicine on cell cycle arrestand MMP-2 mRNA expression in MCF-7 human breastcancer cells.Methods: In the study, MCF-7 human breastadenocarcinoma cells were purchased from ATCC.Cells were treated with 0.1, 10 ve 100 μg/mlcolchicine and cell viability was determined via MTTassay. The effect of colchicine on cell cycle arrest wasdetermined by Muse Cell Analyzer and the percent ofcell populations at G0/G1, S and G2/M cycles wereidentified. The effect of colchicine on MMP-2 mRNAexpression has been performed by real-time PCR (qRTPCR) analysis.Results: Colchicine has significantly inhibitedcell viability at 10 and 100 μg/ml concentrations. It’sbeen also demonstrated that colchicine has induceda cell cycle arrest at G2/M phase and downregulatedMMP-2 mRNA expression of MCF-7 cells in all treatedconcentration (p

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